The physical factors affecting the production of protease from bacillus cereus was investigated. Agro
industrial waste product groundnut shells were used as substrates in solid-state fermentation for protease enzyme
production. The response surface methodology (RSM) was used to optimize protease production by implementing the
central-composite design (CCD). The physiological fermentation factors such as pH (8.0), temperature (43°C),
fermentation time (26 hrs), inoculum level (3.2 ml) and substrate concentration (9.6g) were optimized by statistical
analysis using response surface methodology. The maximum yield of protease production was 76.75U/gds. This was
evidenced by the higher value of coefficient of determination (R2= 0.9994).
This document describes a new method for immobilizing DNA, proteins, and enzymes on poly(dimethylsiloxane) (PDMS) biochips. Specifically:
1. Biologically active compounds like DNA, antibodies, and enzymes are attached to micrometer-sized beads.
2. The beads are then spotted onto a poly(vinyl chloride) surface and dried, forming an array.
3. PDMS is molded over the bead array, entrapping the beads in the polymer.
4. This allows the creation of either low-density arrays or integrated fluidic chips with the sensing elements directly incorporated into microchannels.
5. The method was shown to successfully create arrays for
- The document summarizes the MSc thesis of Mauro José Castanho Claudino which aimed to screen and characterize different immobilization methods for whole-cell steroid bioconversion using Mycobacterium sp. NRRL B-3805.
- Silicone was found to be the most suitable immobilization support, providing efficient cell adsorption, good thermal stability, and ability to catalyze the biotransformation of β-sitosterol to 4-androstene-3,17-dione over multiple batches. Kinetic studies showed Michaelis-Menten behavior and storage stability could be modeled using bi-exponential equations.
- The work demonstrated the feasibility of using small-scale bioreactors to
This document summarizes a study investigating in vitro dissolution differences between BCS class 1 drugs in South American markets. The study compared products in three countries to US reference listed drugs (RLDs) for zidovudine, amoxicillin, and metronidazole. The study protocol involved testing 12 units of each product in pH 1.2, 4.5 and 6.8 buffers to generate dissolution profiles and calculate similarity factors (F2) between products and RLDs. The goal was to determine if biowaivers could be granted for generics with similar dissolution profiles to avoid in vivo bioequivalence testing.
This document summarizes research on the synthesis of cerium oxide nanoparticles (CeO2-NPs) using honey. Key points:
- Researchers developed a green chemistry method to synthesize CeO2-NPs using honey through a sol-gel process. This provides an environmentally friendly alternative to conventional synthesis methods.
- Characterization techniques confirmed the successful production of spherical CeO2-NPs around 23 nm in size with a fluorite cubic structure.
- In vitro cytotoxicity tests on neuro cells showed the CeO2-NPs were not toxic at concentrations below 25 mg/mL, indicating potential for biomedical applications.
- Using honey proved comparable to other methods for synthesizing
Determination of Immobilization Process Parameters of Corynebacterium glutami...IJERA Editor
The parameters of the immobilized process of Corynebacterium glutamicum on kappa carrageenan were identified by
Plackett-Burman matrix, and the experiments were designed by response surface methodology having the central
composite designs (RSM-CCD). The maximum yield of cell immobilization on kappa carrageenan carrier reached at
78% ± 2%. Optimal parameters were 3 grams kappa carrageenan per 100 militters sterile water and 58.58 million
cfu/mL, forming gels at 100C for 25 minutes and the speed when soaking particles of 150 rpm for 120 minutes in 0.58
M potassium chlorua solvent. The immobile finished products are applied in L-lysine production, their reusing ability
is 3 times and the total yield of L-lysine was accumulated 93 g/L in medium during 96 fermented hours. The L-lysine
productivity of the batch fermentation was 0.969 g.L-1
.h-1
. And the set-up storage conditions are the mixed solvent of
CaCl2 0.5% (w/v) and KCl 0.5% (w/v); pH is 7.0 in 40C. After 60 storage days, the survival cell rate was remained
51%.
The microbiology of the winemaking process, which includes inoculated strains
of the yeast Saccharomyces cerevisiae and the lactic acid bacterium, Oenococcus
oeni, is critical to process efficiency and wine quality. In each case these organisms
are required to complete a core conversion (sugar to ethanol or lactate to malate,
respectively) as well as make desirable sensory contributions. These activities
typically occur under extreme conditions which may include high sugar (osmolarity)
and ethanol content and low pH, temperature and nutrient availability. We have used
mutant screening strategies and functional genomic approaches to identify the basis
of superior yeast performance in the face of these challenges. In addition we have
use adaptive evolution to yield yeast with enhance fermentation reliability based on
increase nitrogen efficiency, fructophilicity or general robustness. In parallel work,
we have isolated and heterologously expressed genes from O. oeni which encode
esterases or glucosidases. Characterisation of these gene products has provided
insights into their roles within the cell as well as potential contribution to wine.
Biolostic transformation of a procaryote, bacillus megateriumCAS0609
This document describes a new method for transforming the bacterium Bacillus megaterium using biolistic transformation. Key findings include:
1) Plasmid DNA was coated onto tungsten microprojectiles and accelerated into B. megaterium cells using a helium-driven biolistic device.
2) Over 104 transformants per treated plate were obtained after optimizing biological and physical parameters of the biolistic process.
3) All strains of B. megaterium tested were successfully transformed, though efficiency varied between strains. This is the first report of biolistic transformation of a prokaryote.
Towards understanding genetic basis of chapatti (Indian flat bread) making qu...CIMMYT
The document discusses a study on understanding the genetic basis of chapatti (Indian flat bread) making quality. Researchers analyzed samples from different wheat cultivars and identified genes associated with chapatti quality scores. Gene expression profiling identified differentially expressed genes between good and poor quality cultivars at different developmental stages. Upregulated genes in good quality cultivars were associated with storage proteins and nutrient reservoir activity, while downregulated genes were related to catalytic activity and starch biosynthesis.
This document describes a new method for immobilizing DNA, proteins, and enzymes on poly(dimethylsiloxane) (PDMS) biochips. Specifically:
1. Biologically active compounds like DNA, antibodies, and enzymes are attached to micrometer-sized beads.
2. The beads are then spotted onto a poly(vinyl chloride) surface and dried, forming an array.
3. PDMS is molded over the bead array, entrapping the beads in the polymer.
4. This allows the creation of either low-density arrays or integrated fluidic chips with the sensing elements directly incorporated into microchannels.
5. The method was shown to successfully create arrays for
- The document summarizes the MSc thesis of Mauro José Castanho Claudino which aimed to screen and characterize different immobilization methods for whole-cell steroid bioconversion using Mycobacterium sp. NRRL B-3805.
- Silicone was found to be the most suitable immobilization support, providing efficient cell adsorption, good thermal stability, and ability to catalyze the biotransformation of β-sitosterol to 4-androstene-3,17-dione over multiple batches. Kinetic studies showed Michaelis-Menten behavior and storage stability could be modeled using bi-exponential equations.
- The work demonstrated the feasibility of using small-scale bioreactors to
This document summarizes a study investigating in vitro dissolution differences between BCS class 1 drugs in South American markets. The study compared products in three countries to US reference listed drugs (RLDs) for zidovudine, amoxicillin, and metronidazole. The study protocol involved testing 12 units of each product in pH 1.2, 4.5 and 6.8 buffers to generate dissolution profiles and calculate similarity factors (F2) between products and RLDs. The goal was to determine if biowaivers could be granted for generics with similar dissolution profiles to avoid in vivo bioequivalence testing.
This document summarizes research on the synthesis of cerium oxide nanoparticles (CeO2-NPs) using honey. Key points:
- Researchers developed a green chemistry method to synthesize CeO2-NPs using honey through a sol-gel process. This provides an environmentally friendly alternative to conventional synthesis methods.
- Characterization techniques confirmed the successful production of spherical CeO2-NPs around 23 nm in size with a fluorite cubic structure.
- In vitro cytotoxicity tests on neuro cells showed the CeO2-NPs were not toxic at concentrations below 25 mg/mL, indicating potential for biomedical applications.
- Using honey proved comparable to other methods for synthesizing
Determination of Immobilization Process Parameters of Corynebacterium glutami...IJERA Editor
The parameters of the immobilized process of Corynebacterium glutamicum on kappa carrageenan were identified by
Plackett-Burman matrix, and the experiments were designed by response surface methodology having the central
composite designs (RSM-CCD). The maximum yield of cell immobilization on kappa carrageenan carrier reached at
78% ± 2%. Optimal parameters were 3 grams kappa carrageenan per 100 militters sterile water and 58.58 million
cfu/mL, forming gels at 100C for 25 minutes and the speed when soaking particles of 150 rpm for 120 minutes in 0.58
M potassium chlorua solvent. The immobile finished products are applied in L-lysine production, their reusing ability
is 3 times and the total yield of L-lysine was accumulated 93 g/L in medium during 96 fermented hours. The L-lysine
productivity of the batch fermentation was 0.969 g.L-1
.h-1
. And the set-up storage conditions are the mixed solvent of
CaCl2 0.5% (w/v) and KCl 0.5% (w/v); pH is 7.0 in 40C. After 60 storage days, the survival cell rate was remained
51%.
The microbiology of the winemaking process, which includes inoculated strains
of the yeast Saccharomyces cerevisiae and the lactic acid bacterium, Oenococcus
oeni, is critical to process efficiency and wine quality. In each case these organisms
are required to complete a core conversion (sugar to ethanol or lactate to malate,
respectively) as well as make desirable sensory contributions. These activities
typically occur under extreme conditions which may include high sugar (osmolarity)
and ethanol content and low pH, temperature and nutrient availability. We have used
mutant screening strategies and functional genomic approaches to identify the basis
of superior yeast performance in the face of these challenges. In addition we have
use adaptive evolution to yield yeast with enhance fermentation reliability based on
increase nitrogen efficiency, fructophilicity or general robustness. In parallel work,
we have isolated and heterologously expressed genes from O. oeni which encode
esterases or glucosidases. Characterisation of these gene products has provided
insights into their roles within the cell as well as potential contribution to wine.
Biolostic transformation of a procaryote, bacillus megateriumCAS0609
This document describes a new method for transforming the bacterium Bacillus megaterium using biolistic transformation. Key findings include:
1) Plasmid DNA was coated onto tungsten microprojectiles and accelerated into B. megaterium cells using a helium-driven biolistic device.
2) Over 104 transformants per treated plate were obtained after optimizing biological and physical parameters of the biolistic process.
3) All strains of B. megaterium tested were successfully transformed, though efficiency varied between strains. This is the first report of biolistic transformation of a prokaryote.
Towards understanding genetic basis of chapatti (Indian flat bread) making qu...CIMMYT
The document discusses a study on understanding the genetic basis of chapatti (Indian flat bread) making quality. Researchers analyzed samples from different wheat cultivars and identified genes associated with chapatti quality scores. Gene expression profiling identified differentially expressed genes between good and poor quality cultivars at different developmental stages. Upregulated genes in good quality cultivars were associated with storage proteins and nutrient reservoir activity, while downregulated genes were related to catalytic activity and starch biosynthesis.
Estimation Of The Nucleation Kinetics For The Antisolvent Crystallization Of ...cliff57
This document discusses estimating the nucleation kinetics for the anti-solvent crystallization of paracetamol in methanol-water solutions. Specifically:
- Laser back-scattering (focused beam reflectance measurement or FBRM) was used to detect nucleation events during anti-solvent crystallization experiments with paracetamol.
- Theoretical models were applied to analyze induction time and metastable zone width (MSZW) data from these experiments and estimate nucleation kinetics parameters.
- Solvent composition, in terms of the methanol-water ratio, was found to significantly impact measured induction times and MSZWs. Estimated nucleation rates also decreased with more dynamic solvent compositions.
