Nattokinase is a potent fibrinolytic enzyme with the potential for fighting cardiovascular diseases. In this study, Bacillus subtilis natto were immobilized in the alginate – chitosan complex for fermentation of nattokinase enzyme. Six factors affecting the efficiency of immobilization cells were screened by Plackett – Burman design including: concentration of alginate, concentration of chitosan, pH of chitosan, concentration of CaCl2, added cells density, shaking time after supplementing chitosan. Results of optimization have identified two factors affecting the efficiency of cell immobilization. They are concentration of alginate (2.5%) and added cells density (approximately 5.86 million colonies per milliliter). With these two factors optimized and others kept at the normal level, immobilization efficiency reached 90.73%. After Bacillus subtilis natto had been immobilized by optimization of parameters, we conducted application for fermenting nattokinase. For 24 hours of fermentation, nattokinase enzyme activity reached 71.80 ± 0.19 FU/ml. Immobilized Bacillus subtilis natto cells were reused 6 times and on the 6 th time of reuse, nattokinase enzyme activity only decreased 2.7% in compared with the 1st reuse.
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.
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
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.
Royal jelly accelerates recovery from oral mucositisBee Healthy Farms
Great news for cancer patients who suffer from oral mucositis. However, individuals undergoing this type of intense therapy also need to consume large quantities of propolis to protect their weakened immune system.
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.
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
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.
Royal jelly accelerates recovery from oral mucositisBee Healthy Farms
Great news for cancer patients who suffer from oral mucositis. However, individuals undergoing this type of intense therapy also need to consume large quantities of propolis to protect their weakened immune system.
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.
(48)Screening and identifiation of Bacillus sp. isolated from traditional Vie...minhdaovan
Fibrinolytic enzymes produced by microorganisms have been attractive in prevention and
treatment of cardiovascular diseases (CVDs) by their low-cost and safety. This study focused
on screening for the existence of firinolytic enzymes in Vietnamese traditional fermented
soybean paste products and isolation and identifiation of related bacteria. Sixteen fermented
soybean paste samples were collected over three regions of Vietnam in which seven samples
gave the positive results on firinolytic enzyme activity. Miso (MS) and Green Chili (GC)
samples had the highest firinolytic enzyme activities (1.81 and 0.77 FU/g, respectively).
According to morphological features, four strains of bacteria were isolated and all of them
were found to produce firinolytic enzymes. The enzyme activities produced by four isolated
strains were in a range of 29.7 - 77.9 FU/g after culturing on solid state media for 24 h. The
isolated strains were identifid as Bacillus amyloliquefaciens using 16 rRNA sequence and
phylogenetic analysis with 99% similarity
ABSTRACT- Aberrant glycosylation has been recognized as hallmark of cancer. Exploiting differences in glycosylation between malignant and healthy tissues offers excellent opportunities to identify sensitive and specific cancer biomarkers. Plant lectins have demonstrated the ability to specifically agglutinate malignant transformed cells. Lectins are sugar binding proteins or glycoprotein of non-immune origin which agglutinate cells or precipitate glycol-conjugates. Some lectins shown to the anti- proliferative effect on cancer cells. A wide scope of this application of lectins is that it can be used for diagnosis as well as therapeutics of cancer. The objective of the present study was to purify a lectin from tubers of Arisaema intermedium and evaluate in vitro anti-proliferative potential towards HCT-15, a human colon cancer cell line. The present study was conceived as an offshoot to the ongoing work on lectins in our laboratory. The already reported Arisaema intermedium (AIL) lectin was purified on asialofetuin linked amino-activated silica bead matrix. The purity of the affinity purified lectin was ascertained by SDS-PAGE, pH-8.3. The lectin activity was assessed by hemagglutination and protein concentration was determined by Lowry’s method. The cytotoxicity of AIL towards HCT-15 was evaluated by MTT assay. The mechanism of anti-proliferative effect was assessed by evaluation of cell morphology, trypan blue exclusion assay, DNA fragmentation and nucleic acid content determination.
Key-words- Araceae, Arisaema, Asialofetuin, Antiproliferative effect, Apoptosis, Cytotoxicity Lectins, Mechanistic
Bioconversion of Penicillin to CephalosporinIOSR Journals
Cephalosporins are known as 3rd generation broad spectrum Beta lactam antibiotics, which can also be produced synthetically. Commonly, chemical ring expansion followed by an enzymatic removal of the phenylacetyl side chain is commonly employed to convert penicillin G into 7-aminodeacetoxycephalosporanic acid, the precursor for the manufacture of semisynthetic cephalosporins. This process requires several steps, is expensive and highly polluting. Thus there is a need to device a simple biological route to replace the chemical process. A mutant of Streptomyces clavuligerus NP1 was reported to converts Penicillin G to Deacetoxycephalosporin G (DAOG;phenylacetyl-7-aminodeacetoxycephalosporanic acid) enzymatically[5,8] . This enzyme, deacetoxycephalosporin synthase has the potential for the large scale transformation of Penicillin G to deacetoxycephalosporin. The present work studies the conditions required for efficient transformation of Penicillin G to Deacetoxycephalosporin using the wild type strain Streptomyces clavuligerus . Detection of cephalosporin was carried out using various methods. Additionally succinic acid formation was also studied as it could be used as a commercially important by product of the transformation. Deacetoxycephalosporin synthase also extracted and partially purified and characterised.
Effcet of Vernonia Amygdalina.Del Ethanolic Extract Fraction on Serum Prolact...iosrjce
IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS) is a double blind peer reviewed International Journal edited by the International Organization of Scientific Research (IOSR). The journal provides a common forum where all aspects of Agricultural and Veterinary Sciences are presented. The journal invites original papers, review articles, technical reports and short communications containing new insight into any aspect Agricultural and Veterinary Sciences that are not published or not being considered for publication elsewhere.
Enzymatic Saccharification of Lignocellulosic BiomassBiorefineryEPC™
Enzymatic Saccharification of Lignocellulosic Biomass
YOU AGREE TO INDEMNIFY BiorefineryEPCTM , AND ITS AFFILIATES, OFFICERS, AGENTS, AND EMPLOYEES AGAINST ANY CLAIM OR DEMAND, INCLUDING REASONABLE ATTORNEYS' FEES, RELATED TO YOUR USE, RELIANCE, OR ADOPTION OF THE DATA FOR ANY PURPOSE WHATSOEVER. THE DATA ARE PROVIDED BY BiorefineryEPCTM "AS IS" AND ANY EXPRESS OR IMPLIED WARRANTIES, INCLUDING BUT NOT LIMITED TO THE IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE EXPRESSLY DISCLAIMED. IN NO EVENT SHALL BiorefineryEPCTM BE LIABLE FOR ANY SPECIAL, INDIRECT OR CONSEQUENTIAL DAMAGES OR ANY DAMAGES WHATSOEVER, INCLUDING BUT NOT LIMITED TO CLAIMS ASSOCIATED WITH THE LOSS OF DATA OR PROFITS, WHICH MAY RESULT FROM ANY ACTION IN CONTRACT, NEGLIGENCE OR OTHER TORTIOUS CLAIM THAT ARISES OUT OF OR IN CONNECTION WITH THE USE OR PERFORMANCE OF THE DATA.
1. bioconversion of progesterone using aqueous two phase system by comamonas ...Darshan Rudakiya
Strains of Comamonas acidovoras have been
reported to have vital role in degradation of natural
as well as complex organic compounds.
Comamonas acidovoras MTCC 3364 has been
routinely reported for steroid bioconversion by
activated beads in aqueous system. Previously
studies observed that progesterone was converted
into AD and ADD steroids. These compounds were
higher valuable steroids which were mainly used in
different types of drugs. Novel system was used for
the bioconversion of progesterone in to AD and
ADD. Aqueous two-phase system was initially
optimized for the progesterone solubility, separation
of two phases and extraction of product steroids
from the system. Even 1% of PEG can easily
dissolved progesterone but 5% of PEG needed for
separating phases and for extraction purposes.
Beads was easily tolerate up to 20% of PEG system
even for 120 hours. Hexane used as better
extraction solvent among different type of solvent
system. Products were shown in PEG 6000 system
after 96 hours in fewer amounts. PEG 8000 system
showed products 48 hours in minor amount but
increased in 96 hours. Advantage of aqueous twophase
system in progesterone bioconversion was
better stability of progesterone, extraction of steroid
products and stability of the system.
Optimizing some conditions for spray drying in synbiotic capsule from Bacillu...inventionjournals
The study is of determination of various conditions for spray drying in producing synbiotic in the form of capsule from Bacillus subtilis natto. The experiments were conducted to examine effects of various factors such as the resistant starch-to-matodextrin ratio before drying, the inlet gas temperature and the inlet flow. The optimization experiments are administered: a ratio of wall materials to the core material is 5% (w/v) in which the ratio of resistant starch and maltodextrin is 1:9, the inlet gas temperatureis 1100C, the inlet flow is 5.60 ml/minute and the spray pressure is 2 bar. In the condition for spray drying as mentioned earlier, the initial step for trial production of synbiotic capsules was conducted with the cell density of B.subtilis natto being 8.55 ± 0.18 log(CFU/g), the activating effect of nattokinase is 518.2 FU/g and the moisture is 9.11%. During 60 days’ preservation of the product, the resulting indexes such as the cell density of B.subtilis natto in the capsule, the activating effect of nattokinase and the moisture are stable.
