Optimization of conditions for cholesterol oxidase production by the microorganism isolated from urban compost and dairy soil samples.Isolates were obtained on the basis of their capability of growing on isolation medium A and B and their cholesterol oxidase (CHO) production was estimated. CHO production was optimized by the optimization of temperature, pH, carbon sources, and organic and inorganic nitrogen sources.isolates out of 22 were found to secrete extracellular CHO as detected by cholesterol oxidase indicator plate A and were designated as cholesterol oxidase producing isolate 1, 2 and 3 (COP 1, COP 2 and COP 3). Results showed that the strain COP 2 belonging to the genus Rhodococcus sp. based on morphological, cultural and biochemical characteristics recorded highest cholesterol oxidase activity. Optimum temperature and pH for CHO activity were found to be 35 °C and 7.5 respectively. Steroidal substrate cholesterol produced a significant increase in CHO level (0.502 IU/ml). Organic and inorganic nitrogen sources were supplemented in combinations leads to increase in CHO production as compared to individual components.
Isolation of Yeasts from Raisins and Palm-Juice and Ethanol Production in Mol...Shafkat Shamim Rahman
The alternative fuels are expected to satisfy the progressive demand for energy on the wake of the negative effects of fossil fuel on the atmosphere and resultant universal warming. In this study two ethanol fermenting Saccharomyces cerevisae were isolated from Palm juice and Raisins. Both isolates were grown in Yeast extract Peptone Dextrose (YEPD) medium and characterized for alcoholic fermentation using molasses medium and optimized for pH, thermo-, osmo-, ethanol tolerance and sugar concentration. Results showed for ethanol fermentation, 31°C temperature, 6.01 pH and 6.50% sugar concentration is the prime condition. Raisin-isolate emerged as highly thermophilic and stress tolerant in nature. Under optimized conditions, S. cerevisae isolated from Palmjuice produced 9.85% of ethanol in the medium. Creation of ethanol through fermentation appears to be a potential other fossil fuel and can be used as exclusive fuel in vehicles with dedicated engines or in fuel blends.
Isolation of Saccharomyces cerevisiae from pineapple and orange and study of ...Shafkat Shamim Rahman
Poster presented at National Conference 2016 on “Biochemistry and Molecular Biology for Life Sciences”,
Organized by:
The Bangladesh Society for Biochemistry and Molecular Biology in association with Department of Biochemistry and Molecular Biology, University of Dhaka,
Held at December 10, 2016, Nabab Nawab Ali Chowdhury Senate Bhavan, University of Dhaka, Dhaka, Bangladesh.
This document summarizes a study that screened eight Monascus strains for their ability to produce lovastatin, an alternative to statin drugs. Strain M. purpureus Went (JCM 6934) produced the highest amount of lovastatin at 84.85 ppm. Angkak rice fermented with the local Philippine strain M. purpureus UPLB-MNH-MCC 2108 was found to also contain lovastatin. This showed that lovastatin is a characteristic component of angkak rice. A preliminary study found very low levels of the toxin citrinin in cookies made with angkak rice, showing it is safe for human consumption.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
Flavoring and medicinal values of the yellow pigment produced by Monascus rub...Premier Publishers
The Thin Layer Chromatographic analysis (TLC) of the crud red pigment extract of M. ruber 4066 cultivated on the malt static medium well developed into three separate bands includes red, orange and yellow pigmented bands. Sixty four volatile metabolites are detected by GC/MS analysis which gave yellow pigment high flavoring value. The detected metabolites are classified, chemical and physical properties are characterized, and their uses are reported. The detected metabolites including fourteen aliphatic metabolites, twenty nine aromatic metabolites and terpenoids, thirteen nitrogenous metabolites and also five heterocyclic nitrogenous thiols metabolites are detected. Also they are classified into thirteen volatile and aroma groups includes one metabolite from the each "alcohol, acid, aldehyde, amine, furan and mercapto", two phenols, three amides, four from each "indoles and ketones", twelve terpenoids, fourteen alkanes & alkenes, sixteen esters and three metabolites are unknown for me.
This document summarizes research on producing penicillin through solid state fermentation using sugarcane bagasse as a support medium. Key findings include:
1) Using a concentrated culture medium (2X) greatly enhanced penicillin production in solid state fermentation compared to liquid fermentation or using non-concentrated medium.
2) An initial moisture content of 70% of the impregnated solid medium resulted in higher penicillin production and stability than lower or higher moisture contents.
3) Solid state fermentation yielded significantly higher penicillin production (17 times), faster production time (one third the time), and greater efficiency than liquid fermentation using the same microorganism and medium.
Isolation of Yeasts from Raisins and Palm-Juice and Ethanol Production in Mol...Shafkat Shamim Rahman
The alternative fuels are expected to satisfy the progressive demand for energy on the wake of the negative effects of fossil fuel on the atmosphere and resultant universal warming. In this study two ethanol fermenting Saccharomyces cerevisae were isolated from Palm juice and Raisins. Both isolates were grown in Yeast extract Peptone Dextrose (YEPD) medium and characterized for alcoholic fermentation using molasses medium and optimized for pH, thermo-, osmo-, ethanol tolerance and sugar concentration. Results showed for ethanol fermentation, 31°C temperature, 6.01 pH and 6.50% sugar concentration is the prime condition. Raisin-isolate emerged as highly thermophilic and stress tolerant in nature. Under optimized conditions, S. cerevisae isolated from Palmjuice produced 9.85% of ethanol in the medium. Creation of ethanol through fermentation appears to be a potential other fossil fuel and can be used as exclusive fuel in vehicles with dedicated engines or in fuel blends.