-
This document describes a study on genetic diversity in soybean (Glycine max) using RAPD marker. Six soybean genotypes were collected and their DNA was extracted using the CTAB method. The DNA was quantified and found to range from 158-502 ng/μl. The objectives were to analyze genetic diversity among the genotypes using RAPD marker. The methodology involved DNA extraction, quantification, PCR amplification with RAPD primers, and resolving the amplified products through gel electrophoresis. The results showed good quality DNA was extracted. The study provides information on genetic variation that can aid in soybean breeding programs.
Determination Of The Crystal Growth Rate Of Paracetamol As A Function Of Solv...cliff57
1) The document discusses a study that determined the crystal growth rate of paracetamol as a function of solvent composition using antisolvent crystallization in methanol-water mixtures.
2) Crystal growth rate was found to decrease with increasing water content up to 68% water, at which point the growth rate increased.
3) The study introduced a method to link solvent composition to growth mechanism and rates. It was postulated that solubility gradient, viscosity, selective adsorption, and surface roughening affect growth rates with solvent composition.
1) The document describes a rapid method for extracting genomic DNA from filamentous fungi that involves bead beating to disrupt fungal cell walls followed by phenol-chloroform extraction and isopropanol precipitation.
2) The method yields 60-230 μg of high quality DNA per 200 mg of fungal mass within 2.5 hours without enzymatic digestion.
3) The extracted DNA was suitable for downstream PCR applications like gene amplification and RAPD analysis, demonstrating its utility for high-throughput fungal identification.
Detection of Genetically Modified Soybean in Crude Soybean Oil.PDFGordana Zdjelar
This document describes a study that aimed to detect and quantify the presence of Roundup Ready genetically modified soybean in crude soybean oil extracted from soybean seed with varying percentages of GM content. Two DNA extraction methods were compared: CTAB and a commercial DNeasy Plant Mini Kit. Real-time PCR was used to amplify DNA targets from the extracted samples to detect and quantify the GM content. The results showed that the commercial kit was superior to CTAB extraction for recovering DNA from crude oil samples. Both qualitative and quantitative PCR analysis demonstrated it is possible to monitor the GM content in crude oil derived from soybean seed with varying GM percentages.
The document describes the identification, cloning, sequencing, and characterization of the a-L-arabinofuranosidase B (abfB) gene from the phytopathogenic fungus Fusarium oxysporum f. sp. dianthi (Fod). The gene was identified using random amplified polymorphic DNA and encodes a protein of 499 amino acids. The recombinant protein expressed in E. coli had arabinofuranosidase activity and optimal activity at pH 4.0 and 50°C. Reverse transcription PCR showed that the abfB gene is actively transcribed in carnation plants infected with Fod, suggesting it plays a role in the fungus's pathogenicity.
This study demonstrates for the first time the production of recombinant human growth hormone (rhGH) from Bacillus subtilis. A hybrid gene containing the signal sequence of the B. licheniformis serine alkaline protease gene and the cDNA encoding hGH was cloned into the pMK4 plasmid and expressed under the deg promoter in B. subtilis. The rhGH produced was secreted and confirmed to have the correct mature hGH sequence. The highest rhGH concentration of 70 mg/L was obtained at 32 hours with a yield of 9 g/kg. Fermentation characteristics differed between B. subtilis containing the hybrid gene versus the plasmid alone. This expression system could be applicable for heterologous protein production from Bac
Synthesis, Molecular Docking and Antimicrobial Evaluation of New Tetrahydrobe...ijtsrd
A series of novel derivatives of Tetrahydrobenzothienopyrimidine hydrazone were synthesized and product structure was elucidated by 1NMR, C13NMR and mass spectroscopy. The synthesized compounds were evaluated against fungal and bacterial strains. The synthesized compounds showed significant antibacterial activity against Staphylococcus aureus MTCC 96, Staphylococcus pyrogenes MTCC 442, and Escherichia coli MTCC 443, Pseudomonas aeruginosa MTCC 1688 and against fungal strains Candida albicans MTCC 227, Aspergillus niger MTCC 282, Aspergillus clavatus MTCC 1323. Some derivatives showed promising result against gram positive, gram negative bacterial and fungal strains than standard drug ampicillin and grieseofulvin. In- silico molecular docking studies of the synthesized compounds was done by using GRIP batch docking method of Vlife MDS 3.0 software to study their observed activity which showed a significant correlation between the binding score and biological activity for synthesized compounds. Neetu Chopra | Kiranpreet kaur | Sanjeev Kumar "Synthesis, Molecular Docking and Antimicrobial Evaluation of New Tetrahydrobenzothienopyrimidine Derivatives" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-2 | Issue-6 , October 2018, URL: http://www.ijtsrd.com/papers/ijtsrd18756.pdf
1) The document describes an automated process for in vitro selection that was developed to generate nucleic acid aptamers faster than the traditional manual selection process.
2) An augmented Beckman Biomek 2000 pipetting robot was programmed to automate the major steps of in vitro selection, including preparing and purifying RNA, filtering RNA-target complexes, and amplifying selected sequences, in order to reduce the time needed to select aptamers from weeks or months to just days.
3) Initial attempts at automated selection yielded replication parasites but optimization suppressed their emergence and enabled the selection of true nucleic acid ligands binding to targets.
In this study, kinetics of demineralization of chitin extraction from snail shells was
investigated. Chitin was extracted from snail shells by demineralizing the
deproteinized shells in 1.2 M HCl solution. Prior to demineralization, the raw snail
shells were deproteinized using 1 M NaOH solution to remove proteins and organic
matter present in the shells. The product was dried before the demineralization
process was carried out. The results showed that based on the R2 values obtained for
each of the shrinking core models considered which include; fluid film diffusion
(FFD), ash layer diffusion (ALD), and chemical reaction control (CRC), it was noted
that the CRC model was prevalent for all the various range of particle sizes analyzed
(6.3 – 4.75 mm, 4.75 – 2 mm, 2 – 1 mm, and 600 – 300 μm). The surface morphologies
and the Fourier Transform Infra-Red (FTIR) bands of the extracted chitin were
similar to previous studies
This document describes the development and validation of Neogen's Reveal Q+ lateral flow device for quantifying aflatoxin levels in grains. The device provides rapid (6 minute) and accurate results, quantifying aflatoxin from 2-150 ppb. Validation studies found the device to be highly accurate and robust across multiple operators, readers, and device lots. Additional beta site testing with industry professionals supported the device's accuracy, robustness, and ease of use for determining aflatoxin levels in corn.
The document discusses Emerald Bio's approach to parallel protein purification at the milligram scale using automated multi-target parallel processing (MTPP). Key points include:
- MTPP has delivered over 100 protein structures from over 13 targets, with over 60 containing bound ligands.
- Producing hundreds of protein structures requires thousands of purified proteins, with Emerald Bio purifying over 220 different proteins totaling over 9 grams.
- Emerald Bio's Protein Maker enables high-throughput parallel protein purification of up to 24 samples in a single run from cell lysates or fractions as small as 1 milliliter.
4. optimization of culture condition for enhanced decolorization of reactive ...Darshan Rudakiya
Many synthetic azo dyes and their metabolites are toxic, carcinogenic, and
mutagenic so removal of azo dyes using cost-effective and eco-friendly method is
major aspect.Comamonas acidovorans MTCC 3364 has been routinely reported for
different steroid bioconversion and heavy metal removal. The main purpose of this
study is to check the decolorization efficiency of Comamonas acidovorans MTCC
3364 for different dyes and to optimize the condition which gives maximum
decolorization of Reactive Orange 16 dye. The effect of various physicochemical
parameters including condition, carbon and nitrogen sources, temperature,pH and
dye concentration were studied. The % decolorization of dye was determined by
UV Visible spectroscopy. This bacterial strain efficiently decolorizes Reactive
Orange 16 at 37oC, pH 6.85 within 24 hours giving 99.03 ± 0.5 % dye
decolorization under optimum environmental conditions.
The document summarizes a study that investigated different hydrolysis methods for converting wood saw dust (WSD) into fermentable sugars for ethanol production. WSD was hydrolyzed using chemical methods with sulfuric acid (H2SO4), hydrochloric acid (HCl), and sodium hydroxide (NaOH). It was also hydrolyzed using enzymes from various fungal strains including Aspergillus fumigatus. The study found that treatment with 1N H2SO4 resulted in the highest saccharification yield of 5.52% (w/v), while enzymes from A. wentii yielded the highest saccharification of 0.119% (w/v) enzymatically.
IJERA (International journal of Engineering Research and Applications) is International online, ... peer reviewed journal. For more detail or submit your article, please visit www.ijera.com
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
Simplification of Fed-Batch Processes Using Modified Amino AcidsMerck Life Sciences
Mammalian fed-batch processes to produce biopharmaceuticals, e.g. monoclonal antibodies (mAbs), rely on strategic feeding of nutrients aiming at cell culture longevity and protein yield. At high concentrations and neutral pH, limitations in these bioprocesses arise from the low solubility or stability of some compounds, predominantly amino acids. In current processes, L-cysteine and L-tyrosine are fed separately at alkaline pH, resulting in pH peaks and precipitations. To simplify next generation processes, both amino acids have been chemically modified to enhance their respective stability and solubility profiles.
CHO fed-batch processes were substituted with the derivatives phosphotyrosine di-sodium salt (PTyr) and S-sulfocysteine sodium salt (SSC). Cellular performance as well as stability of the single substances in neutral pH feed were assessed. Lastly, the suitability of modified amino acids in fed-batch processes was confirmed examining critical quality attributes of the produced mAb.
In feed, PTyr solubility was evaluated at 70 g/L with a stability of at least 6 months stored light protected at 4 °C. The derivative was not impacting cellular performance or product quality. In cell culture supernatant, PTyr cleavage was induced by released phosphatases, thus being bioavailable for the cells.
SSC was demonstrated stable for at least 3 months in feed stored light protected at RT. In fed-batch processes, integrating the derivative into the main feed, cell specific productivity was significantly improved compared to the two-feed system. Further, IgG heterogeneity was decreased by reduced fragmentation and trisulfide bond formation of the antibody. Finally, the mechanism of action of SSC was investigated and results pointed out to an anti-oxidative response mediated through an increase in superoxide dismutase enzymes and in total intracellular glutathione pool involved in ROS elimination.
In addition to the simplification of fed-batch processes via the implementation of a single feed strategy, the two derivatives also enable the production of highly concentrated and room temperature stable feeds along with optimized space time yields.
In this webinar you will learn:
• Design of highly concentrated and stable feeds.
• Overcoming issues with unstable or insoluble amino acids.
• Understanding the function of modified amino acids in cellular metabolism and antibody production.
This document summarizes the development of efficient protocols for synthesizing 1,2,3,4-tetrahydroisoquinolin-1-ones. Several methods were developed, including the use of Mitsunobu reactions, copper-catalyzed arylations, and SNAr reactions to install various substituents on the core scaffold. These methods proved to be versatile, efficient, and amenable to parallel synthesis, allowing for SAR exploration across different regions of the molecule.
This document contains a resume for Arun Kumar, who is seeking a professionally challenging career that allows him to learn and update his skills. He has over 5 years of experience in analytical chemistry and microbiology. His experience includes chemical and microbiological analysis of water, food, and other samples using standardized testing methods. He also has experience in DNA analysis, PCR, and research related to seed and plant development. He holds an MSc in Biotechnology and a BSc in Biotechnology with first class grades.
Estimation Of The Nucleation Kinetics For The Antisolvent Crystallization Of ...cliff57
This document discusses estimating the nucleation kinetics for the anti-solvent crystallization of paracetamol in methanol-water solutions. Specifically:
- Laser back-scattering (focused beam reflectance measurement or FBRM) was used to detect nucleation events during anti-solvent crystallization experiments with paracetamol.
- Theoretical models were applied to analyze induction time and metastable zone width (MSZW) data from these experiments and estimate nucleation kinetics parameters.
- Solvent composition, in terms of the methanol-water ratio, was found to significantly impact measured induction times and MSZWs. Estimated nucleation rates also decreased with more dynamic solvent compositions.