Validation Of Radiation Sterilization Dose For Proteases Immobilized On Aldeh...iosrphr_editor
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
Genotoxicity of Goji Berry (Lyciumbarbarum) In Vivo Mammalian Cellsinventionjournals
Lyciumbarbarum (Gojji berry) belongs to family Salonaceae which is found in China and Himalayan. This herb is used to prevent various diseases and in medical treatments as an alternative medicine being widely used for its antioxidant and revitalizing potential effects. In recent years, Gojji has become increasingly popular in Europe and North America as a "superfruit" and dietary supplement. The belief that herbal products do not bring any risk to health, is part of popular culture. However the term "natural" assigned to many products cannot assure no health risk. The aim of this study was to evaluate the possible genotoxic effects of aqueous extract of Lyciumbarbarum (Gojji berry) by micronucleus test and comet assay. Thirty Rattus norvegicus were divided into three equal groups: 1) experimental group, submitted to Gojji berry (200mg/kg orally); 2) positive control group (cyclophosphamide), and; 3) negative control group (distilled water). Micronucleus Tests were done by smear method of bone marrow cells performed after 48h for acute, and 72h for chronic exposure. The comet assay was performed on peripheral blood taken from the tail of each animal 4h, and 24h after intervention. Cytotoxicity was assessed by observing the DNA damage measuring the percentage of DNA in the tail (% DNA- measurement of the proportion of the total DNA present in the tail) and the tail moment (TM-tail length times the percentage of DNA in the tail), calculated by 100 nucleoids per animal and the presence of micronuclei in 2,000 polychromatic erythrocytes per animal. Analysis of variance (ANOVA) followed by Tukey test at 5% significance was used comparing the results. The data showed no significant difference in the frequency of DNA damage and the number of micronuclei between the experimental group and the negative control group. The results also suggest that the aqueous extract of Lyciumbarbarum (Gojji berry) at the dose of 200 mg/kg showed no genotoxic effect, which could, to a certain point, justifies its use.
Cytogenetic, Hematological and Enzymes Levels Parameters in the Biomonitoring...inventionjournals
Studies have demonstrated genotoxic effects by the presence of micronucleus in exfoliated cells from the buccal mucosa of agricultural workers exposed to pesticides. This study has assessed the genotoxic effects of pesticides on 61 agricultural workers from the state of Piauí, Brazil. 31 individuals were exposed to pesticides and 30 are from the same area, but were not involved in pesticides application. Cytogenetic damage were evaluated through micronucleus test in cells from the buccal mucosa and some parameters such as hematological and levels of enzymes. Exposed individuals exhibited cytogenetic damage with increased number of micronuclei in cells from the buccal mucosa in comparison with subjects from the control group with significant statistical difference (P < 0.01). We perceive that there is a statistically no significant (P > 0.05) increase in levels of plasmatic and eritrocytaireacetylcholinesterase and no statistically significant increase of phosphatase alkaline were detected in exposed workers in relation to the control group. No association was found in relation to smoking habits, alcohol consumption, protection utensils and the biomarkers analyzed or the biochemical analysis. Analysis of variance revealed a correlation between occupational exposure to pesticides of workers in Piauí and the presence of micronuclei (P < 0.05).
Synthesis And Evaluation of Different Numbers of Phosphonate Group Containing...inventionjournals
Oil fields were seriously affected by the mineral scale formation, mainly calcium deposition. This paper focused on the synthesis and scale prevention of oilfield scaling, especially carbonates scale and sulfate scale. Scale can cause loss of production due to blockages in the near-well formation, in the tubing or in the surface facilities. Phosphonates are widely accepted as the most effective inhibitors for calcium carbonate and calcium sulfate, also are widely used in many water treatment applications. Most good scale inhibitors should work at dosage levels between 5 and 25 ppm on most production systems depending on calcium concentration. The treatment rates can be affected by contaminants which take the scale inhibitor out of solution. Six different compounds of phosphonate containing groups have been synthesized and evaluated for scale inhibition property. Evaluation of synthetic phosphonate compounds as scale inhibitors has been done by using NACE Standard TM-0374-2001. From the results clear that one and two phosphonate group containing compounds do not show any characteristic of prevention of scales of either calcium carbonate or calcium sulphate. The compounds which have phosphonate group three or more show characteristics of prevention of both types of scales.
Genotoxicity of Goji Berry (Lyciumbarbarum) In Vivo Mammalian Cellsinventionjournals
Lyciumbarbarum (Gojji berry) belongs to family Salonaceae which is found in China and Himalayan. This herb is used to prevent various diseases and in medical treatments as an alternative medicine being widely used for its antioxidant and revitalizing potential effects. In recent years, Gojji has become increasingly popular in Europe and North America as a "superfruit" and dietary supplement. The belief that herbal products do not bring any risk to health, is part of popular culture. However the term "natural" assigned to many products cannot assure no health risk. The aim of this study was to evaluate the possible genotoxic effects of aqueous extract of Lyciumbarbarum (Gojji berry) by micronucleus test and comet assay. Thirty Rattus norvegicus were divided into three equal groups: 1) experimental group, submitted to Gojji berry (200mg/kg orally); 2) positive control group (cyclophosphamide), and; 3) negative control group (distilled water). Micronucleus Tests were done by smear method of bone marrow cells performed after 48h for acute, and 72h for chronic exposure. The comet assay was performed on peripheral blood taken from the tail of each animal 4h, and 24h after intervention. Cytotoxicity was assessed by observing the DNA damage measuring the percentage of DNA in the tail (% DNA- measurement of the proportion of the total DNA present in the tail) and the tail moment (TM-tail length times the percentage of DNA in the tail), calculated by 100 nucleoids per animal and the presence of micronuclei in 2,000 polychromatic erythrocytes per animal. Analysis of variance (ANOVA) followed by Tukey test at 5% significance was used comparing the results. The data showed no significant difference in the frequency of DNA damage and the number of micronuclei between the experimental group and the negative control group. The results also suggest that the aqueous extract of Lyciumbarbarum (Gojji berry) at the dose of 200 mg/kg showed no genotoxic effect, which could, to a certain point, justifies its use.
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.
(48)Screening and identifiation of Bacillus sp. isolated from traditional Vie...minhdaovan
Fibrinolytic enzymes produced by microorganisms have been attractive in prevention and
treatment of cardiovascular diseases (CVDs) by their low-cost and safety. This study focused
on screening for the existence of firinolytic enzymes in Vietnamese traditional fermented
soybean paste products and isolation and identifiation of related bacteria. Sixteen fermented
soybean paste samples were collected over three regions of Vietnam in which seven samples
gave the positive results on firinolytic enzyme activity. Miso (MS) and Green Chili (GC)
samples had the highest firinolytic enzyme activities (1.81 and 0.77 FU/g, respectively).
According to morphological features, four strains of bacteria were isolated and all of them
were found to produce firinolytic enzymes. The enzyme activities produced by four isolated
strains were in a range of 29.7 - 77.9 FU/g after culturing on solid state media for 24 h. The
isolated strains were identifid as Bacillus amyloliquefaciens using 16 rRNA sequence and
phylogenetic analysis with 99% similarity
ABSTRACT- Aberrant glycosylation has been recognized as hallmark of cancer. Exploiting differences in glycosylation between malignant and healthy tissues offers excellent opportunities to identify sensitive and specific cancer biomarkers. Plant lectins have demonstrated the ability to specifically agglutinate malignant transformed cells. Lectins are sugar binding proteins or glycoprotein of non-immune origin which agglutinate cells or precipitate glycol-conjugates. Some lectins shown to the anti- proliferative effect on cancer cells. A wide scope of this application of lectins is that it can be used for diagnosis as well as therapeutics of cancer. The objective of the present study was to purify a lectin from tubers of Arisaema intermedium and evaluate in vitro anti-proliferative potential towards HCT-15, a human colon cancer cell line. The present study was conceived as an offshoot to the ongoing work on lectins in our laboratory. The already reported Arisaema intermedium (AIL) lectin was purified on asialofetuin linked amino-activated silica bead matrix. The purity of the affinity purified lectin was ascertained by SDS-PAGE, pH-8.3. The lectin activity was assessed by hemagglutination and protein concentration was determined by Lowry’s method. The cytotoxicity of AIL towards HCT-15 was evaluated by MTT assay. The mechanism of anti-proliferative effect was assessed by evaluation of cell morphology, trypan blue exclusion assay, DNA fragmentation and nucleic acid content determination.
Key-words- Araceae, Arisaema, Asialofetuin, Antiproliferative effect, Apoptosis, Cytotoxicity Lectins, Mechanistic
Bioconversion of Penicillin to CephalosporinIOSR Journals
Cephalosporins are known as 3rd generation broad spectrum Beta lactam antibiotics, which can also be produced synthetically. Commonly, chemical ring expansion followed by an enzymatic removal of the phenylacetyl side chain is commonly employed to convert penicillin G into 7-aminodeacetoxycephalosporanic acid, the precursor for the manufacture of semisynthetic cephalosporins. This process requires several steps, is expensive and highly polluting. Thus there is a need to device a simple biological route to replace the chemical process. A mutant of Streptomyces clavuligerus NP1 was reported to converts Penicillin G to Deacetoxycephalosporin G (DAOG;phenylacetyl-7-aminodeacetoxycephalosporanic acid) enzymatically[5,8] . This enzyme, deacetoxycephalosporin synthase has the potential for the large scale transformation of Penicillin G to deacetoxycephalosporin. The present work studies the conditions required for efficient transformation of Penicillin G to Deacetoxycephalosporin using the wild type strain Streptomyces clavuligerus . Detection of cephalosporin was carried out using various methods. Additionally succinic acid formation was also studied as it could be used as a commercially important by product of the transformation. Deacetoxycephalosporin synthase also extracted and partially purified and characterised.