-
This document describes a study on genetic diversity in soybean (Glycine max) using RAPD marker. Six soybean genotypes were collected and their DNA was extracted using the CTAB method. The DNA was quantified and found to range from 158-502 ng/μl. The objectives were to analyze genetic diversity among the genotypes using RAPD marker. The methodology involved DNA extraction, quantification, PCR amplification with RAPD primers, and resolving the amplified products through gel electrophoresis. The results showed good quality DNA was extracted. The study provides information on genetic variation that can aid in soybean breeding programs.
Determination Of The Crystal Growth Rate Of Paracetamol As A Function Of Solv...cliff57
1) The document discusses a study that determined the crystal growth rate of paracetamol as a function of solvent composition using antisolvent crystallization in methanol-water mixtures.
2) Crystal growth rate was found to decrease with increasing water content up to 68% water, at which point the growth rate increased.
3) The study introduced a method to link solvent composition to growth mechanism and rates. It was postulated that solubility gradient, viscosity, selective adsorption, and surface roughening affect growth rates with solvent composition.
1) The document describes a rapid method for extracting genomic DNA from filamentous fungi that involves bead beating to disrupt fungal cell walls followed by phenol-chloroform extraction and isopropanol precipitation.
2) The method yields 60-230 μg of high quality DNA per 200 mg of fungal mass within 2.5 hours without enzymatic digestion.
3) The extracted DNA was suitable for downstream PCR applications like gene amplification and RAPD analysis, demonstrating its utility for high-throughput fungal identification.
Detection of Genetically Modified Soybean in Crude Soybean Oil.PDFGordana Zdjelar
This document describes a study that aimed to detect and quantify the presence of Roundup Ready genetically modified soybean in crude soybean oil extracted from soybean seed with varying percentages of GM content. Two DNA extraction methods were compared: CTAB and a commercial DNeasy Plant Mini Kit. Real-time PCR was used to amplify DNA targets from the extracted samples to detect and quantify the GM content. The results showed that the commercial kit was superior to CTAB extraction for recovering DNA from crude oil samples. Both qualitative and quantitative PCR analysis demonstrated it is possible to monitor the GM content in crude oil derived from soybean seed with varying GM percentages.
The document describes the identification, cloning, sequencing, and characterization of the a-L-arabinofuranosidase B (abfB) gene from the phytopathogenic fungus Fusarium oxysporum f. sp. dianthi (Fod). The gene was identified using random amplified polymorphic DNA and encodes a protein of 499 amino acids. The recombinant protein expressed in E. coli had arabinofuranosidase activity and optimal activity at pH 4.0 and 50°C. Reverse transcription PCR showed that the abfB gene is actively transcribed in carnation plants infected with Fod, suggesting it plays a role in the fungus's pathogenicity.
This study demonstrates for the first time the production of recombinant human growth hormone (rhGH) from Bacillus subtilis. A hybrid gene containing the signal sequence of the B. licheniformis serine alkaline protease gene and the cDNA encoding hGH was cloned into the pMK4 plasmid and expressed under the deg promoter in B. subtilis. The rhGH produced was secreted and confirmed to have the correct mature hGH sequence. The highest rhGH concentration of 70 mg/L was obtained at 32 hours with a yield of 9 g/kg. Fermentation characteristics differed between B. subtilis containing the hybrid gene versus the plasmid alone. This expression system could be applicable for heterologous protein production from Bac
Synthesis, Molecular Docking and Antimicrobial Evaluation of New Tetrahydrobe...ijtsrd
A series of novel derivatives of Tetrahydrobenzothienopyrimidine hydrazone were synthesized and product structure was elucidated by 1NMR, C13NMR and mass spectroscopy. The synthesized compounds were evaluated against fungal and bacterial strains. The synthesized compounds showed significant antibacterial activity against Staphylococcus aureus MTCC 96, Staphylococcus pyrogenes MTCC 442, and Escherichia coli MTCC 443, Pseudomonas aeruginosa MTCC 1688 and against fungal strains Candida albicans MTCC 227, Aspergillus niger MTCC 282, Aspergillus clavatus MTCC 1323. Some derivatives showed promising result against gram positive, gram negative bacterial and fungal strains than standard drug ampicillin and grieseofulvin. In- silico molecular docking studies of the synthesized compounds was done by using GRIP batch docking method of Vlife MDS 3.0 software to study their observed activity which showed a significant correlation between the binding score and biological activity for synthesized compounds. Neetu Chopra | Kiranpreet kaur | Sanjeev Kumar "Synthesis, Molecular Docking and Antimicrobial Evaluation of New Tetrahydrobenzothienopyrimidine Derivatives" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-2 | Issue-6 , October 2018, URL: http://www.ijtsrd.com/papers/ijtsrd18756.pdf
1) The document describes an automated process for in vitro selection that was developed to generate nucleic acid aptamers faster than the traditional manual selection process.
2) An augmented Beckman Biomek 2000 pipetting robot was programmed to automate the major steps of in vitro selection, including preparing and purifying RNA, filtering RNA-target complexes, and amplifying selected sequences, in order to reduce the time needed to select aptamers from weeks or months to just days.
3) Initial attempts at automated selection yielded replication parasites but optimization suppressed their emergence and enabled the selection of true nucleic acid ligands binding to targets.
In this study, kinetics of demineralization of chitin extraction from snail shells was
investigated. Chitin was extracted from snail shells by demineralizing the
deproteinized shells in 1.2 M HCl solution. Prior to demineralization, the raw snail
shells were deproteinized using 1 M NaOH solution to remove proteins and organic
matter present in the shells. The product was dried before the demineralization
process was carried out. The results showed that based on the R2 values obtained for
each of the shrinking core models considered which include; fluid film diffusion
(FFD), ash layer diffusion (ALD), and chemical reaction control (CRC), it was noted
that the CRC model was prevalent for all the various range of particle sizes analyzed
(6.3 – 4.75 mm, 4.75 – 2 mm, 2 – 1 mm, and 600 – 300 μm). The surface morphologies
and the Fourier Transform Infra-Red (FTIR) bands of the extracted chitin were
similar to previous studies
This document describes the development and validation of Neogen's Reveal Q+ lateral flow device for quantifying aflatoxin levels in grains. The device provides rapid (6 minute) and accurate results, quantifying aflatoxin from 2-150 ppb. Validation studies found the device to be highly accurate and robust across multiple operators, readers, and device lots. Additional beta site testing with industry professionals supported the device's accuracy, robustness, and ease of use for determining aflatoxin levels in corn.
The document discusses Emerald Bio's approach to parallel protein purification at the milligram scale using automated multi-target parallel processing (MTPP). Key points include:
- MTPP has delivered over 100 protein structures from over 13 targets, with over 60 containing bound ligands.
- Producing hundreds of protein structures requires thousands of purified proteins, with Emerald Bio purifying over 220 different proteins totaling over 9 grams.
- Emerald Bio's Protein Maker enables high-throughput parallel protein purification of up to 24 samples in a single run from cell lysates or fractions as small as 1 milliliter.
4. optimization of culture condition for enhanced decolorization of reactive ...Darshan Rudakiya
Many synthetic azo dyes and their metabolites are toxic, carcinogenic, and
mutagenic so removal of azo dyes using cost-effective and eco-friendly method is
major aspect.Comamonas acidovorans MTCC 3364 has been routinely reported for
different steroid bioconversion and heavy metal removal. The main purpose of this
study is to check the decolorization efficiency of Comamonas acidovorans MTCC
3364 for different dyes and to optimize the condition which gives maximum
decolorization of Reactive Orange 16 dye. The effect of various physicochemical
parameters including condition, carbon and nitrogen sources, temperature,pH and
dye concentration were studied. The % decolorization of dye was determined by
UV Visible spectroscopy. This bacterial strain efficiently decolorizes Reactive
Orange 16 at 37oC, pH 6.85 within 24 hours giving 99.03 ± 0.5 % dye
decolorization under optimum environmental conditions.
The document summarizes a study that investigated different hydrolysis methods for converting wood saw dust (WSD) into fermentable sugars for ethanol production. WSD was hydrolyzed using chemical methods with sulfuric acid (H2SO4), hydrochloric acid (HCl), and sodium hydroxide (NaOH). It was also hydrolyzed using enzymes from various fungal strains including Aspergillus fumigatus. The study found that treatment with 1N H2SO4 resulted in the highest saccharification yield of 5.52% (w/v), while enzymes from A. wentii yielded the highest saccharification of 0.119% (w/v) enzymatically.
IJERA (International journal of Engineering Research and Applications) is International online, ... peer reviewed journal. For more detail or submit your article, please visit www.ijera.com
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
Simplification of Fed-Batch Processes Using Modified Amino AcidsMerck Life Sciences
Mammalian fed-batch processes to produce biopharmaceuticals, e.g. monoclonal antibodies (mAbs), rely on strategic feeding of nutrients aiming at cell culture longevity and protein yield. At high concentrations and neutral pH, limitations in these bioprocesses arise from the low solubility or stability of some compounds, predominantly amino acids. In current processes, L-cysteine and L-tyrosine are fed separately at alkaline pH, resulting in pH peaks and precipitations. To simplify next generation processes, both amino acids have been chemically modified to enhance their respective stability and solubility profiles.
CHO fed-batch processes were substituted with the derivatives phosphotyrosine di-sodium salt (PTyr) and S-sulfocysteine sodium salt (SSC). Cellular performance as well as stability of the single substances in neutral pH feed were assessed. Lastly, the suitability of modified amino acids in fed-batch processes was confirmed examining critical quality attributes of the produced mAb.
In feed, PTyr solubility was evaluated at 70 g/L with a stability of at least 6 months stored light protected at 4 °C. The derivative was not impacting cellular performance or product quality. In cell culture supernatant, PTyr cleavage was induced by released phosphatases, thus being bioavailable for the cells.
SSC was demonstrated stable for at least 3 months in feed stored light protected at RT. In fed-batch processes, integrating the derivative into the main feed, cell specific productivity was significantly improved compared to the two-feed system. Further, IgG heterogeneity was decreased by reduced fragmentation and trisulfide bond formation of the antibody. Finally, the mechanism of action of SSC was investigated and results pointed out to an anti-oxidative response mediated through an increase in superoxide dismutase enzymes and in total intracellular glutathione pool involved in ROS elimination.
In addition to the simplification of fed-batch processes via the implementation of a single feed strategy, the two derivatives also enable the production of highly concentrated and room temperature stable feeds along with optimized space time yields.
In this webinar you will learn:
• Design of highly concentrated and stable feeds.
• Overcoming issues with unstable or insoluble amino acids.
• Understanding the function of modified amino acids in cellular metabolism and antibody production.
This document summarizes the development of efficient protocols for synthesizing 1,2,3,4-tetrahydroisoquinolin-1-ones. Several methods were developed, including the use of Mitsunobu reactions, copper-catalyzed arylations, and SNAr reactions to install various substituents on the core scaffold. These methods proved to be versatile, efficient, and amenable to parallel synthesis, allowing for SAR exploration across different regions of the molecule.
This document contains a resume for Arun Kumar, who is seeking a professionally challenging career that allows him to learn and update his skills. He has over 5 years of experience in analytical chemistry and microbiology. His experience includes chemical and microbiological analysis of water, food, and other samples using standardized testing methods. He also has experience in DNA analysis, PCR, and research related to seed and plant development. He holds an MSc in Biotechnology and a BSc in Biotechnology with first class grades.
Proteases are enzymes that catalyze the hydrolysis of peptide bonds. They perform proteolysis, the breakdown of proteins into amino acids. There are several classes of proteases that differ in their catalytic mechanism and amino acid specificity, including serine, cysteine, aspartyl, metallo, and threonine proteases. Proteases serve many critical functions, such as protein maturation, intracellular protein degradation, regulating protein activity and localization, blood coagulation, digestion, and apoptosis. They are involved in essential biological processes and are highly regulated due to their irreversible effects on protein substrates.
1. Serine proteases use a catalytic triad of serine, histidine, and aspartate residues to hydrolyze peptide bonds through a nucleophilic attack by the serine residue.
2. Site-directed mutagenesis experiments have demonstrated the importance of these catalytic residues and the oxyanion hole for stabilizing the reaction intermediate. Mutating these residues reduces catalytic activity by several orders of magnitude.
3. Recent evidence suggests additional mechanisms such as low barrier hydrogen bonds and substrate assisted catalysis may contribute to the efficiency of serine protease catalysis.
Enzymes are proteins that catalyze chemical reactions and increase reaction rates. Several factors affect enzyme function, including enzyme concentration, substrate concentration, temperature, pH, inhibitors, activators, and salinity. Enzyme activity is highest when conditions are optimal for maintaining the enzyme's three-dimensional structure. Temperature and pH that are too high or low can cause enzymes to denature by disrupting bonds and changing their shape. Inhibitors also decrease reaction rates by competing with substrates or altering the enzyme's structure.
This document discusses various proteolytic enzymes, including their sources and applications. It describes proteases such as papain from papaya, used as a meat tenderizer, and bromelain from pineapple, also used for meat tenderizing. Pepsin from the stomach aids protein digestion. Rennin produces curdling in milk. Trypsin and cathepsins break down proteins, with trypsin used in cell culture and proteomics. These enzymes degrade proteins through hydrolysis of peptide bonds.
The document discusses factors that affect enzyme-catalyzed reactions, including substrate concentration, inhibitors, pH, temperature, and pressure. It explains how the reaction rate depends on substrate and enzyme concentrations and can be influenced by activators and inhibitors. It also describes how each enzyme has an optimal pH range and temperature, and discusses how temperature affects reaction rates, microbial growth rates, and thermal inactivation of enzymes.
The following presentation is only for quick reference. I would advise you to read the theoretical aspects of the respective topic and then use this presentation for your last minute revision. I hope it helps you..!!
Mayur D. Chauhan
Protease Enzyme Application in Food Processing Mohan Naik
This document discusses proteases, which are enzymes that break down proteins. It notes that proteases are widely distributed in biological systems and constitute over 70% of industrial enzymes. Proteases have a wide range of applications including in detergents, food processing, pharmaceuticals, and leather tanning. The document categorizes proteases based on their source (animal, plant, bacterial, fungal), proteolytic mechanism (serine, threonine, cysteine, aspartic, metallo), and pH range (acidic, neutral, alkaline). It provides examples of major industrial uses of proteases in bread making, cheese production, and soy sauce manufacturing. Protease applications also include meat tenderizing, medicine
IRJET - Production of Protease by Actinomycetes under Submerged FermentationIRJET Journal
1. Actinomycetes isolated from marine sediments were screened for protease production. Isolate B22 showed protease activity on starch casein agar plates.
2. Fermentation conditions like incubation time, temperature, pH, and inoculum age were optimized for B22 to maximize protease production.
3. Under optimized conditions of 5 days incubation at 30°C, initial pH of 5.0, and using a 6th day old inoculum, B22 produced a maximum of 46.13 U/mL of protease.
Submerged fermentation of laccase producing Streptomyces chartreusis using bo...IOSR Journals
Response surface methodology was engaged for the optimization of diverse nutritional and physical parameters for laccase production by Streptomyces chartreusis strain NBRC 12753 in the submerged fermentation process. Screening of production parameters was executed using Plackett–Burman design and the variables with statistically momentous effects on laccase production were recognized. Variables such as Cupric sulphate, Pyrogallol and Yeast extract were selected for further optimization studies using Box-Behnken design. The multiple regression coefficients (R2) had a value of 0.9606, indicating that the model could explain up to 96.06 % of the variability of the response. This methodology facilitated analysis of the experimental data to establish the optimum conditions for the process and understand the contribution of individual factors to evaluate the response under optimal conditions. Thus application of Box-Behnken approach appears to have potential usage in process application.
Optimization Of The Possibility Synthetic Nattokinase In Soybean Substrates T...inventionjournals
Nattokinase is an enzyme with strong fibrinolytic activity that can be used for preventing thrombolytic diseases. In this study, we make survey the effect fermentation conditions to B.subtilis natto strain on the soybean substrate to enhance the nattokinase activity and optimization biosynthesis capabilities of nattokinase by Plackett-Burman experimental combined with response surface methodology RSM-CCD. The optimal results received nattokinase activity 136.6 FU/g. We also try to create dried products by freeze drying process in the conditions -80°C, 24 hours, 6-7 Pa. After that we examined moisture and nattokinase activity in these product. The results of experiments show that moisture was 4.3%, nattokinase activity was 515 FU/g. This research help expand application for creating soybeans powder products with hight nattokinase activity.
Isolation, Optimization, Production and Purification of Alpha Amylase from ...IRJET Journal
This document summarizes research on the isolation, optimization, production, and purification of the enzyme alpha amylase from soil bacteria. Key points:
1) The bacteria Bacillus subtilis was isolated from soil samples and identified as an alpha amylase producer through starch hydrolysis screening and biochemical tests.
2) Fermentation conditions like pH and temperature were optimized for maximum enzyme production, with pH 7.0 and 37°C found to be optimal.
3) The enzyme was partially purified using ammonium sulfate precipitation and further purified via dialysis. SDS-PAGE was used to determine the molecular weight of the purified amylase.
The document describes optimization of the fermentation medium for production of biomass and nattokinase by Bacillus subtilis natto. Initial tests confirmed the bacterium isolated from Vietnamese natto food was Bacillus subtilis natto. Six factors in the fermentation medium were screened using Plackett-Burman design, identifying soybean peptone and CaCl2 as significant for biomass production. Response surface methodology was used to optimize the medium for highest dried cell weight of 3.033 g/L. This optimized medium increased nattokinase yield by over 30% to 31.06 FU/mL compared to the initial medium.
IJRET : International Journal of Research in Engineering and Technology is an international peer reviewed, online journal published by eSAT Publishing House for the enhancement of research in various disciplines of Engineering and Technology. The aim and scope of the journal is to provide an academic medium and an important reference for the advancement and dissemination of research results that support high-level learning, teaching and research in the fields of Engineering and Technology. We bring together Scientists, Academician, Field Engineers, Scholars and Students of related fields of Engineering and Technology
Screening, optimization of production and partial characterization of alkalin...eSAT Journals
Abstract Bacillus strains isolated from the salteren pond (Kakinada) were screened and identified for high alkaline protease activity. The isolates which were positive on skim milk agar (1%) were selected as protease producing strains. Of the ten bacterial isolates screened, isolate S-8 was observed as a potential haloalkaline protease producer and it was identified as Bacillus cereus strain S8 (MTCC NO: 11901) by 16S rRNA gene sequencing, phylogenetic tree analysis and by different biochemical tests. Protease production was enhanced by optimizing the culture conditions. The nutritional factors such as carbon and nitrogen sources, NaCl and also physical parameters like temperature, incubation time, pH, inoculum size were optimized for the maximum yield of protease. Studies on the effect of different carbon and nitrogen sources revealed that maximum protease production was obtained in the medium supplemented with Molasses,1%(w/v); Potassium nitrate, 0.75%(w/v); salt solution- 5%(v/v) {MgSo4.7H2O, 0.5%(w/v); KH2PO4, 0.5%(w/v)}; FeSO4.7H2O, 0.01%(w/v) and CaCO3, 0.5% respectively. Thus, with selected carbon and nitrogen sources along with 1 % NaCl and 2% inoculum the maximum protease production (205.0 U/ml) was obtained in the period of 72 h incubation at pH-12.0 under 160 rpm when compared to the initial enzyme production (165.0 U/ml). The crude enzyme extract of this strain was also characterized with respect to temperature, pH, incubation period and different concentrations of casein which was used as enzyme substrate. This study shows that the enzyme has wide range of pH stability from 8 to 11 with optimum activity at pH-10.0. It is thermostable with optimum activity at 70°C (392U/ml) with 1h incubation of enzyme with 1% casein as its substrate. From the above investigations it was concluded that the protease production by these microorganisms at wide temperatures and pH ranges could be explored for varied industrial applications.
Optimization of cultural conditions for lactic acid production by lactobacill...eSAT Journals
Abstract The effects on the lactic acid (LA) production by thermophile Lactobacillus bulgaricus ATTC 11842 on whey as a basal medium of seven factors namely, temperature °C, pH, Lactose g/l, Yeast extract g/l, corn steep liquor (CSL) g/l, K2HPO4 g/l, and salts g/l (MnSO4, MgSO4 and FeSO4) were investigated, through the statistical analysis of the results by Plackett and Burmann experimental design. pH was found to have the high significant effect on lactic acid production. By response surface methodology (RSM) design the optimal value of pH and concentrations in the medium of yeast extract, K2HPO4 and salts were then investigated, it should be 5.5 of pH, 2.73 g/l of K2HPO4, 1.59 g/l of yeast extract and 0.0326 g/l, 0.1304 g/l, 0.01304 g/l of MgSO4, MnSO4 and FeSO4 respectively. The results obtained with the optimal results were 20.9592 g/l. of lactic acid and the corresponding yields was 0.5665% (ratio between the amount of lactic acid produced and the initial concentration of lactose). Index Terms: Lactic acid, experimental design, Plackett and Burmann, Lactobacillus bulgaricus, whey
1. The document reports on the isolation, identification, and production of alkaline protease enzymes from Bacillus species for industrial applications.
2. Six alkaline protease producing Bacillus species were isolated from soil samples and identified using biochemical tests. The enzyme was partially purified and optimized.
3. Bacillus brevis was found to produce the highest alkaline protease levels when grown in production medium 3 at pH 9 and 37°C, with increased activity over 72 hours of incubation correlated with increased bacterial growth.
IJRET : International Journal of Research in Engineering and Technology is an international peer reviewed, online journal published by eSAT Publishing House for the enhancement of research in various disciplines of Engineering and Technology. The aim and scope of the journal is to provide an academic medium and an important reference for the advancement and dissemination of research results that support high-level learning, teaching and research in the fields of Engineering and Technology. We bring together Scientists, Academician, Field Engineers, Scholars and Students of related fields of Engineering and Technology.
The document summarizes a study that analyzed two samples of Glycine max Linn (soybean) seeds. Phytochemical analysis found various constituents including proteins, flavonoids, and phenolic compounds. Protein content was highest in the methanolic extract of sample 2. Thin layer chromatography identified several compounds in the extracts. Extracts showed antimicrobial activity against gram-positive and gram-negative bacteria, with the highest activity in the methanolic extract of sprouted sample 1.
Phytochemical analysis, protein content & antimicrobial activities of sel...eSAT Journals
Abstract Two seed samples of Glycine max Linn. (S1, S2) were purchased from two retail stores of local market. Non-sprouted and sprouted seed powder were extracted separately with methanol (100%, 50%) by cold maceration to obtain methanolic and hydroalcoholic extract of Glycine max Sample 1 was designated as MES1 and HES1 and sample 2 as MES2 and HES2 respectively. Phytochemical analysis indicated the presence of various phytoconstituents viz. phytosterols, flavonoids, phenolic compounds, tannins, carbohydrates, proteins, amino acids, fixed oils and fats etc. Thin layer chromatography study on extracts revealed the presence of a number of compounds. The protein content of these samples were studied. The protein content of samples MES1, HES1, MES2 and HES2 with respect to BSA was found to be 90.6 2μg/ml, 82μg/ml, 94.5μg/ml and 79.1μg/ml respectively. The highest among these were found to be in MES2. Sprouting enhanced the protein content of the two samples. The samples have shown antimicrobial activity at selected concentration and microbial strains (26mm) for gram negative bacteria (27mm) for gram positive bacteria. Keywords: Glycine max Linn, phytochemical constituents, TLC, antimicrobial activity, protein, methanolic extract, hydroalcoholic extract.