Effcet of Vernonia Amygdalina.Del Ethanolic Extract Fraction on Serum Prolact...iosrjce
IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS) is a double blind peer reviewed International Journal edited by the International Organization of Scientific Research (IOSR). The journal provides a common forum where all aspects of Agricultural and Veterinary Sciences are presented. The journal invites original papers, review articles, technical reports and short communications containing new insight into any aspect Agricultural and Veterinary Sciences that are not published or not being considered for publication elsewhere.
Enzymatic Saccharification of Lignocellulosic BiomassBiorefineryEPC™
Enzymatic Saccharification of Lignocellulosic Biomass
YOU AGREE TO INDEMNIFY BiorefineryEPCTM , AND ITS AFFILIATES, OFFICERS, AGENTS, AND EMPLOYEES AGAINST ANY CLAIM OR DEMAND, INCLUDING REASONABLE ATTORNEYS' FEES, RELATED TO YOUR USE, RELIANCE, OR ADOPTION OF THE DATA FOR ANY PURPOSE WHATSOEVER. THE DATA ARE PROVIDED BY BiorefineryEPCTM "AS IS" AND ANY EXPRESS OR IMPLIED WARRANTIES, INCLUDING BUT NOT LIMITED TO THE IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE EXPRESSLY DISCLAIMED. IN NO EVENT SHALL BiorefineryEPCTM BE LIABLE FOR ANY SPECIAL, INDIRECT OR CONSEQUENTIAL DAMAGES OR ANY DAMAGES WHATSOEVER, INCLUDING BUT NOT LIMITED TO CLAIMS ASSOCIATED WITH THE LOSS OF DATA OR PROFITS, WHICH MAY RESULT FROM ANY ACTION IN CONTRACT, NEGLIGENCE OR OTHER TORTIOUS CLAIM THAT ARISES OUT OF OR IN CONNECTION WITH THE USE OR PERFORMANCE OF THE DATA.
1. bioconversion of progesterone using aqueous two phase system by comamonas ...Darshan Rudakiya
Strains of Comamonas acidovoras have been
reported to have vital role in degradation of natural
as well as complex organic compounds.
Comamonas acidovoras MTCC 3364 has been
routinely reported for steroid bioconversion by
activated beads in aqueous system. Previously
studies observed that progesterone was converted
into AD and ADD steroids. These compounds were
higher valuable steroids which were mainly used in
different types of drugs. Novel system was used for
the bioconversion of progesterone in to AD and
ADD. Aqueous two-phase system was initially
optimized for the progesterone solubility, separation
of two phases and extraction of product steroids
from the system. Even 1% of PEG can easily
dissolved progesterone but 5% of PEG needed for
separating phases and for extraction purposes.
Beads was easily tolerate up to 20% of PEG system
even for 120 hours. Hexane used as better
extraction solvent among different type of solvent
system. Products were shown in PEG 6000 system
after 96 hours in fewer amounts. PEG 8000 system
showed products 48 hours in minor amount but
increased in 96 hours. Advantage of aqueous twophase
system in progesterone bioconversion was
better stability of progesterone, extraction of steroid
products and stability of the system.
Optimizing some conditions for spray drying in synbiotic capsule from Bacillu...inventionjournals
The study is of determination of various conditions for spray drying in producing synbiotic in the form of capsule from Bacillus subtilis natto. The experiments were conducted to examine effects of various factors such as the resistant starch-to-matodextrin ratio before drying, the inlet gas temperature and the inlet flow. The optimization experiments are administered: a ratio of wall materials to the core material is 5% (w/v) in which the ratio of resistant starch and maltodextrin is 1:9, the inlet gas temperatureis 1100C, the inlet flow is 5.60 ml/minute and the spray pressure is 2 bar. In the condition for spray drying as mentioned earlier, the initial step for trial production of synbiotic capsules was conducted with the cell density of B.subtilis natto being 8.55 ± 0.18 log(CFU/g), the activating effect of nattokinase is 518.2 FU/g and the moisture is 9.11%. During 60 days’ preservation of the product, the resulting indexes such as the cell density of B.subtilis natto in the capsule, the activating effect of nattokinase and the moisture are stable.
Validation Of Radiation Sterilization Dose For Proteases Immobilized On Aldeh...iosrphr_editor
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
Genotoxicity of Goji Berry (Lyciumbarbarum) In Vivo Mammalian Cellsinventionjournals
Lyciumbarbarum (Gojji berry) belongs to family Salonaceae which is found in China and Himalayan. This herb is used to prevent various diseases and in medical treatments as an alternative medicine being widely used for its antioxidant and revitalizing potential effects. In recent years, Gojji has become increasingly popular in Europe and North America as a "superfruit" and dietary supplement. The belief that herbal products do not bring any risk to health, is part of popular culture. However the term "natural" assigned to many products cannot assure no health risk. The aim of this study was to evaluate the possible genotoxic effects of aqueous extract of Lyciumbarbarum (Gojji berry) by micronucleus test and comet assay. Thirty Rattus norvegicus were divided into three equal groups: 1) experimental group, submitted to Gojji berry (200mg/kg orally); 2) positive control group (cyclophosphamide), and; 3) negative control group (distilled water). Micronucleus Tests were done by smear method of bone marrow cells performed after 48h for acute, and 72h for chronic exposure. The comet assay was performed on peripheral blood taken from the tail of each animal 4h, and 24h after intervention. Cytotoxicity was assessed by observing the DNA damage measuring the percentage of DNA in the tail (% DNA- measurement of the proportion of the total DNA present in the tail) and the tail moment (TM-tail length times the percentage of DNA in the tail), calculated by 100 nucleoids per animal and the presence of micronuclei in 2,000 polychromatic erythrocytes per animal. Analysis of variance (ANOVA) followed by Tukey test at 5% significance was used comparing the results. The data showed no significant difference in the frequency of DNA damage and the number of micronuclei between the experimental group and the negative control group. The results also suggest that the aqueous extract of Lyciumbarbarum (Gojji berry) at the dose of 200 mg/kg showed no genotoxic effect, which could, to a certain point, justifies its use.
Cytogenetic, Hematological and Enzymes Levels Parameters in the Biomonitoring...inventionjournals
Studies have demonstrated genotoxic effects by the presence of micronucleus in exfoliated cells from the buccal mucosa of agricultural workers exposed to pesticides. This study has assessed the genotoxic effects of pesticides on 61 agricultural workers from the state of Piauí, Brazil. 31 individuals were exposed to pesticides and 30 are from the same area, but were not involved in pesticides application. Cytogenetic damage were evaluated through micronucleus test in cells from the buccal mucosa and some parameters such as hematological and levels of enzymes. Exposed individuals exhibited cytogenetic damage with increased number of micronuclei in cells from the buccal mucosa in comparison with subjects from the control group with significant statistical difference (P < 0.01). We perceive that there is a statistically no significant (P > 0.05) increase in levels of plasmatic and eritrocytaireacetylcholinesterase and no statistically significant increase of phosphatase alkaline were detected in exposed workers in relation to the control group. No association was found in relation to smoking habits, alcohol consumption, protection utensils and the biomarkers analyzed or the biochemical analysis. Analysis of variance revealed a correlation between occupational exposure to pesticides of workers in Piauí and the presence of micronuclei (P < 0.05).
Synthesis And Evaluation of Different Numbers of Phosphonate Group Containing...inventionjournals
Oil fields were seriously affected by the mineral scale formation, mainly calcium deposition. This paper focused on the synthesis and scale prevention of oilfield scaling, especially carbonates scale and sulfate scale. Scale can cause loss of production due to blockages in the near-well formation, in the tubing or in the surface facilities. Phosphonates are widely accepted as the most effective inhibitors for calcium carbonate and calcium sulfate, also are widely used in many water treatment applications. Most good scale inhibitors should work at dosage levels between 5 and 25 ppm on most production systems depending on calcium concentration. The treatment rates can be affected by contaminants which take the scale inhibitor out of solution. Six different compounds of phosphonate containing groups have been synthesized and evaluated for scale inhibition property. Evaluation of synthetic phosphonate compounds as scale inhibitors has been done by using NACE Standard TM-0374-2001. From the results clear that one and two phosphonate group containing compounds do not show any characteristic of prevention of scales of either calcium carbonate or calcium sulphate. The compounds which have phosphonate group three or more show characteristics of prevention of both types of scales.