IRJET - Mono and Co-Digestion of Laminaria Digitata with Simulated Food W...IRJET Journal
This study examined anaerobic digestion of the seaweed Laminaria digitata both on its own (mono-digestion) and combined with simulated food waste (co-digestion) in continuous reactor experiments. Different mix ratios of L. digitata and food waste were tested in continuous stirred tank reactors over 85 days. The optimal mix ratio for highest methane production and efficiency was found to be 90% L. digitata and 10% food waste. Mono-digestion of L. digitata led to reactor failure due to accumulation of volatile fatty acids and lowered pH as organic loading rate increased. Co-digestion helped dilute inhibitory components and improved digestion stability compared to mono-digestion of L. digitata alone.
Statistical Optimization of Keratinase Production from Marine FungusIJERA Editor
To improve the yield of keratinase from marine fungus Scopulariopsis brevicaulis, different medium constituents were optimized using response surface methodology (RSM) based on central composite design (CCD). The strain produced 24.8U/mL and 36.4U/mL of keratinase activity in conventional method of optimization with glucose and soya bean meal as carbon and nitrogen sources. Response surface methodology which was applied to optimize concentrations of glucose, soya bean meal, feather powder and inoculum level, improved the productivity to 225.0U/mL. This value represents 6.18 fold increases in productivity as compared to conventional methods. Optimal parameters of the cultivation process were determined as glucose 1.52g/L, soya bean meal-1.08g/L, feather powder-1.04g/L and inoculum level-10.6%.
Effect of different parameters and storage conditions on liquid jaggery witho...eSAT Journals
Abstract Jaggery industry is a cottage industry in India. In today’s world liquid jaggery has been gaining a much importance due to its nutritional value. Studies were conducted to investigate the parameters affecting shelf life such pH, colour, brix , moisture content and range of temperatures on the concentration of reducing sugars. The variety Co86032 was selected to observe the effect of storage period on quality characteristics of liquid jaggery. Samples were stored at three different conditions i.e. room temperature(27°C), refrigeration (7°C) and high temperature (37°C) in pre-sterilized PET bottles for 90 days .In order to optimize the changes in properties physico-chemical tests were evaluated during storage. The pH decreased significantly, whereas, moisture content and reducing sugar increased significantly during storage. The changes in different attributes were significantly higher at 37°C temperature as compared to room temperature and refrigeration temperature. The results revealed that change in chemical composition was lower in case of refrigerated sample. Refrigerated sample was found more acceptable among other two samples after storage period of three months in terms of its chemical properties. Keywords: Liquid jaggery, without preservatives, Shelf life, Storage
Isolation and Characterization of Thermostable Protease Producing Bacteria fr...IOSR Journals
This study is a search for potential thermostable protease producing strain. Among nine protease
producing strains screened from soap industry effluent, one was selected as promising thermostable protease
producer and identified as Bacillus subtilis. The activity of the protease produced by this organism is stable up
to 70ºC. The optimum yield was achieved after 48 hours of culture, at 65ºC with the pH 8.0. The maximum
protease activity was observed at 65ºC and at pH 8.0.
This study assessed the enzymatic hydrolysis of proteins from the microalgae Chlorella pyrenoidosa and Spirulina sp. LEB 18 to produce protein hydrolysates. Three commercial proteases were used under different conditions to hydrolyze the microalgae proteins. The highest degrees of hydrolysis for Spirulina and Chlorella, respectively, were 55.31% and 52.9% and were obtained with 4 hours of reaction time using Protemax N200 protease. Statistical analysis showed that enzyme concentration, substrate concentration, and reaction time significantly affected the degree of hydrolysis. The results indicate it is possible to obtain protein hydrolysates with varying degrees of hydrolysis from microalgae
Microbial Production Of Alkaline Proteases And Evaluation Of Its Performances...Shafkat Shamim Rahman
A high alkaline protease producing bacterial strain was isolated and identified a local soil sample. The organism was gram positive and forms spore during adverse condition in the growth medium. After various tests it was suggested and the features agreed with the description of Bacillus subtilis. It was also identified as B. subtilis with 99.9% identity by API 50 CHB. The enzyme hydrolyses a number of proteins including azocasein which suggests that it is an extracellular alkaline protease. The experimentally determined isoelectric point was 5.1 and the optimal enzyme activity was at 60°C and at pH 8.5. The esterase preferentially hydrolyzed short-chain fatty acids. Native enzyme preparations typically showed a Michaelis constant (Km) and Vmax of 0.40mM and 12,200 U mg)-1, respectively. This microbial enzyme was partially purified by ammonium sulfate fractionation, dialysis, DEAE cellulose chromatography and electrophoretic analysis. Enzyme purity was tested by SDS-PAGE. Quantitative estimation has shown that 40mL of culture supernatant could dehair 2×1 cm of leather completely in 9 hours. In future the tanneries will use a combination of chemical and enzymatic processes. In practical applications, protease is a useful enzyme for promoting the hydrolysis of proteins and showing significant industrial applications.
Optimization of key process variables for enhanced refamycin b production in ...ijabjournal
In the present study of solid media conditions for the refamycin B yield by solid state fermentation was studied and optimized using both classical method and statistical design of experiments). Statistical analysis of the results of Plackett–Burman showed that the lower level of initial moisture , initial pH, barbital, glucose and to solid media, or increase in the concentration of xylose in the range tested, results in significant effect in refamycin B yield of AmycolatopsisrifamycinicaMTCC 14 by solid state
fermentation. The effect of change in the levels of initial moisture, initial pH, barbital, glucose and xylose
on the rfefamycin B yield was studied using central composite design methodology. Statistical analysis of
the data showed that all the independent process had significant effect on refamycin B yield. The interaction between initial moisture and initial pH, between initial moisture and barbital, between initial moisture and glucose, between initial moisture and xylose, between initial pH and xylose, between barbital and glucose, between barbital and xylose, and between glucose and xylose were significant when the response was refamycin B.
IJRET : International Journal of Research in Engineering and Technology is an international peer reviewed, online journal published by eSAT Publishing House for the enhancement of research in various disciplines of Engineering and Technology. The aim and scope of the journal is to provide an academic medium and an important reference for the advancement and dissemination of research results that support high-level learning, teaching and research in the fields of Engineering and Technology. We bring together Scientists, Academician, Field Engineers, Scholars and Students of related fields of Engineering and Technology.
Similar to Optimizing factors affecting protease production by a bacillus cereus using groundnut shell under solid substrate fermentation (20)
CHITINASE AS THE MOST IMPORTANT SECONDARY METABOLITES OF STREPTOMYCES BACTERISIJSIT Editor
This document summarizes a study on chitinase produced by Streptomyces bacteria. 310 Streptomyces isolates were obtained from soil samples collected across East Azerbaijan, Iran. DNA was extracted from the isolates and screened using PCR for a gene encoding family 19 chitinase, which shows significant antifungal activity. One isolate with strong antifungal effects was identified as S. griseous based on 16S rDNA sequencing and presence of the chitinase gene. The results suggest certain Streptomyces isolates have potential as biological control agents against fungal plant pathogens due to production of antifungal family 19 chitinase.
MOLECULAR ANALYSIS OF BACTERIAL GENE CODING CHITINASE ENZYMES, FAMILY 19 STR...IJSIT Editor
The document summarizes a study on molecular analysis of bacterial genes coding for chitinase enzymes in Streptomyces bacteria. Streptomyces are known to produce chitinase enzymes that belong to families 18 and 19 and have antifungal activity. The study isolated 310 Streptomyces bacteria from soil samples in East Azerbaijan, Iran. DNA was extracted from the isolates and PCR was performed to identify isolates containing genes for family 19 chitinases. Five isolates tested positive and one showed strong antifungal activity. The gene from this isolate was cloned and sequencing identified it as a family 19 chitinase gene with 96% similarity to Streptomyces griseus. The isolate shows potential as a
THE EFFECTS OF HELPING BACTERIA (PSEUDOMONAS SPP.) IN NITROGEN GREEN BEANS F...IJSIT Editor
Some- bacteria settle in the rhizosphere of legume plants and enhance the performance of ribosome
bacteria to nitrogen fixation and nodulation. In this paper, we used four isolated from two species of
Pseudomonas containing P.putida, P.fluorescens Chao, P.Flouresence Tabriz, P.flouresence B119 and Rhizobium
leguminosarumbv.phaseoli. In a factorial experiment with complete randomized blocks were used 5 levels of
helping bacteria(Pseudomonas spp.) and two rhizobium levels, four replicates were employed. Jamaran418
green bean was utilized as host plant. At the end, nodulation, growth and plant’s nitrogen indexes were
measured. The results showed that all above mentioned helping bacteria enhance the growth and nodulation
performance of green bean. It should be said that P.putida had the highest effect on the green bean
nodulation increase along with rhizobium (130%) followed by P.fluorescens Tabriz, P. fluorescens Chao and
P.fluorescens B119, ( 83, 63 and 17%, respectively). Also, we observed 45, 33, 22 and 8% performance
increase under the effect of P.putida, P. fluorescens Chao, P. fluorescens Tabriz and P. fluorescens B119,
respectively.
ANTIMICROBIAL PROPERTY OF AQUEOUS AND PETROLEUM ETHER LEAF EXTRACTS OF JATRO...IJSIT Editor
The experiment was carried out to investigate the antimicrobial property of aqueous and Petroleum
ether leaf extracts of Jatrophacurcas against some gram positive micro-organisms: Staphylococcus aureus,
Bacillus subtilis and some gram negative micro-organisms: Escherichia coli, Salmonella typhi using
antibiotics; Gentamycin as control. The phytochemical screening of aqueous and petroleum ether extracts
showed the presences of cardiac glycosides, steroids and terpenes, tannins, phlobatannins, anthraguinones
and saponins. The disc diffusion techniques was used to test the sensitivity of the micro-organism to the
extracts of Jatrophacurcas the results obtained show mean zones of inhibition between (19 + 0.6mm) to (30 +
0.3mm) for aqueous extract and (24 + 0.5mm) to (35 + 0.8mm) for petroleum ether extract. Micro-organisms
showed sensitivity in the following order: E.coli;(17 + 0.3mm) and (25 + 0.8mm), S.aureus; (26 + 0.2mm) and
(28 + 0.6mm), B.subtilis; (16 + 0.1mm) and (20 + 0.7mm), and S.typhi (25 + 0.2mm) and (27 + 0.6mm) for
aqueous and petroleum ether extracts respectively. The minimum inhibition concentration (MIC) for both
extracts show that the extracts inhibited the growth of the entire test organism at concentration 0.6mg/ml.
This result thus suggests the potency of Jatrophacurcas as an antimicrobial agent especially at the
concentration employed.
BIO CHEMICAL EFFECT OF 1, 5-BIS (3, 5-DIMETHYLPYRAZOL-1-YL)-3- OXAPENTANE-DIA...IJSIT Editor
The document summarizes a study that investigated the biochemical effects of 1,5-Bis(3,5-Dimethylpyrazol-1-yl)-3-oxapentane-diacetatocopper in albino rats. The study found that the compound had antidiabetic effects by lowering blood glucose levels but also caused abnormalities. Rats treated with the compound showed decreases in serum glucose and albumin levels but increases in ALT and AST levels. Long-term treatment for 6 weeks also significantly decreased body weight in treated rats. The compound affected both liver and blood biochemistry in rats.
THE EFFECT OF ALSTONEA BOONEI STEM BARK PLUS CISPLATININDUCED RENAL INSUFFIC...IJSIT Editor
The bark of Alstoniaboonei stem was analysed for the medicinal and the effect of extracts on induced
renal insufficiency. The plant material was collected in August-September 2012 and Rats 100-150g body
weights were subjected to the study. Normal saline as control, Cisplatin, and cisplatin plus Alstoneiboonei
stem bark extract were administered and the result summary for serum creatinine in cisplatin treated Rats
(2.69±0.32mg/dl) and in Rats administrates cisplastin plus Alstoniaboonei stem bark extract
(2.5±0.01mg/dl) were elevated compared to saline control (1.89±0.89mg/dl). Serum urea in cisplatin treated
Rats was (38.4 ±2.98mg/dl) compared to Rats administrates with cisplatin plus the extract (38.4±2.98mg/dl)
and saline control (24.94±3.76mg/dl). The study indicates Alstoniaboonei stem bark extract reduced the
renal insufficiency in rats.
The study was carried out to investigate the effect of the aqueous extracts of
Myristicafragrans(Nutmeg), Murrayakoenigi(curry leaf) and Aframomummelegueta(Guinea pepper) on Some
Biochemical and haematologicalParameters. Sixteen (16) wister strain rats weighing between 130 – 180g
were divided into four (4) groups of four (4) rats each and for 21 days fed the following diets: Group A –
normal diet + myristicafragrans (Nutmeg) aqueous extract, Group B – normal diet + murrayakoenigi (curry
leaf) aqueous extract, Group C – normal diet + aframomummelegueta (Guinea pepper) aqueous extract, Group
D – normal diet (control). After a period of 21 days the rats were sacrificed and the serum was taken for the
following estimations: total protein, albumin, total bilirubin, direct bilirubin, aspartate transaminase, alanine
transaminase, alkaline phosphatase, total cholesterol, triglyceride, HDL cholesterol, LDL cholesterol and
glucose. The whole blood was taken for packed cell volume and white blood cell count. The results indicated
that oral administration of myristicafragrans, murrayakoenigi and aframomummelegueta to rat’s exhibit
remarkable hypolipidaemic activity and lowering glucose concentration. The oral administration of these
three spices exhibit protein increasing activities compared with the control rats. The packed cell volume and
white cell values of all the rats decreased after feeding with experimental diet (aqueous extract) compare
with the control rats. It is clear from this study thatMyristicafragrans(Nutmeg), Murrayakoenigi(curry leaf)
andAframomummelegueta (Guinea pepper) contain significant amounts of phytochemicals and exhibit
hypolipidaemic activity when consumed.
THE INFLUENCE OF SILICONE ANTIFOAM FROM LEATHER AND DYING WASTE WATER EFFLUE...IJSIT Editor
This study investigates the influence of silicone antifoam agent on waste water from Gashash leather
and Nigerian Spinning and Dying industries (NSD). Waste water from the outlet of the industries were
collected and analyzed for physicochemical parameters. Silicone antifoam was added to the wastewater to
determine the impact of the silicone antifoam on turbidity and chemical oxygen demand (COD)
concentrations. The result shows that both turbidity and COD values significantly increased even when small
concentration of the silicone antifoam was added. Further, independent t-test was used to identify the
variance between the mean value of the wastewater from leather, spinning and dying industries, the results
indicated that there are no significant differences (observed t 0.544, critical t 2.015, and p value 0.589)
between the waste water in leather and dying industries.
WATER INTAKE CHARACTERISTICS OF DIFFERENT SOIL TYPES IN SOUTHERN BORNO NIGERIA IJSIT Editor
The water intake characteristics of soils under arable crop practice were studied with a view to
obtaining useful information for the design of irrigation and drainage system and for effective soil
management techniques. Parameters determined; infiltration, hydraulic conductivity, permeability, bulk
density, particle density, porosity and moisture content. The textural class of the soils from the three sites
was found to be clay. The result obtained indicates that infiltration was high initially but decreases later. This
may be due to the soil reaching a saturation point. On the average the infiltration rate was observed to
decrease with time. The coefficient of permeability was found to be 9.26 x 10 , 7.66 x 10 and 2.15 x 10 cm/s
for site A, B and C respectively. Information on infiltration and permeability are useful tools in irrigation and
other engineering design.
DETERMINATION OF ENGINEERING PROPERTIES OF POMEGRANATE FRUIT TO CALCULATION ...IJSIT Editor
In avoiding damage to fruit species the permissible falling height and permissible static pressure are
of great importance. The former is important in planning harvesting and handling operations, the latter in
selecting the height of transport containers. Fruits are generally transported in containers. The static and
dynamic forces which then act on the fruit will cause damage if they exceed given value. The static force may
be calculated from the weight of the fruit column being transported while the dynamic load is a consequence
of vibration caused by transport. The permitted static load for a given fruit may be determined
experimentally. In this study, physical properties of interest were determined for fresh pomegranate fruit
then calculations for the design of a suitable height were conducted based on the measured properties using
Ross and Isaacs’s theory. Maximum height for packing and storing of fresh pomegranate fruit in the box was
determined to be less than 123 cm based on a rupture force of 40.7 N.
COMPARSION OF ANTIOXIDANT POTENTIAL OF DIMOCARPUS LONGAN LOUR. EXTRACTS AND ...IJSIT Editor
The present study was carried out to evaluate antioxidant activity of Dimocarpus longan stems
extracts and also to investigate the main phytoconstituents in the bio-active extract. N-hexane,
dichloromethane, ethyl acetate and methanol 80% extract were tested for free radical scavenging activity on
model reaction with stable 2,2-diphenyl-1-picrylhydrazyl radical (DPPH). The results showed that ethyl
acetate was the most active one as antioxidant agent and phytochemical analysis of that extract revealed the
presence of triterpenes, flavonoids, tannins and carbohydrates. The results may help to discover new
chemical classes of natural antioxidant substances that could serve as selective agents for infectious diseases.
DIRECT EXPANSION GROUND SOURCE HEAT PUMPS FOR HEATING AND COOLINGIJSIT Editor
This article is an introduction to the energy problem and the possible saving that can be achieved
through improving building performance and the use of ground energy sources. The relevance and
importance of the study is discussed in the paper, which, also, highlights the objectives of the study, and the
scope of the theme. This study discusses some of the current activity in the GSHPs field. The basic system and
several variations for buildings are presented along with examples of systems in operation. Finally, the GCHP
is presented as an alternative that is able to counter much of the criticism leveled by the natural gas industry
toward conventional heat pumps. Several advantages and disadvantages are listed. Operating and installation
costs are briefly discussed.
BIOMINERALISED SILICA-NANOPARTICLES DETECTION FROM MARINE DIATOM CULTURE MEDIAIJSIT Editor
Diatoms are unicellular algae the most spectacular among the microorganisms assemble into a
micro-shell with a distinct 3-D shape and pattern of fine nanoscale features. In this investigation, we present
results; Field Emission Scanning Electron Microscopy images show the presence of ordered arrays of silica
nanoparticles. A number of diatoms with partially opened valves were observed on the surface of the diatom,
which indicates that cell contents inside of diatoms could release the nanoparticles into the culture solution.
We believe that the film forming silica nanoparticles are either released by the diatoms during reproduction
or after cell death due to bacterial action. Further research will investigate whether the silica nanoparticles
are produced intracellular and then released or whether synthesis occurs in cell culture medium. This
approach provides an environmentally friendly means for fabricating silica nanoparticles for drug delivery,
disease diagnostics, artificial opal films, decorative coatings and novel optical materials.
COMPARATIVE STUDIES ON NUTRITIONAL VALUE OF NORMAL AND TUMOR TISSUE, SARDINE...IJSIT Editor
This document summarizes a study on the nutritional value of normal and tumor tissue in Sardinella longiceps fish from India. The study found that protein levels were higher in normal tissue (29.15%) compared to tumor tissue (18.93%). Fatty acid analysis revealed higher levels of polyunsaturated fatty acids like linolenic acid in normal tissue. Vitamin A levels were also higher in normal tissue. The study analyzed minerals, amino acids, fatty acids, vitamins and other biochemical components to determine the nutritional composition and value of S. longiceps fish tissue. The results show that S. longiceps is a valuable food source for humans due to its high quality protein and balanced nutritional profile.
ANTIFUNGAL ACTIVITY OF SELECTED MEDICINAL PLANT EXTRACS AGAINST PLANT PATHOG...IJSIT Editor
The aim of this work was to find an alternative to chemical fungicides currently used in the control
plant pathogenic fungi Rhizoctoniasolani ,ColletotrichummusaeandFusariumoxysporum,. The antifungal
activity of the methanol extracts of six medicinal plants used in native medicine in Sri Lanka is reported.All
plant extracts were screened for their fungistatic, fungicidal activities and minimum inhibitory dilution (MID)
against above fungi. The media amended with methanol and recommended fungicide for respective fungal
strain were consider as negative and positive control respectively.Results showed that radial growth in all the
three tested organisms was significantly impaired (p<0.05) by the addition of the extracts in the culture
medium used. The test fungi differed in their reaction to the different extracts but on the whole, growth
inhibition increased with the concentration of each extract. The most active extracts, shows a marked effect of
the 20% methanol extracts from sweet flag with inhibition values of 91%, 86% and 84 % for F. oxysporum,R.
solani and C.muceawhereas those from wild basil inhibited the growth of the same pathogens by 89%, 84%
and 74%.The results showed minimal inhibitory concentrations (MIC) were 5 % (v/v) for sweet flag and wild
basil and 20% (v/v) for all other plant crude extracts. Out of six plants extract screened, wild basil and sweet
flag showed more than 80% fungal inhibition after 6 hour immersion and other extracts could not exceed
60% inhibition after any exposure time. The study revealed that methanol crude extract of sweet flag and
wild basil exhibit strong fungistatic and fungicidal activities against tested fungi. These results support the
potential use of these plant extracts in the management of diseases caused by tested plant pathogenic fungi.
OUTCOME OF TUNNELED CATHETERS IN HEMODIALYSIS PATIENTS: FIVE YEARS SINGLE CE...IJSIT Editor
Introduction: The tunneled hemodialysis catheters(THCs) are preferred for the patients who are expected to
poor survival and the attempts to arteriovenous fistulas (AVF) are failure. In our study,in hemodialysis
patients who are implemented tunneled catheter it is evaluated the mean duration for the catheters , their
complications and the factors which affect the period of the catheters.
Methods: At the Antalya Research and Education Center Hemodialysis Unit it is retrospectively evaluated the
data of 297 hemodialysis patients who are implemented tunneled catheter during 5 years .
Results: The mean duration time of the tunneled catheters has been 224.9+162.9 days. The duration time of
right internal jugular vein(RIJV) is considerably higher than left internal jugular vein(LIJV) and subclavian
veins (235.8+96.6 days). In diabetic hemodialysis patients, the duration time of the catheter is rather lower
than the other end stage renal disease reasons(184.4±72.1 days).
Conclusions: THCs must be considered as an alternative but not a permanent vascular access in hemodialysis
patients. Because of relatively short duration times than AVF, high infection risks and thrombosis , it must be
used only in patients who have problems with the creating permanent vascular access or patients with
expected low survival time. Moreover, it must be taken into consideration the duration time of the catheter is
low in diabetic hemodialysis patients. According to our results, catheter duration time was longer in RIJV than
in other insertion sites and RIJV must be preferred as first place to placement of THCs.
ANTIBACTERIAL ACTIVITY OF Citrus limonON Acnevulgaris (PIMPLES) IJSIT Editor
This document summarizes research on the antibacterial activity of lemon juice (Citrus limon) on Propionibacterium acnes, the bacteria that causes acne vulgaris (pimples). Samples were collected from patients and tested against lemon juice and a conventional cleanser at various concentrations. Both lemon juice and the cleanser were found to inhibit the bacteria, with lemon juice showing stronger antibacterial effects at lower concentrations. The minimum inhibitory concentration of lemon juice was found to be 60%, compared to 60% for the cleanser. At concentrations of 80% or higher, both lemon juice and the cleanser showed bactericidal effects, completely killing the bacteria. The study concludes that lemon juice is more effective than conventional clean
COMPARATIVE STUDY ON HEAVY METAL CHARACTERISTICS OF LEACHATE FROM MUNICIPAL ...IJSIT Editor
Rapid urbanization and population growth are largely responsible for very high increasing rate of
solid waste in the urban areas, its proper management and recycling is major problems of Municipal
Corporation. The analytical analysis revealed that the leachate show high concentration of heavy metals viz.,
Pb, Zn, Fe, Mn and Cu. However, their high concentration in municipal solid waste leachate may cause
contaminants for environmental pollution. Therefore, present investigation deals with analyze the heavy
metals concentration in municipal solid waste leachate.