Genotoxicity of Goji Berry (Lyciumbarbarum) In Vivo Mammalian Cellsinventionjournals
Lyciumbarbarum (Gojji berry) belongs to family Salonaceae which is found in China and Himalayan. This herb is used to prevent various diseases and in medical treatments as an alternative medicine being widely used for its antioxidant and revitalizing potential effects. In recent years, Gojji has become increasingly popular in Europe and North America as a "superfruit" and dietary supplement. The belief that herbal products do not bring any risk to health, is part of popular culture. However the term "natural" assigned to many products cannot assure no health risk. The aim of this study was to evaluate the possible genotoxic effects of aqueous extract of Lyciumbarbarum (Gojji berry) by micronucleus test and comet assay. Thirty Rattus norvegicus were divided into three equal groups: 1) experimental group, submitted to Gojji berry (200mg/kg orally); 2) positive control group (cyclophosphamide), and; 3) negative control group (distilled water). Micronucleus Tests were done by smear method of bone marrow cells performed after 48h for acute, and 72h for chronic exposure. The comet assay was performed on peripheral blood taken from the tail of each animal 4h, and 24h after intervention. Cytotoxicity was assessed by observing the DNA damage measuring the percentage of DNA in the tail (% DNA- measurement of the proportion of the total DNA present in the tail) and the tail moment (TM-tail length times the percentage of DNA in the tail), calculated by 100 nucleoids per animal and the presence of micronuclei in 2,000 polychromatic erythrocytes per animal. Analysis of variance (ANOVA) followed by Tukey test at 5% significance was used comparing the results. The data showed no significant difference in the frequency of DNA damage and the number of micronuclei between the experimental group and the negative control group. The results also suggest that the aqueous extract of Lyciumbarbarum (Gojji berry) at the dose of 200 mg/kg showed no genotoxic effect, which could, to a certain point, justifies its use.
The Combined Effects of Omega3 Fatty Acids and NanoCurcumin Supplementation o...inventionjournals
Migraine is a chronic nerves system disease leads to considerable disabilities and affected quality of life. Neuro-inflammation play a key role in progression of migraine which mainly caused by release of inflammatory mediators including COX-2 and iNOs enzymes, TNF-α،IL-1β،IL-6 and adhesion molecule such as ICAM and VCAM.Curcumin and omega 3 fatty acids with anti-inflammatory and neuro-protective effects can reduce gene expression and production of inflammatory mediators. The combination of omega-3 and curcumin have also synergistic effects which a lower dose can induce a positive effect. The aim of present study is to determine whether omega 3 fatty acids, curcumin or combined of them as a complement treatment in migraine is helpful and the reduction gene expression and serum levels of inflammatory markers.The study will be conducted involving 80 episodic migraine patients that withstratified randomization method based on sex, gender and body mass index (BMI) are classified into 4 groups: 1) receiving omega 3 fatty acid supplement (1.8 gr/ day) and nano-curcumin supplement (80 mg/ day) 2) receiving omega 3 fatty acid supplement and curcumin placebo 3) receiving curcumin supplement and omega 3 fatty acid placebo 4) receiving omega 3 fatty acid placebo and curcumin placebo for 2 mounts. Blood samples will be collected after anthropometric parameter measuring then target biochemical parameters, gene expression and serum levels oftarget inflammatory mediators including COX-2 and iNOs enzymes, TNF-α،IL-1β،IL-6 and adhesion molecule such as ICAM and VCAM will be measured before and after the trial. The data will be statistically evaluated using the most appropriate tests.The results of current study will determine the efficacy of therapeutic effects of curcumin, omega 3 fatty acids and combined of them as a new insight to control and treatment of migraine.
Formulation and Evaluation of Fast Dissolving Tablets of Paracetamol Using Oa...inventionjournals
Paracetamol is a slightly water soluble drug belongs to BCS Class IV, used in various pain managements & in management of fever. The drug solubility was increased by solid dispersion method, in which two techniques namely physical mixing and co-grinding were tried at the ratios of 1:0.25, 1:0.5 & 1:0.75 for paracetamol to oats powder. Various parameters like pre & post compressional parameters were tested and final formula was selected based on disintegration time and in-vitro dissolution profile. Where, all the formulations were dispersed bellow 92 seconds and F6 formulation was showing 100% release at 20th minute and faster compared to the marketed formulation. F6 was prepared by co-grinding technique, at 1:0.75 paracetamol to oats powder ratio. F6 is showing zero-order drug release and mechanism of release is Super case – II transport (n = 0.9738). All the formulations were prepared using direct compression method, a conventional method of preparation
Genotoxicity Evaluation of Polystyrene Membrane with Collagen and Norbixin by...inventionjournals
The biocompatible membranes are widely applied in the medical field in order to stimulate tissue repair. The biological principle of this type of treatment is the repair and guided regeneration. In the literature, there are few reports of studies evaluating the effects and biological properties of norbixin in animal tissues. Thus, the present study was to evaluate the effect of polystyrene membrane with collagen and norbixin, through the micronucleus test and comet assay in rats, as part of the recommended test battery to evaluate the mutagenic potential. The research project was approved by CEP / FACID Protocol 069/2014. For this study, 15 rats were divided into 3 groups were used: A - the membrane was introduced into the peritoneum of the animals through a laparotomy; B - received cyclophosphamide at a dose of 50mg / kg intraperitoneally; C - were performed only one laparotomy. A peripheral blood sample was collected from the animals for conducting Comet assay and 72 hours after the start of the experiment were euthanized. It was collected bone marrow material of each rat to perform the micronucleus test. In conclusion, through the tests, the membrane is not genotoxic.
Evaluation of LdhIsozymes Following the Treatment of Methyl Parathion in the ...inventionjournals
Lactate dehydrogenase converts lactate into pyruvate and has a very important role in carbohydrate metabolism. LDH activity depends on its isozymes and their activities change under pathological conditions. The increase in LDH activity suggests the increased anaerobic conditionsunder the influence of methylparathion to meet the energy demand when the aerobic oxidation is lowered. Following the treatment of methyl parathion there was a differential percentage in increase or decrease of different isozymes in the fish Labeo rohita. There were three distinct bands during the 96h of treatment almost parallel to LDH5 band of human serum suggesting liver damage. This was also confirmed by histopathological studies.
Cytoprotective and DNA Protective Activity of Carica Papaya Leaf Extractsinventionjournals
Papaya (Carica papaya Linn) is commonly called as paw-paw and it belongs to the family Caricaceae. The properties of papaya fruit and other parts of the plant are also well known in traditional system of medicine. Papaya possess excellent medicinal properties for treatment of different ailments. These curative properties are based on the presence of phytochemical nutrients with antioxidant effect in different parts of the plant. It is considered as valuable nutraceutical fruit plant due to its biological activity and medicinal application.The present study was designed to determine the Cytoprotective and DNA protective activities of different fractions (Aqueous, Chloroform, Ethanol and Ethyl acetate extracts) of Carica papaya leaves. Cytoprotective capacity was assessed using erythrocytes, where ferrous sulphate was used to induce stress and the ability of the extracts to combat the induced stress was evaluated. The DNA protective potential against free radical-mediated oxidative stress was evaluated by a DNA damage inhibition assay involving agarose gel electrophoresis and UV spectrophotometric analysis. All the four fractions displayed significant cytoprotective effect on erythrocytes and prevented oxidative damage to DNA in presence of DNA damaging agent. Altogether, the results of our study lend pharmacological credence to the anti-cancerous and ethno medical use of this plant in traditional system of medicine and these resultscould be used to develop antimutagenic compounds for cancer therapy.
Penicillin is drug of Choice for Syphilis- Still it holds good?inventionjournals
Syphilis resistant to Tetracyclins and macrolids had well documented. Penicillin is still considered as drug of choice in the treatment of Syphilis. Treponemes resistant to Penicillin are yet to be documented. In a private STD clinic of Tirunelveli, South India, during the period of January 2010 to June 2015, 68 cases reactive for syphilis were noted. 13.2 % of false positive were identified. Remaining 59 cases, 31 lost for follow up after treatment. Among whom 12 responded well to treatment following anti-syphilitic treatment with Penicillin whereas 16 (57.14%) remained reactive for more than a year or more. The titers either persist as same or even go up. Nobody in this group are reactive for HIV. All these patients remain in latent state in this series. Nobody progressed to neuro or cardiovascular syphilis. This warrants that it is high time to look for some other better alternative to treat Syphilis or otherwise increase in the dosage may have to be considered. Even though this study comprises of small number of patients, it has to be studied in detail for a longer period at multiple centers.
Synthesis and Anti-Inflammatory activity of Sulpha/substituted 1,2-Diazolesinventionjournals
A Novel compound namely N1 (3 Nicotinoyl) 3,5 dimethyl-4-(N-4-sulfamoyl-azo)-1,2-diazoles has been synthesized by two step processes. Synthesis of N1 -4-sulfamoylphenylhydrazono-3,5-dimethyl propane-1,3-dione and sulfonamide, which interacting with 3-Nicotinoyl hydrazine to form final compound. The newly synthesized compound N1 -(3-Nicotinoyl)-3,5-dimethyl 4-(N1 -4-sulfamoyl phenyl azo) 1,2-diazoles was screened for antiinflammatory activity
Practical Guidelines to Improve Defect Prediction Model – A Reviewinventionjournals
Defect prediction models are used to pinpoint risky software modules and understand past pitfalls that lead to defective modules. The predictions and insights that are derived from defect prediction models may not be accurate and reliable if researchers do not consider the impact of experimental components (e.g., datasets, metrics, and classifiers) of defect prediction modeling. Therefore, a lack of awareness and practical guidelines from previous research can lead to invalid predictions and unreliable insights. Through case studies of systems that span both proprietary and open-source domains, find that (1) noise in defect datasets; (2) parameter settings of classification techniques; and (3) model validation techniques have a large impact on the predictions and insights of defect prediction models, suggesting that researchers should carefully select experimental components in order to produce more accurate and reliable defect prediction models.
“Desquamative Gingivitis Treated By An Antioxidant Therapy- A Case Report”inventionjournals
Desquamative gingivitis is described as an erythematous, desquamated or eroded gingival lesion. Various etiologic factors are present for the appearance of such lesions. Despite of considering etiology, treatment is oftenly provided by systemic or topical corticosteroids. Apart from steroid application, another optionable treatment is antioxidant therapy which provides rapid healing of the tissue. As antioxidants posses various advantageous properties, it can be considered as a first treatment option for desquamative gingivitis. The presented case report of desquamative gingivitis is successfully treated using systemic antioxidants in the form of commercially available „oxitard capsule‟.
Tuition Reduction Determination Using Fuzzy Tsukamotoinventionjournals
The process of determining cuts tuition for students are usually given with the same nominal. And in this paper is the determination of the pieces tuition for students who are less able to be different, depending on how much income parents and the number of children covered. For income parents who get discounted tuition fee of IDR Rp.1,500,000 and for the number of children in these families also determine the number of pieces obtained. Tsukamoto Fuzzy system is the model used in this paper. Each input variable is divided into three membership functions. In this paper, Nine Tsukamoto Fuzzy model rules have been applied. The system also provides a consequent change of parameters if the current parameter values to be changed. The smaller the parent's income, the greater the pieces obtained. The more children insured the greater the college acquired pieces.
Existance Theory for First Order Nonlinear Random Dfferential Equartioninventionjournals
In this paper, the existence of a solution of nonlinear random differential equation of first order is proved under Caratheodory condition by using suitable fixed point theorem. 2000 Mathematics Subject Classification: 34F05, 47H10, 47H4
Clinical and Laboratory Prognostic Factors in Malignant form of Mediterranean...inventionjournals
Mediterranean spotted fever (MSF) caused by Rickettsia conorii has become a significant health risk for suffering people and international travelers. In the past, overlooked as a serious disease, at present it is known that MSF was wrongly considered a benign condition. In this report, we present a set of clinical features and laboratory parameters in 55 patients (19 fatalities and 36 survivors) with malignant forms of the disease. The purpose of the study was to outline the prognostic factors of the fatal outcome in patients with malignant MSF. Based on our data, the main prediction factors for mortality in malignant MSF patients were: advanced age, delayed hospital admission, severe concomitant diseases, and failure to start or to complete appropriate antibiotic treatment. Laboratory prognostic factors in fatalities were: leukocytosis with a marked shift to the left; extremely high serum urea and creatinine levels; low levels of fibrinogen and prolongation of thrombin time. The most frequently involved organ systems of malignant cases were: central nervous system 100%, liver 92.72%, kidneys 60%, lungs 58.18%, myocardium 30.9%, and gastrointestinal tract 23.63%. The conducted histopathological investigations revealed lethal complications: encephalitis, brain edema, acute respiratory distress syndrome, non-cardiogenic lung swelling, acute myocarditis, gastrointestinal bleeding, hemorrhagicnecrotizing pancreatitis and acute renal failure.
Optimizing some conditions for spray drying in synbiotic capsule from Bacillu...inventionjournals
The study is of determination of various conditions for spray drying in producing synbiotic in the form of capsule from Bacillus subtilis natto. The experiments were conducted to examine effects of various factors such as the resistant starch-to-matodextrin ratio before drying, the inlet gas temperature and the inlet flow. The optimization experiments are administered: a ratio of wall materials to the core material is 5% (w/v) in which the ratio of resistant starch and maltodextrin is 1:9, the inlet gas temperatureis 1100C, the inlet flow is 5.60 ml/minute and the spray pressure is 2 bar. In the condition for spray drying as mentioned earlier, the initial step for trial production of synbiotic capsules was conducted with the cell density of B.subtilis natto being 8.55 ± 0.18 log(CFU/g), the activating effect of nattokinase is 518.2 FU/g and the moisture is 9.11%. During 60 days’ preservation of the product, the resulting indexes such as the cell density of B.subtilis natto in the capsule, the activating effect of nattokinase and the moisture are stable.
No, Orally Administered Hemolymph Of Limicolaria Aurora Does Not Reduce Blood...inventionjournals
The oral administration of the hemolymph of Limicolaria aurora at doses of 22.8 and 45.6 mg/kg body wt did not result in any significant difference in the systolic, diastolic, pulse pressure, mean arterial pressure and heart rate of both normotensive and adrenaline induced hypertensive wistar rats. The data from this study show the lack of antihypertensive potential associated with the oral ingestion of Limicolaria aurora hemolymph.
Correlations between Some Anthropometric Parameters, the Lipid Profile and Gl...inventionjournals
The study aimed to investigate the correlations between some anthropometric and lipid profile parameters, as well the glycated hemoglobin (HbA1c) values of Bulgarian females with type 2 Diabetes mellitus (T2DM). 212 women of Bulgarian ethnicity with T2DM of the age groups 40-60 and 61-80 years were included in the research. The anthropometric parameters: waist and hip circumferences were measured, and body mass index (BMI) and waist/hip ratio (WHR) were calculated. We also studied the lipid profile parameters: total cholesterol, triglycerides, high density lipoprotein (HDL), low density lipoprotein (LDL) and very low density lipoprotein (VLDL). A comparative study of the lipid profile criteria revealed significant differences in the levels of triglycerides, HDL-cholesterol and VLDL between the two age groups. Differences were found with regard to the values of glycated hemoglobin too. Correlations between the above anthropometric parameters and the lipid profile of patients with T2DM were examined. In patients of the age group 40-60 years a moderate strength correlation in opposite direction (negative or inverse correlation) was found between HbA1c and BMI (Pearson correlation coefficient 0.30-0.50, p<0.05). In the age group 61-80 years a low strength negative correlation was found between BMI and total cholesterol, as well as between BMI and LDL.
ASSESSMENT OF NANOPARTICULATED RESVERATROL AND LOSARTAN IN THE PROPHYLAXIS OF...inventionjournals
The study demonstrated that eight weeks after bilateral ovariectomy in female white Wistar rats, develops endothelial dysfunction of bone microvasculature and deterioration of regional blood flow in the bone, leading to the emergence of generalized osteoporosis. Nanoparticulated forms of Losartan and Nanoparticulated forms of Resveratrol, possessing endothelial protective action, effectively prevent the reduction of regional microcirculation in bone tissue, keeping it at the level of intact rats. It is allowed to maintain an adequate level of bone remodeling processes, which manifested by slowing the thinning of bone trabeculae.
ASSESSMENT OF NANOPARTICULATED RESVERATROL AND LOSARTAN IN THE PROPHYLAXIS OF...
Similar to Optimization of Bacillus Subtilis Natto Immobilization Process on Alginate – Chitosan Complex and Its Application for Nattokinase Fermentation
Biochemical and pharmacological study of biologically active preparation of p...inventionjournals
Our aim was to perform some biochemical and pharmacological studies of bioactive bovine placental preparation via digestion of cow placenta using enzyme contained in swine stomach. Amino acid compositions and contents in biologically active preparation of placenta, obtained by digestion of cow placenta with enzyme contained in swine stomach were measured by HPLC technique and it was found that contents of such amino acids as glycine, proline and lysine were highest and 9 essential amino acids, including valine, histidine, methionine, lysine, threonine, arginine, phenylalanine, leucine and isoleucine were measured. In pharmacological study, acute toxicity (LD50) of the preparation and effect of the preparation on immune response to sheep erythrocyte were investigated in white mice, weighing 18 to 20 g each. The study revealed acute toxicity (LD50) of the preparation was 60 ml per kg. Spleen index of the first and second experimental group animals treated by the preparation during both provoked and unprovoked immune responses increased by 1.2 to 3.09 times as compared to that of negative control animals, while splenocyte count elevated by 1.2 to 2.2 times than negative control animals. Higher contents of essential amino acids of the biologically active preparation of cattle placenta shows its biologically higher nutritive value, as well as pharmacological study reveals the preparation has minimal toxicity and higher effect to stimulate immune responses.
The Role of Cell Wall-Degrading Enzymes in the Development of Anthracnose Dis...Agriculture Journal IJOEAR
— The ability of Colletotrichumtruncatum CP2 in producing pectinolytic and cellulolytic enzymes was evaluated by shake flask fermentations. The results of enzymatic activity experiment indicated that PG was the first cell wall-degrading enzymes detected and the activities obtained were higher (0.24±0.10 U/mL) than other enzymes, which appeared later and in lower amount. After the cell wall was degraded by the action of PG, further degradation of the cell wall was affected by pectin methylesterases, pectin lyase, pectate lyase and cellulases. The disparity in enzymatic activity at different intervals may suggest their specific role for pathogenesis at proper timings.
Expression Purification and Immunodetection of a fusion protein Glutathione S...iosrjce
Glutathione S Transferase(GST) is an enzyme of a multi gene family which is involved in reducing
oxidative damage to cells and detoxification of Xenobiotic compounds and plays critical role in life processes.
The entire work was completely qualitative and the objective of my work was to deal with the induction,
extraction and purification of the GST fusion protein from pGEX 3X vector.In order to achieve high degree of
transformed cells,the E.Coli BL21 host strain was made competent using 0.1M CaCl2 and adding of pGEX 3X
vector into host made it transformed.With the induction of GST protein by 0.1mM IPTG,the desired protein was
purified through glutathione Cl agarose column and was detected by immunoblotting method with the use of
anti GST HRP conjugate Ab which expressed the desired protein.