PHARMACOGNOSTICAL AND PHYTO–CHEMICAL EVALUATION OF RAKTADUSHTIHAR YOGAIJSIT Editor
The Rakta has vital role in the maintenance of health. If Rakta is in proper quantity and having desirable
qualities too, it promotes health, improves complexion, strength and vigor. Raktadushtihara Yoga was
formulated to assess its role in the management of Raktadushti. The present study deals with the
standardization of Raktadushtihara Yoga through the Pharmacognostical and pharmaceutical standards.
Organoleptic features of coarse powder were within normal range. The pH value was 6.5, water soluble
extract 46.9% w/w, methanol soluble extract 25.9%, ash value 8.73%, loss on drying 9.63% and average
weight was 512 mg. HPTLC was carried out after organizing appropriate solvent system in which maximum 2
spots were distinguished at 254 nm.
This slide is special for master students (MIBS & MIFB) in UUM. Also useful for readers who are interested in the topic of contemporary Islamic banking.
Physiology and chemistry of skin and pigmentation, hairs, scalp, lips and nail, Cleansing cream, Lotions, Face powders, Face packs, Lipsticks, Bath products, soaps and baby product,
Preparation and standardization of the following : Tonic, Bleaches, Dentifrices and Mouth washes & Tooth Pastes, Cosmetics for Nails.
This presentation includes basic of PCOS their pathology and treatment and also Ayurveda correlation of PCOS and Ayurvedic line of treatment mentioned in classics.
Main Java[All of the Base Concepts}.docxadhitya5119
This is part 1 of my Java Learning Journey. This Contains Custom methods, classes, constructors, packages, multithreading , try- catch block, finally block and more.
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The simplified electron and muon model, Oscillating Spacetime: The Foundation...RitikBhardwaj56
Discover the Simplified Electron and Muon Model: A New Wave-Based Approach to Understanding Particles delves into a groundbreaking theory that presents electrons and muons as rotating soliton waves within oscillating spacetime. Geared towards students, researchers, and science buffs, this book breaks down complex ideas into simple explanations. It covers topics such as electron waves, temporal dynamics, and the implications of this model on particle physics. With clear illustrations and easy-to-follow explanations, readers will gain a new outlook on the universe's fundamental nature.
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Assessment and Planning in Educational technology.pptxKavitha Krishnan
In an education system, it is understood that assessment is only for the students, but on the other hand, the Assessment of teachers is also an important aspect of the education system that ensures teachers are providing high-quality instruction to students. The assessment process can be used to provide feedback and support for professional development, to inform decisions about teacher retention or promotion, or to evaluate teacher effectiveness for accountability purposes.
Optimizing factors affecting protease production by a bacillus cereus using groundnut shell under solid substrate fermentation
1. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
Optimizing factors affecting protease production by a Bacillus cereus using
groundnut shell under solid substrate fermentation
P. RATHAKRISHNAN, P. NAGARAJAN
Department of Chemical Engineering, Annamalai University, Annamalainagar-608002, Tamilnadu, India.
ABSTRACT
The physical factors affecting the production of protease from bacillus cereus was investigated. Agro
industrial waste product groundnut shells were used as substrates in solid-state fermentation for protease enzyme
production. The response surface methodology (RSM) was used to optimize protease production by implementing the
central-composite design (CCD). The physiological fermentation factors such as pH (8.0), temperature (43°C),
fermentation time (26 hrs), inoculum level (3.2 ml) and substrate concentration (9.6g) were optimized by statistical
analysis using response surface methodology. The maximum yield of protease production was 76.75U/gds. This was
evidenced by the higher value of coefficient of determination (R 2= 0.9994).
Keywords: RSM, groundnut shells, protease, CCD, bacillus cereus, physical factors optimization.
IJSIT (www.ijsit.com), Volume 1, Issue 2, November-December 2012 114
2. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
INTRODUCTION
Proteases (EC 3.4) form an important class of commercially important enzymes and find applications in
detergent, leather, food and pharmaceutical industries, constituting approximately
40% of the total enzyme market [1].
In recent years, there has been an increasing trend towards efficient utilization and value-addition of agro-
industrial residues such as coffee pulp and husk, cassava husk, cassava bagasse, sugarcane bagasse, sugar beet pulp,
apple pomace, declassified potatoes, etc. [2–7].
Bacillus strains are known to produce and secrete large quantities of extracellular enzymes and constitute a
major group of industrial enzyme producers due to the robust nature of the organism as well as its enzymes [8].
Bacterial systems are being increasingly investigated for the production of enzymes and metabolites by SSF.
Solid-state fermentation (SSF) has been defined as the fermentation process which involves solid matrix and
is carried out in absence or near absence of free water; however, the substrate must
Possess enough moisture to support growth and metabolism of the microorganism. The solid matrix could be either
the source of nutrients or simply a support impregnated by the proper nutrients that allows the development of the
microorganisms. [9]
Statistical approaches offer ideal ways for process optimization studies in biotechnology [10, 11]. Response
surface methodology (RSM) is now being routinely used for optimization studies in several biotechnological and
industrial processes [12-14].Here, we report the use of RSM to study the effects various physicochemical factors on
protease production from B. cereus
As the demand for proteases increases, it is expected that hyperactive strains will emerge and that the
enzymes produced by new strains could be used as catalysts in different industries. In the present work, the strain
employed was a strain of Bacillus sp. producing alkaline protease in SSF using agro-residue groundnut shell substrate.
In this paper, we report on factors that influence the maximization of alkaline protease production by Bacillus cereus
through SSF.
MATTERIALS AND METHODS
Microorganism and inoculums’ preparation:
Bacterial strain used in this work is well preserved in the laboratory. Bacterial strain Bacillus cereus was a
stock of the Microbial Type Culture collection Centre (MTCC), Chandigarh, India. The strain was maintained on
nutrient agar medium at 4◦C. The medium composition (g/l) was comprised off the following: Beef extract 1.0; Yeast
extract 2.0; Peptone 5.0; NaCl 5.0 and Agar 2.0. Cells were subcultered at monthly intervals.
Solid-state fermentation:
Groundnut shell was collected from local market in Panruti, Tamilnadu, India. The shells were washed
thoroughly with tap water and then dried. The dried materials obtained were milled and sieved to powder for using as
a carbon source for protease production. Fermentation was carried out in Erlenmeyer flasks (250 ml) with 10g of
IJSIT (www.ijsit.com), Volume 1, Issue 2, November-December 2012 115
3. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
Groundnut shell powder, supplemented with nutrients concentrations defined by the experimental design NaNO3
0.0386g/gds, Tryptone 0.0338g/gds, K2HPO4 0.2373g/gds and Malt extract 0.2090g/gds. Each flask was covered
with hydrophobic cotton and autoclaved at 121°C for 15 min. After cooling the flasks to room temperature, the flasks
were inoculated with 2 ml 24-h grown culture broth under sterile conditions. The contents of the flasks were well
mixed and incubated at 33±1ºC for 120 hrs.
During the preliminary screening process, the experiments are carried out for 5 days and it was found that at
the 28 hrs, the maximum production occurs. Hence experiments are carried out for 28 hrs.
Extraction of protease:
The enzyme was extracted according to the method described by Nagamine et al. (2003) [15]. Fermented
medium was mixed thoroughly with 50 mM glycine–NaOH buffer, pH 11 for 30 min and the extract was separated by
squeezing through a cloth. This process was repeated three times and extracts were pooled together and then
centrifuged. The supernatant was used as enzyme source for protease assay.
Optimization of process parameters:
A full factorial design, which includes all possible factor combinations in each of the factors, is a powerful tool
for understanding complex processes for describing factor interactions in multifactor systems. RSM is an empirical
statistical technique employed for multiple regression analysis by using quantitative data obtained from properly
designed experiments to solve multivariate equations simultaneously. The experiments with different pH, temperature,
fermentation time, innoculum size and substrate conc. were employed, simultaneously covering the spectrum of
variables for the production of protease in the central composite design. In order to describe the effects of pH,
temperature, fermentation time, inoculums size and substrate conc. on the protease production, batch experiments
were conducted. The coded values of the process parameters were determined by the following equation.
Xi X
x o (1)
i Δx
Where xi-coded value of the ith variable, Xi-uncoded value of the ith test variable and X0-uncoded value of the
ith test variable at center point.
The range and levels of individual variables are given in Table 2. The experimental design is given in Table 3,
along with experimental data and predicted responses. The regression analysis was performed to estimate the response
function as a second order polynomial.
k k 2 k -1 k
Y β 0 βi X β ii X β ij X i X j (2)
i1 i i1 i i1,i j j2
Where Y is the predicted response, βi, βj, βij are coefficients estimated from regression. They represent the
linear, quadratic and cross products of X1, X2, and X3 on response
IJSIT (www.ijsit.com), Volume 1, Issue 2, November-December 2012 116
4. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
Levels
Variable Code
-2.38 -1 0 +1 +2.38
pH A 6 7 8 9 10
Temperature ( o C) B 30 35 40 45 50
Fermentation Time (hrs) C 8 16 24 32 40
Inoculums size (ml) D 1 2 3 4 5
substrate concentration
E 3 6 9 12 15
(g)
Table 2: Levels of different process variables in coded and un-coded form for protease production independent
variable range and levels
Run A-pH B- C- D- E- Protease activity(u/gds)
. No Tempera Fermenta Inoculums substrate
Experimental Theoretical
ture tion time Size concentra
tion
1 0.00000 0.00000 0.00000 0.00000 0.00000 73.00 73.0603
2 2.37841 0.00000 0.00000 0.00000 0.00000 62.70 62.6081
3 1.00000 1.00000 -1.00000 -1.00000 1.00000 53.50 53.2997
4 -1.00000 -1.00000 1.00000 1.00000 -1.00000 50.00 49.9518
5 -2.37841 0.00000 0.00000 0.00000 0.00000 56.00 55.8666
6 0.00000 0.00000 -2.37841 0.00000 0.00000 60.00 59.8516
7 -1.00000 1.00000 -1.00000 -1.00000 1.00000 49.20 49.6277
8 1.00000 -1.00000 -1.00000 -1.00000 1.00000 62.30 62.2990
9 1.00000 1.00000 1.00000 -1.00000 1.00000 61.00 61.2235
10 1.00000 1.00000 1.00000 1.00000 1.00000 68.50 68.7547
11 1.00000 1.00000 -1.00000 -1.00000 -1.00000 53.00 53.1257
12 -1.00000 1.00000 -1.00000 1.00000 -1.00000 62.00 61.7850
IJSIT (www.ijsit.com), Volume 1, Issue 2, November-December 2012 117
6. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
35 0.00000 0.00000 0.00000 0.00000 0.00000 73.21 73.0603
36 1.00000 1.00000 1.00000 -1.00000 -1.00000 55.35 55.0307
37 1.00000 -1.00000 1.00000 -1.00000 1.00000 62.00 62.2665
38 1.00000 -1.00000 -1.00000 -1.00000 -1.00000 64.00 64.0937
39 -1.00000 1.00000 1.00000 1.00000 1.00000 66.30 66.0328
40 0.00000 -2.37841 0.00000 0.00000 0.00000 59.00 59.0473
41 -1.00000 -1.00000 -1.00000 1.00000 -1.00000 56.00 55.9656
42 -1.00000 1.00000 1.00000 -1.00000 -1.00000 59.60 59.9212
43 -1.00000 1.00000 1.00000 -1.00000 1.00000 59.70 59.6453
44 0.00000 0.00000 0.00000 -2.37841 0.00000 42.00 41.9400
45 0.00000 0.00000 0.00000 2.37841 0.00000 44.90 44.7347
46 1.00000 1.00000 -1.00000 1.00000 1.00000 62.60 62.8872
47 0.00000 2.37841 0.00000 0.00000 0.00000 65.00 64.7274
48 -1.00000 1.00000 -1.00000 -1.00000 -1.00000 56.00 55.9225
49 0.00000 0.00000 0.00000 0.00000 -2.37841 55.00 55.0838
50 1.00000 -1.00000 -1.00000 1.00000 1.00000 61.00 60.8428
Table 3: Experimental conditions and observed response values of 25 Central Composite Design
A statistical program package Design Expert 7.1.5, was used for regression analysis of the data obtained and
to estimate the coefficient of the regression equation. The equations were validated by the statistical tests called the
ANOVA analysis. The significance of each term in the Equation is to estimate the goodness of fit in each case.