Expression Purification and Immunodetection of a fusion protein Glutathione S...iosrjce
IOSR Journal of Biotechnology and Biochemistry (IOSR-JBB) covers studies of the chemical processes in living organisms, structure and function of cellular components such as proteins, carbohydrates, lipids, nucleic acids and other biomolecules, chemical properties of important biological molecules, like proteins, in particular the chemistry of enzyme-catalyzed reactions, genetic code (DNA, RNA), protein synthesis, cell membrane transport, and signal transduction. IOSR-JBB is privileged to focus on a wide range of biotechnology as well as high quality articles on genetic engineering, cell and tissue culture technologies, genetics, microbiology, molecular biology, biochemistry, embryology, cell biology, chemical engineering, bioprocess engineering, information technology, biorobotics.
In vitro experiments of prokaryotic and eukaryotic antimicrobial peptide cyto...AI Publications
These proteinaceous molecules, called antimicrobial peptides (AMPs), are a varied collection of antimicrobial peptides. The ability of AMPs to combat gut infections necessitates further study of the AMP-GI tract interaction. These peptides need to be tested in vitro for cytotoxicity before they may be considered for use in clinical infections. Using the MTT conversion assay, neutral red dye absorption assay, and a comparison to vancomycin, researchers examined the cytotoxicity of gallidermin, nisin A, natural magainin peptides, and melittin in two gastrointestinal cell types (HT29 and Caco-2). Sheep erythrocyte hemolytic activity was also studied, and the influence of AMPs on paracellular permeability was assessed using transepithelial resistance (TEER) and TEM. Gallidermin, nisin A, magainin I, magainin II, and melittin were the least cytotoxic AMPs. To our knowledge, only Melittin and NIS caused considerable hemolysis. There are two distinct ways that melittin and nisin differ in their ability to kill bacteria. It was the only AMP that had an effect on the permeability of the paracellular space. Intestinal tight junctions and cell–cell adhesion were destroyed by long-term melittin therapy, as were microvilli, cell debris, and cell–cell adhesion. Antimicrobial activity and low cytotoxicity make Gallidermin a promising therapeutic drug. The antibacterial properties of Melittin are limited, but its ability to transport poorly bioavailable medicines may be useful.
IJPCBS 2012, 2(1), 110-116 Kavya et al. ISSN: 2249-9504
110
INTERNATIONAL JOURNAL OF PHARMACEUTICAL, CHEMICAL AND BIOLOGICAL SCIENCES
Available online at www.ijpcbs.com
ISOLATION AND SCREENING OF STREPTOMYCES SP. FROM
CORINGA MANGROVE SOILS FOR ENZYME PRODUCTION AND
ANTIMICROBIAL ACTIVITY
M. Kavya Deepthi1*, M. Solomon Sudhakar1 and M. Nagalakshmi Devamma2 1Department of Biotechnology, Rajalakshmi Engineering College, Thandalam, 2Department of Botany, Sri Venkateswara University, Tirupati, Andhra Pr Taadmesihln, aInddui,a I.n dia.
Standardized cells with completely predictable and controllable behavior is a key focus of synthetic biology. Recent advances such as the implantation of a synthesized genome into a DNA-free bacterial shell at the J. Craig Venter Institute marked the dawn of these whole-genome scale projects. A first step towards predictable synthetic minimal cells is made through further reduction of the cellular complexity by minimizing the bacterial genome. This has lead to a short list of essential genes for cells to still be considered living entities. Although these minimal cells increase our fundamental understanding of key cellular processes, there are other important criteria to consider for synthetic cells to become biotechnologically relevant.
In this colloquium I will address the current state of research concerning biotechnological useful cells that can be used as a platform for operating precisely designed and independent biological systems. Top-down methods for the design of these organisms will be reviewed, highlighting remarkable experiments.
The purpose of the research was to explore different ways through which an existing chitin digestion protocol could be improved
Chitin is a naturally abundant mucopolysaccharide commonly found in crustacean shells, insect shells, and fungal cell walls. Chitin is known to be highly insoluble in both water and organic solvents which presents a challenge both in measuring small masses of chitin and in digestion procedures.
Application of proteomics for identification of abiotic stress tolerance in c...Vivek Zinzala
It is the study of “Proteome”.
The word "proteome" is a blend of "protein" and "genome”.
Large scale study of Proteins.
Particularly their structures and functions.
Study of full set of proteins in a cell type or tissue, and changes during various conditions
Isolation and screening of cellulolytic bacteria inhabiting different environments and optimization of cellulase production
Similar to Optimization of Bacillus Subtilis Natto Immobilization Process on Alginate – Chitosan Complex and Its Application for Nattokinase Fermentation (20)
Understanding Inductive Bias in Machine LearningSUTEJAS
This presentation explores the concept of inductive bias in machine learning. It explains how algorithms come with built-in assumptions and preferences that guide the learning process. You'll learn about the different types of inductive bias and how they can impact the performance and generalizability of machine learning models.
The presentation also covers the positive and negative aspects of inductive bias, along with strategies for mitigating potential drawbacks. We'll explore examples of how bias manifests in algorithms like neural networks and decision trees.
By understanding inductive bias, you can gain valuable insights into how machine learning models work and make informed decisions when building and deploying them.
Water billing management system project report.pdfKamal Acharya
Our project entitled “Water Billing Management System” aims is to generate Water bill with all the charges and penalty. Manual system that is employed is extremely laborious and quite inadequate. It only makes the process more difficult and hard.
The aim of our project is to develop a system that is meant to partially computerize the work performed in the Water Board like generating monthly Water bill, record of consuming unit of water, store record of the customer and previous unpaid record.
We used HTML/PHP as front end and MYSQL as back end for developing our project. HTML is primarily a visual design environment. We can create a android application by designing the form and that make up the user interface. Adding android application code to the form and the objects such as buttons and text boxes on them and adding any required support code in additional modular.
MySQL is free open source database that facilitates the effective management of the databases by connecting them to the software. It is a stable ,reliable and the powerful solution with the advanced features and advantages which are as follows: Data Security.MySQL is free open source database that facilitates the effective management of the databases by connecting them to the software.
NO1 Uk best vashikaran specialist in delhi vashikaran baba near me online vas...Amil Baba Dawood bangali
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Hierarchical Digital Twin of a Naval Power SystemKerry Sado
A hierarchical digital twin of a Naval DC power system has been developed and experimentally verified. Similar to other state-of-the-art digital twins, this technology creates a digital replica of the physical system executed in real-time or faster, which can modify hardware controls. However, its advantage stems from distributing computational efforts by utilizing a hierarchical structure composed of lower-level digital twin blocks and a higher-level system digital twin. Each digital twin block is associated with a physical subsystem of the hardware and communicates with a singular system digital twin, which creates a system-level response. By extracting information from each level of the hierarchy, power system controls of the hardware were reconfigured autonomously. This hierarchical digital twin development offers several advantages over other digital twins, particularly in the field of naval power systems. The hierarchical structure allows for greater computational efficiency and scalability while the ability to autonomously reconfigure hardware controls offers increased flexibility and responsiveness. The hierarchical decomposition and models utilized were well aligned with the physical twin, as indicated by the maximum deviations between the developed digital twin hierarchy and the hardware.
We have compiled the most important slides from each speaker's presentation. This year’s compilation, available for free, captures the key insights and contributions shared during the DfMAy 2024 conference.
Using recycled concrete aggregates (RCA) for pavements is crucial to achieving sustainability. Implementing RCA for new pavement can minimize carbon footprint, conserve natural resources, reduce harmful emissions, and lower life cycle costs. Compared to natural aggregate (NA), RCA pavement has fewer comprehensive studies and sustainability assessments.
Hybrid optimization of pumped hydro system and solar- Engr. Abdul-Azeez.pdffxintegritypublishin
Advancements in technology unveil a myriad of electrical and electronic breakthroughs geared towards efficiently harnessing limited resources to meet human energy demands. The optimization of hybrid solar PV panels and pumped hydro energy supply systems plays a pivotal role in utilizing natural resources effectively. This initiative not only benefits humanity but also fosters environmental sustainability. The study investigated the design optimization of these hybrid systems, focusing on understanding solar radiation patterns, identifying geographical influences on solar radiation, formulating a mathematical model for system optimization, and determining the optimal configuration of PV panels and pumped hydro storage. Through a comparative analysis approach and eight weeks of data collection, the study addressed key research questions related to solar radiation patterns and optimal system design. The findings highlighted regions with heightened solar radiation levels, showcasing substantial potential for power generation and emphasizing the system's efficiency. Optimizing system design significantly boosted power generation, promoted renewable energy utilization, and enhanced energy storage capacity. The study underscored the benefits of optimizing hybrid solar PV panels and pumped hydro energy supply systems for sustainable energy usage. Optimizing the design of solar PV panels and pumped hydro energy supply systems as examined across diverse climatic conditions in a developing country, not only enhances power generation but also improves the integration of renewable energy sources and boosts energy storage capacities, particularly beneficial for less economically prosperous regions. Additionally, the study provides valuable insights for advancing energy research in economically viable areas. Recommendations included conducting site-specific assessments, utilizing advanced modeling tools, implementing regular maintenance protocols, and enhancing communication among system components.
Sachpazis:Terzaghi Bearing Capacity Estimation in simple terms with Calculati...Dr.Costas Sachpazis
Terzaghi's soil bearing capacity theory, developed by Karl Terzaghi, is a fundamental principle in geotechnical engineering used to determine the bearing capacity of shallow foundations. This theory provides a method to calculate the ultimate bearing capacity of soil, which is the maximum load per unit area that the soil can support without undergoing shear failure. The Calculation HTML Code included.
Sachpazis:Terzaghi Bearing Capacity Estimation in simple terms with Calculati...
Optimization of Bacillus Subtilis Natto Immobilization Process on Alginate – Chitosan Complex and Its Application for Nattokinase Fermentation
1. International Journal of Pharmaceutical Science Invention
ISSN (Online): 2319 – 6718, ISSN (Print): 2319 – 670X
www.ijpsi.org Volume 5 Issue 3 ‖ June 2016 ‖ PP.25-30
www.ijpsi.org 25 | P a g e
Optimization of Bacillus Subtilis Natto Immobilization Process on
Alginate – Chitosan Complex and Its Application for Nattokinase
Fermentation
Hien T. Ho1
, Huong T. Nguyen2
1,2
(Department Of Biotechnology – Ho Chi Minh City University Of Technology)
ABSTRACT: Nattokinase is a potent fibrinolytic enzyme with the potential for fighting cardiovascular diseases.
In this study, Bacillus subtilis natto were immobilized in the alginate – chitosan complex for fermentation of
nattokinase enzyme. Six factors affecting the efficiency of immobilization cells were screened by Plackett –
Burman design including: concentration of alginate, concentration of chitosan, pH of chitosan, concentration of
CaCl2, added cells density, shaking time after supplementing chitosan. Results of optimization have identified
two factors affecting the efficiency of cell immobilization. They are concentration of alginate (2.5%) and added
cells density (approximately 5.86 million colonies per milliliter). With these two factors optimized and others
kept at the normal level, immobilization efficiency reached 90.73%. After Bacillus subtilis natto had been
immobilized by optimization of parameters, we conducted application for fermenting nattokinase. For 24 hours
of fermentation, nattokinase enzyme activity reached 71.80 ± 0.19 FU/ml. Immobilized Bacillus subtilis natto
cells were reused 6 times and on the 6th
time of reuse, nattokinase enzyme activity only decreased 2.7% in
compared with the 1st
reuse.
Keywords: Bacillus subtilis natto, immobilized cells, alginate, chitosan, nattokinase
I. Introduction
Nattokinase [EC 3.4.21] is an extracellular serine protease, this enzyme was demonstrated being a fibrinolytic
enzyme and considered a potential agent in the effective treatment of obstructive vascular disease [1]. This
enzyme has been discovered from various food sources such as Natto of Japanese [1], Doen-jang of Korea or
Dauchi of Chinese [2] and from various microbial especially the strains Bacillus subtilis bacteria in traditional
fermented foods from soybean [3]. Bacillus subtilis natto strains are capable of the highest yield of nattokinase
biosynthesis.
The technique to immobilize cells and enzyme by entrapment in matrix method has been applied for high
efficiency in the food industrial, pharmaceutical and environmental treatment [4]. Among the carriers applied in
the immobilization process, alginate and chitosan are widely used because these carriers are natural polymers
and have some advantages: Entrapment within gels of natural polymer is very mild technique and damage to
living cells can be minimized, easily handling, inexpensive and safe for users [5]. However, the immobilization
of cells in alginate gel has the disadvantage that immobilized bacteria are surrounded by a gel network which
strongly limits their movement. When growth occurs, bacteria push the gel network away and colonies
containing densely packed bacteria are formed. As the colony expands, it may eventually reach the surface of
the gel bead. This may lead to an eruption of the colony to the surrounding medium and limiting the reuse.
Moreover, alginate gel beads are unstable in the medium containing citrate and phosphate that are often used as
a buffer solution in the fermentation broth. Besides, the immobilized of cells in chitosan has some disadvantage
due to the small gel pore size which limits the diffusion of nutrients and metabolic products between the
fermentation broth and cells in the gel bead. In order to overcome this limitation, bacteria will be entrapped in
calcium alginate gel beads, after that they are coated with chitosan. The combination of the amino group of
chitosan and carbonate group of alginate will make up the gel network strength, which will help reduce cells
released and increase the number of cell reuse [6,7]. In addition to that, chitosan’s characteristics of antibacterial
in the fermentation broth and stability at high temperatures will create favorable conditions for Bacillus subtilis
natto cells to grow strongly [8].
II. Material And Method
1.1 Microorganism Strain And Cultural Medium
Bacillus subtilis natto was selected from bacteria collection of Ho Chi Minh University of Technology.
Bacillus subtilis natto was cultured in the nutrition broth (NB) and nutrition agar (NA), the pH 7.5, the
temperature at 370
C, the agitation of 200 rpm.
Medium for fermentation broth composed of Glucose 5.6 (g/l), Peptone 13 (g/l), MgSO4.7H2O 0.875 (g/l), NaCl
2.5 (g/l), CaCl2 0.025 (g/l) [9].
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Six factors effecting on efficiency of immobilization: concentration of alginate (1.5-3.5%), concentration of
chitosan (0.2-0.6%), pH of chitosan (5-6); concentration of CaCl2 (1-3%), initial supplement cells density (10 -
100 million colony forming units per milliliter); shaking time when supplementing chitosan (20-30 minutes)
1.2 Methods
2.2.1 Optimization process of Bacillus subtilis natto
- Screening the factors effecting on the efficiency immobilization of cell.
Six variables were examined in the Plackett Burman matrix with different 12 runs. We determined the
performance of immobilization for each validation formula and analyzed the factors having great impacts on
efficiency of immobilization cells. The main effecting factors had p value < 0.05. The chosen factors were
designed to determine the relationship between those and the response.
- Optimizing parameter for cells immobilization process
With the selected factors from the screening experiments, we carried out the initial experiment with original
values (-1,+1). Based on “Lack of fit” test, we determine the relationship between the chosen factors and
response. After analyzing the initial experiments, we determined whether the factors having great impacts on the
high regression equation are suitable or not. If the regression is linear function, we design the steepest
experiments to identify the suitable range of factors.
Based on that, we conducted the experiments for response surface methodology having the central composite
designs and determined function of the poly - nominal regression accurately to describe relationship between the
efficiency of immobilization and each factors.
The optimized results of response surface methodology were calculated by software simulation to determine the
highest immobilization efficiency and immobilization conditions.
Analyzing data and identifying the regression were carried out by Minitab 16.
2.2.2 Using immobilization of cells for fermentation nattokinase
We performed immobilization of cells with optimal parameters found from optimizing experiment then
conducted fermenting enzyme nattokinase.
Immobilized cells were fermentation with: broth being medium for fermentation, temperature at 370
C, agitation
at 200 rpm. The survey conducted on process of enzyme nattokinase biosynthesis was based on time of
fermentation by immobilization of cells. Survey process started at 8th
hour of fermentation and on every 4 hours,
sample was taken to determine enzyme nattokinase activity. Fermentation was stopped at 28th
hour.
Survey reused immobilized cells by stopping fermentation at 24th
hour, collecting broth for determine
nattokinase activity and transferring immobilized cells to fresh broth medium.
2.2.3 Analyzing nattokinase activity
Nattokinase activity was determined by ability to hydrolyze fibrin fiber. Bacillus subtilis natto fermentation was
stopped after 20 hours and broth was centrifuged at 13000 rpm for 20 minutes and obtained the supernatant to
determine nattokinase activity.
Tris-HCl (50mM, pH 7.5) of 1.3 mL and 0.4 mL 0f 0.72% (w/v) fibrinogen solution were taken in vials and kept
in water bath (370
C) for 5 minutes. Then 0.1 mL thrombin (20U/mL) was added and kept in water bath (370
C)
for 10 minutes. To this clot, 0.1 mL of enzyme was added. After incubation (370
C, 60 minutes), 2 mL of 0.2 M
trichloroacetic acid (TCA) was added. Vials were kept 20 minutes and centrifuged at 3000 x g for 5 minutes.
One unit enzyme activity is defined as the amount of enzyme required to produce an increase in absorbance
equal to 0.01 in 60 minutes at 280 nm [10].
III. Result And Discussion
1.3 Screening main effective factors of the cell immobilization efficiency
The cell immobilization efficiency and the effective values obtained from Plackett Burman matrix by
experiments were illustrated in Table1. Two factors notably influenced in cell immobilization efficiency
(p<0.05) were alginate concentration and cell density.
The alginate concentration factor and the added cells density had effect value (39.9, p = 0.01) and (45.56, 0.01)
respectively. The remaining factors had not effect significantly in designed experiments.
Table 1: The factor in Plackett Burman matrix and its effect to immobilization cells
Name of factors Symbols
of factor
Values of factors Main
effect
P values
Low (-1) High (+1)
Alginate concentration (%) X1 1.5 3.5 39.9 0.01
Chitosan concentration (%) X2 0.2 0.6 0.1 0.767
pH chitosan X3 5 6 0.09 0.774
CaCl2 concentration (%) X4 1 3 5.61 0.064
The added cells density (million X5 10 100 45.56 0.01
3. Optimization Of Bacillus Subtilis Natto Immobilization Process On Alginate – Chitosan Complex..
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colonies/ml)
Shaking time X6 20 40 5.75 0.062
R-sq = 95.10%
1.4 The optimal values of parameters in immobilizing cell process for highest efficiency of immobilization
We determined the relationship between cell immobilization efficiency and two main effective factors through 9
initial experiments. Through initial experiments, we found that alginate concentration has greatest impact on
immobilization efficiency than added cell. Through the levels of two variable and their effect on immobilization
efficiency (Table 2) we have some following remarks: Specifically, alginate concentration had main effect 165.2
and P value 0.00 then added cell had main effect 31.65 and P value 0.002. The analyzing data demonstrated R-
sq = 98.63% and Lack of fit test having p-value of 0.05, this meant that the immobilization efficiency and two
main effective factors were not in the linear relationship.