Response surfaces were drawn to determine the individual and interactive effects of the test variable on the protease
production. The optimal values of the test variables were first obtained in coded units and then converted to the
uncoded units.
IJSIT (www.ijsit.com), Volume 1, Issue 2, November-December 2012 119
7. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
Protease assay:
Protease activity was determined using modified Auson–Hagihara method [16]. In this 1 ml of the enzyme
solution was added to 1 ml casein solution (1%, w/v casein solution prepared in 50 mM glycine–NaOH buffer, pH 11)
and incubated at 70ºC for 20 min. The reaction was terminated by adding 4 ml of 10% trichloroacetic acid and the
contents were filtered through a Whatman No. 1 filter paper. The filtrate absorbance was read at 280 nm using UV–
Visible spectrophotometer and the protease activity was calculated using tyrosine standard curve. One unit of alkaline
protease activity was defined as 1 µg of tyrosine liberated ml -1 under the assay conditions.
RESULT AND DICUSSION
To examine the combined effect of five different process parameters (independent variables), on the protease
production, a central composite design of 25 = 32 plus 8 centre points and (2x5 = 10) star points leading to a total of
50 experiments were performed. Equation (3) represents the mathematical model relating the protease production
and the second order polynomial coefficient for each term of the equation determined through multiple regression
analysis using the Design Expert 7.1.5.The coded values of the independent variables are given in Table 2. The
experimental and predicted values of protease production are also given in table 3.
The results were analyzed by using ANOVA i.e., analysis of variance suitable for the experimental design used
and cited in Table 4. The ANOVA of the quadratic regression model indicates the model to be significant. The Model F-
value of 2602.26 implied the model to be significant. Model F-value was calculated as a ratio of mean square
regression and mean square residual. Model P value (Prob>F) is very low [0.0500]. This reiterates that the model is
significant. The P values are used as a tool to check the significance of each of the coefficients, which in turn are
necessary to understand the pattern of the mutual interactions between the test variables. The F value and the
corresponding P values, along with the coefficient estimate are given in Table 4. The smaller the magnitude of the P,
the more significant is the corresponding coefficient. Values of P less than 0.0500 indicates the model terms to be
significant. The coefficient estimates and the corresponding P values along with the coefficient estimate are given in
table 4.The coefficients estimate and the corresponding P values suggests that, among the test variables used in the
study, A, B, C, D, E, AB, AC, AD, AE, BC, BD, BE, CD, CE, DE, A 2, B2, C2, D2, E2 are significant [where A-pH, B-
temperature, C-fermentation time, D-inoculums size and E-substrate conc.] model terms. Values greater than 0.1000
indicate the model terms are not significant.
IJSIT (www.ijsit.com), Volume 1, Issue 2, November-December 2012 120
9. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
Table 4: Analysis of Variance (ANOVA) for Response Surface Quadratic Model
Std. Dev. 0.24; R2 = 99.94%; R2(pred) 99.80%; R2(adj) 99.91%; C.V. % 0.39
Protease activity = +73.06+1.42* A+1.19 * B+0.48 * C+0.59 * D+0.13* E -1.44 * A * B-5.2 * A * C+0.29 * A * D+1.62 * A *
E+1.99 * B * C+2.76 * B * D +0.49 * B * E-0.52 * C * D+1.50 * C * E+0.65 * D * E -2.44 *A2 -1.98 * B2-2.13 * C2-
5.25 * D2-3.12 * E2
The predicted R2 of 0.9980 is in reasonable agreement with the adjusted R 2 of 0.9991. Adequate precision
measures the signal to noise ratio. A ratio greater than 4 is desirable. The fit of the model was also expressed by the
coefficient of regression R2, which was found to be 0.9994 indicating from properly that 99.94% the variability in the
response could be explained by the model. This implies that the prediction of experimental data is quite satisfactory.
The Coefficient of Variation (CV) indicates the degree of precision with which the treatments are compared. Usually,
the higher the value of the CV is, the lower the reliability of the experiment. Here a lower value of CV (0.39) indicates
greater reliability of the experiments performed. The Response surface estimation for protease production as
discussed in the previous section, the response surface methodology was used with five process variables to evaluate
their effect on the protease production. The response Eq. (3) was obtained for the protease production. To investigate
the interactive effect of two factors on the protease production, the response surface methodology was used and
three-dimensional plot was drawn. The inferences so obtained are discussed below. The interaction effects and
optimal levels of the variables were determined by plotting the response surface curves.
The 3D response surface curves are shown in Figs. 1 to 10. Figure 1 represents the interactive effect of pH
and temperature on protease production. From Fig. 1 it was inferred that with the increase in pH, the protease
production increases with the temperature. The optimum value of both the factors, viz, pH and temperature can be
analyzed by saddle point or by checking the maxima formed by the X and Y coordinates. The combined effect of pH
and fermentation time on protease production shown in Fig 2.
( U /g d s )
a c tiv ity
80
70
60
P r o te a s e
50
40
30
2.38 2.38
1.19 1.19
0.00 0.00
-1.19 -1.19
B: Temperature ( o C) A: pH
-2.38 -2.38
Figure 1: Response surface Plot for protease production from groundnut shell by Bacillus cereuss in solid state
fermentation as a function of pH and Temperature
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10. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
( U /g d s )
a c tiv ity
80
70
60
P r o te a s e
50
40
30
2.38 2.38
1.19 1.19
0.00 0.00
-1.19 -1.19
C: Fermentation Time (hrs) A: pH
-2.38 -2.38
Figure 2: Response surface Plot for protease production from groundnut shell by Bacillus cereus in solid state
( U /g d s )
fermentation as a function of pH and fermentation time
a c tiv ity
80
70
60
P r o te a s e
50
40
30
20
2.38 2.38
1.19 1.19
0.00 0.00
-1.19 -1.19
D: Inoculums size (ml) A: pH
-2.38 -2.38
Figure 3: Response surface Plot for protease production from groundnut shell by Bacillus cereus in solid state
fermentation as a function of pH and inoculums size
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11. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
( U /g d s )
a c tiv ity 80
70
60
P r o te a s e
50
40
30
20
2.38 2.38
1.19 1.19
0.00 0.00
-1.19 -1.19
E: substrate concentration (g) A: pH
-2.38 -2.38
Figure 4: Response surface Plot for protease production from groundnut shell by Bacillus cereus in solid state
fermentation as a function of pH and substrate concentration
Figure 3 depicts the interaction of pH and Inoculums size, Figure 4 shows the effect of pH and substrate
concentration on the protease production. The shape of the contour show good interaction between the pH and
substrate concentration, which is clearly illustrated in Fig. 4. The combined effect of temperature and fermentation
time was shown in the form of 3D plot in Fig. 5 Combined effect of temperature and innoculum size has been analyzed
( U /g d s )
from the CCD three-dimensional plot shown in Fig. 6.
a c tiv ity
80
70
60
P r o te a s e
50
40
30
2.38 2.38
1.19 1.19
0.00 0.00
-1.19 -1.19
C: Fermentation Time (hrs) B: Temperature ( o C)
-2.38 -2.38
Figure 5: Response surface Plot for protease production from groundnut shell by Bacillus cereus in solid state
fermentation as a function of temperature and fermentation time
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12. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
( U /g d s )
a c tiv ity
80
70
60
50
P r o te a s e
40
30
20
10
2.38 2.38
1.19 1.19
0.00 0.00
-1.19 -1.19
D: Inoculums size (ml) B: Temperature ( o C)
-2.38 -2.38
Figure 6: Response surface Plot for protease production from groundnut shell by Bacillus cereus in solid state
( U /g d s )
fermentation as a function of temperature and inoculum size
a c tiv ity
80
70
60
50
P r o te a s e
40
30
20
10
2.38 2.38
1.19 1.19
0.00 0.00
-1.19 -1.19
E: substrate concentration (g) B: Temperature ( o C)
-2.38 -2.38
Figure 7: Response surface Plot for protease production from groundnut shell by Bacillus cereus in solid state
fermentation as a function of temperature and substrate concentration
Figure 7 shows the effect of temperature and substrate concentration on the protease production. Figure 8
shows the response surface curves of protease production as a function of inoculums size and fermentation time.
Figure 9 shows the response surface curves of protease production as a function of substrate concentration and
fermentation time. Substrate concentration and fermentation time are the most important environmental parameters
influencing the protease production. Figure 10 depicts the interaction of inoculums size and substrate concentration,
where the maximum protease production of 61 U/gds was found to occur with inoculums size of 3ml and substrate
concentration of 8.5 g.
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13. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
( U /g d s )
a c tiv ity
80
70
60
P r o te a s e
50
40
30
20
2.38 2.38
1.19 1.19
0.00 0.00
-1.19 -1.19
D: Inoculums size (ml) C: Fermentation Time (hrs)
-2.38 -2.38
Figure 8: Response surface Plot for protease production from groundnut shell by Bacillus cereus in solid state
( U /g d s )
fermentation as a function of fermentation time and inoculum size
a c tiv ity
80
70
60
P r o te a s e
50
40
30
20
2.38 2.38
1.19 1.19
0.00 0.00
-1.19 -1.19
E: substrate concentration (g) C: Fermentation Time (hrs)
-2.38 -2.38
Figure 9: Response surface Plot for protease production from groundnut shell by Bacillus cereus in solid state
fermentation as a function of fermentation time and substrate concentration
IJSIT (www.ijsit.com), Volume 1, Issue 2, November-December 2012 126
14. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
( U /g d s )
a c tiv ity
80
70
60
P r o te a s e
50
40
30
20
2.38 2.38
1.19 1.19
0.00 0.00
-1.19 -1.19
E: substrate concentration (g) D: Inoculums size (ml)
-2.38 -2.38
Figure 10: Response surface Plot for protease production from groundnut shell by Bacillus cereus in solid state
fermentation as a function of inoculum size and substrate concentration
The response surfaces of mutual interactions between the variables were found to be elliptical for most cases.
The stationary point or central point is the point at which the slope of the contour is zero in all directions. The
coordinates of the central point within the highest contour levels in each of these figures will correspond to the
optimum values of the respective constituents. The optimum values drawn from these figures are in close agreement
with those obtained by optimizing the regression model Eq. (3). The sequential quadratic programming in MATLAB 7
is used to solve the second-degree polynomial regression Eq. (3). The optimum values for maximum protease
production were: pH (8.0), temperature (43°C), fermentation time (26 hrs), inoculum level (3.2 ml) and substrate
concentration (9.6g). The optimal values for the variables as predicted by MATLAB were found to be within the
design region. This shows that the model correctly explains the influence of the chosen variables on the protease
production.
Validation of the experimental model:
Validation of the experimental model was tested by carrying out the batch experiment under optimal
operation conditions pH (8.0), temperature (43°C), fermentation time (26 hrs), inoculum level (3.2 ml) and substrate
concentration (9.6g) established by the regression model. Three repeated experiments were performed and the
results are compared. The protease activity (73.00U/gds) obtained from experiments was close to the actual response
(73.06U/gds) predicted by the regression model, which proved the validity of the model.
IJSIT (www.ijsit.com), Volume 1, Issue 2, November-December 2012 127
15. P. Rathakrishnan et al., IJSIT, 2012, 1(2), 114-129
CONCLUSION
The feasibility of using an Agro-residue (groundnut shell) as possible substrate for the protease production
was studied using the response surface methodological approach. The optimum conditions for the maximum protease
production 76.75 U/gds using cassava waste are as follows: pH 8, temperature 43°C, fermentation time 26hrs,
inoculums size 3.2ml and substrate concentration 9.6g. The enzyme production in this range from this vastly available
by-product is significant.
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