Table 2: The affecting level of two factors
Name of
factor
Symbol Value of factor Main effect P
valueLow (-1) Central (0) High (+1)
Alginate
concentration
(%)
X1 1.5 2.5 3.5 165.02 0.000
Cell added
(million
colonies/ml
X5 10 55 100 31.65 0.002
We identified the function of the poly-nominal regression accurately. The experiment for response surface
methodology having the central composite designs (RMS-CCD) with the main effective factors (-α, -1, 0. +1
+α) were conducted through 13 experiments, including 5 central experiments, 4 corner experiments and 4 axial
experiments.
The highest cell immobilization efficiency was the 4th
run (X1=0, X5= -α) of all experiments, which reached
90.73%. The lowest cell immobilization efficiency was the 3rd
run (X1= -1, X5= 1) which reached 30.23%, the
efficiency of immobilization. The experiments were illustrated in Table 3.
Table 3: The efficiency of immobilization Bacillus subtilis natto by alginate – chitosan complex in central
composite designs.
Run Code value Un code value Efficiency
Y (%)X1 X5 X1
(%)
X5
(million colony/ ml)
1 0 0 2.5 55 75.67
2 -1 -1 1,5 10 39.76
3 -1 1 1.5 100 30.23
4 0 -1.41 2.5 5.858 90.73
5 0 0 2,5 55 74,01
6 -1.41 0 1.085 55 32,41
7 1.41 0 3.914 55 45.56
8 0 0 2.5 55 76.58
9 1 1 3.5 100 59.32
10 1 -1 3.5 10 80.27
11 0 0 2.5 55 75.85
12 0 +1.41 2.5 104.142 42.71
13 0 0 2.5 55 76.36
R-sq – 90.15%
In this case, we determined the relationship between the main effective factors and response. The poly-nominal
regression equation was determined below.
Y (%) = -67.86 + 106.86*X1 + 1.35*X5 – 18.47*X1
2
- 2.27*X5
2
– 6.34*X1*X5
In above equation, Y (%) is symbol for cell immobilization efficiency, X1 (%) is symbol for alginate
concentration, X5 (million colonies per milliliter) is symbol for added cell density.
4. Optimization Of Bacillus Subtilis Natto Immobilization Process On Alginate – Chitosan Complex..
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Figure 1: The equation contours show the relationship between Y (%) and X1, X5
When added cells density was above central value, immobilization efficiency was reduced. This shows that,
added cells density being higher than central value will lead to competitive nutrients of cells in the gel bead.
Besides, the increasing by-product in the fermentation medium leads to inhibition of cell grows. Therefore, cells
enter death phase and the cells density in the gel bead decreases.
When alginate concentration increased above the central value, the immobilization efficiency also increased.
However, when alginate concentration reached value +1, despite the increasing in alginate of concentration the
immobilization efficiency was not increased.
The reason is that when alginate concentration increases, the viscosity increases too, so the ability of diffusing
cells into alginate suspensions is limited. On the other hand, when alginate concentration decreases below the
central value, the immobilization efficiency also decreases because low alginate concentration lead to the large
size pore gel, cells in gel bead are easily released to fermentation medium.
The maximum efficiency of immobilization Bacillus subtilis natto on alginate – chitosan complex was 90.73%
when alginate concentration is 2.5% and added cell density is 8.58 million colonies per milliliter, the others
factors remaining at central level.
1.5 Using immobilization Bacillus subtilis natto for fermentation of nattokinase
1.5.1 Investigating biosynthesis process of nattokinase enzyme based on time of fermentation.
Survey time period started at 8th
hour and stopped at 28th
hour of fermentation. The nattokinase enzyme activity
obtained increased as survey time increased. At the time of 8th
hour after fermentation, nattokinase enzyme
activity reached 31.46 FU/ml because in this stage, cells was in log phase and using nutrition for growing. The
results of survey process were illustrated in the figure 2.
Figure 2: The effect of fermenting time on biosynthesis of nattokinase enzyme process.
After 20 hours of fermentation, nattokinase enzyme activity reached 67.21 FU/ml and increased 2.2 fold
compared with at the 8th
hour of fermentation. The time from 24th
hour to 28th
hour correspond with the time
when cells in the stationary phase so nattokinase enzyme activity just slightly increased by 0.16 FU/ml. This
indicated that the increase in density of cells in the fermenting medium is proportional with obtained nattokinase
enzyme activity. At 28th
hour of fermentation nattokinase enzyme activity reached maximum 71.96 FU/ml.
However for the 4 hours prolonged time, nattokinase enzyme activity only increased 0.16 FU/ml having no
economic effect. Because the purpose of fermenting nattokinase enzyme by immobilized cells is to obtain the
highest enzyme activity and cell density, we chose the time to stop fermentation at the 24th
hour.
1.5.2 Comparing nattokinase enzyme activity obtained from fermenting process using free cells and
fermenting process using immobilized cells.
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From the investigating results obtained from biosynthesis of nattokinase enzyme process based on fermentation
time, we conducted fermenting immobilized cells and fermenting free cells with the same conditions. The result
was illustrated in figure 3.
Figure 3: Comparing nattokinase enzyme activity value obtaining from fermenting free cells and fermenting
immobilized cells.
For the free cells experiment, the maximum nattokinase enzyme activity obtained at 20th
hour of fermentation
was 69.13 FU/ml. Meanwhile, the maximum nattokinase enzyme activity obtained from fermenting the
immobilized cells was 71.96 FU/ml at 28th
hour of fermentation. The cause of the difference is that immobilized
cells take longer time to adapt with fermentation medium than free cells do. With free cell, when cells were
added in to fermentation broth, they immediately and directly contacted with the substrate. As for the
immobilized cells, cells were locked up behind the barrier layer of particle gel so cells used substrate after being
diffused through the membrane. On the other hand, the diffusion also has the limitation because it is effected by
concentration of substrate and size of the substrate. However for free cell, when increase time for fermented
from 20th
hour to 28th
hour, the nattokinase enzyme activity obtained was not increase. While the nattokinase
enzyme activity obtained by the immobilized cell was increased 4.75 FU/ml.
1.5.3 Investigating nattokinase enzyme activity in a series of immobilized cells reuse
We conducted survey nattokinase enzyme activity for a number of immobilized cells reuse times. In every 24
hours, we stopped the fermentation, transferred gel beads to fresh broth and continued fermenting process with
the same condition on the fresh broth. The results of investigating nattokinase enzyme activity over time of
reuse were illustrated in the figure 4.
Figure 4: Survey reusing the immobilized cells
The nattokinase enzyme activity in the first reuse reached 73.13 FU/ml, the highest level among the reuse times
and higher than the initial of immobilized cells. Particularly, the nattokinase enzyme activity obtained in the first
reuse and the second reuse were higher than the initial of immobilized cells by 1.33 FU/ml and 0.61 FU/ml
respectively. The cause of this difference is due to when we stopped process of fermented at the time of 24th
hour, the cells in gel beads are ungrowing hormogenous. Some are growing, others might be in the process of
biosynthesis of nattokinase enzyme thus nattokinase enzyme activity obtained was not in the highest level at the
initial fermentation of the immobilized cells. At the time of the first reuse nattokinase enzyme activity obtained
6. Optimization Of Bacillus Subtilis Natto Immobilization Process On Alginate – Chitosan Complex..
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reached the highest level because cells in the gel bead are not undergo growth stages which mainly using
nutrition for the biosynthesis of nattokinase enzyme process. At the next time reuses, obtained nattokinase
enzyme activity decreasing compared with the previous reuses may be due to after each reuse, a number of cells
were being released in to fermentation broth. So cells density in the gel beads were decreased and obtained
nattokinase enzyme activity were also decreased.
However after reusing for 6 times, obtained nattokinase enzyme activity compared with the 1st
time of reuse is
decrease by 2.73 FU/ml, this reduction is not significant. The survey result suggests that alginate – chitosan
complex had its effect in reducing cells release and increasing times of reuse.
IV. Conclusion
By the response surface methodology, we had investigated two factors affecting the immobilization efficiency
are: alginate concentration and added cell density. The Optimized of parameters for six factors are concentration
of alginate (2.5%), concentration of chitosan (0.4%), pH of chitosan (5.5), concentration of CaCl2 (2%), added
cell density (approximately 8.58 colonies per milliliter) and shaking time when supplement chitosan (30
minutes). The highest of immobilization efficiency reached 90.73%.
The immobilized cells were reused 6 times and obtained nattokinase enzyme activity in the 1st
time of reuse and
in the 6th
of reuse are 73.13 ± 0.14 FU/ml and 70.40 ± 0.09 FU/ml respectively.
Acknowledgment
We would like to full appreciate the help of the professors/teachers in the Chemistry Faculty of Ho Chi Minh
University of technology in this study. This review is funded by Vietnam National University Ho Ch iMinh City
(VNU-HCM) under grant number C2016-20-31.
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