Isolation of Saccharomyces cerevisiae from pineapple and orange and study of ...Shafkat Shamim Rahman
Poster presented at National Conference 2016 on “Biochemistry and Molecular Biology for Life Sciences”,
Organized by:
The Bangladesh Society for Biochemistry and Molecular Biology in association with Department of Biochemistry and Molecular Biology, University of Dhaka,
Held at December 10, 2016, Nabab Nawab Ali Chowdhury Senate Bhavan, University of Dhaka, Dhaka, Bangladesh.
This document summarizes a study that screened eight Monascus strains for their ability to produce lovastatin, an alternative to statin drugs. Strain M. purpureus Went (JCM 6934) produced the highest amount of lovastatin at 84.85 ppm. Angkak rice fermented with the local Philippine strain M. purpureus UPLB-MNH-MCC 2108 was found to also contain lovastatin. This showed that lovastatin is a characteristic component of angkak rice. A preliminary study found very low levels of the toxin citrinin in cookies made with angkak rice, showing it is safe for human consumption.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
Flavoring and medicinal values of the yellow pigment produced by Monascus rub...Premier Publishers
The Thin Layer Chromatographic analysis (TLC) of the crud red pigment extract of M. ruber 4066 cultivated on the malt static medium well developed into three separate bands includes red, orange and yellow pigmented bands. Sixty four volatile metabolites are detected by GC/MS analysis which gave yellow pigment high flavoring value. The detected metabolites are classified, chemical and physical properties are characterized, and their uses are reported. The detected metabolites including fourteen aliphatic metabolites, twenty nine aromatic metabolites and terpenoids, thirteen nitrogenous metabolites and also five heterocyclic nitrogenous thiols metabolites are detected. Also they are classified into thirteen volatile and aroma groups includes one metabolite from the each "alcohol, acid, aldehyde, amine, furan and mercapto", two phenols, three amides, four from each "indoles and ketones", twelve terpenoids, fourteen alkanes & alkenes, sixteen esters and three metabolites are unknown for me.
This document summarizes research on producing penicillin through solid state fermentation using sugarcane bagasse as a support medium. Key findings include:
1) Using a concentrated culture medium (2X) greatly enhanced penicillin production in solid state fermentation compared to liquid fermentation or using non-concentrated medium.
2) An initial moisture content of 70% of the impregnated solid medium resulted in higher penicillin production and stability than lower or higher moisture contents.
3) Solid state fermentation yielded significantly higher penicillin production (17 times), faster production time (one third the time), and greater efficiency than liquid fermentation using the same microorganism and medium.
The study analyzed 48 pesticides including organochlorines, organophosphates, synthetic pyrethroids, and herbicides in 60 samples of 20 vegetables collected from Lucknow, India using the QuEChERS extraction method and gas chromatography. 23 pesticides were detected in the vegetable samples at levels ranging from 0.005-12.35 mg/kg. Some vegetables like radish, cucumber, cauliflower, cabbage, and okra contained pesticide residues above the Indian maximum residue limits for pesticides in foods. The study assessed the pesticide contamination of vegetables commonly consumed in Lucknow.
A comparative study of In-vitro Thrombolytic activity and Anti-inflammatory a...Anindya_shuvo773
The document presents research on the thrombolytic and anti-inflammatory properties of methanolic leaf extracts of Blumea lacera and Acanthus ilicifolius. In vitro assays found that the B. lacera extract had 49.27% clot lysis compared to 38.25% for A. ilicifolius, with streptokinase as the positive control at 69.34%. Anti-inflammatory assays showed the B. lacera extract inhibited protein denaturation by 64.56% versus 45.27% for A. ilicifolius, compared to the standard acetyl salicylic acid. Overall, the results indicate the leaf extracts of both plants show significant thrombolytic and anti-
This document summarizes a study on the effects of environmental factors on fungal α-amylase production using cereal processing mill residues as substrates. Key findings include:
- An Aspergillus sp. strain isolated from soil samples showed high amylase production. Wheat bran supported maximum enzyme production among various residues tested.
- Maximum amylase activity was achieved under optimized environmental factors - 60% initial moisture, pH 5, incubation at 30°C, 4ml inoculum volume, and 20g substrate in 500ml flask.
- One-factor-at-a-time experiments revealed initial moisture content, temperature, pH, inoculum level, and substrate-volume ratio significantly influence enzyme yield during
The document examines the peroxidase activity in arracacha (Arracacia xanthorrhyza Bancroft) roots affected by pH and temperature. Roots were stored at 5°C to induce chilling injury and then peroxidase kinetic activity was determined under different pH and temperature conditions. The peroxidase activity was highest between pH 5.5-6.0 and at 30°C. The enzyme was more susceptible to inactivation at acidic pH levels compared to alkaline pH levels and could also be inactivated through heat treatment.
Effect of Various Parameters on the Growth and Ethanol Production by Yeasts I...Shafkat Shamim Rahman
Two ethanol fermenting Saccharomyces cerevisiae were isolated from date juice and grapes and grown in YEPD medium. They were characterized for alcoholic fermentation using sugarcane molasses and their growth conditions were optimized with respect to pH and sugar concentration. Results revealed a temperature of 30ºC, pH 6.0 and 6.5% sugar concentration as optimum for fermentation. Stress tolerance tests showed that date juice isolate was highly tolerant to temperature, pH and high ethanol concentration in the medium. Under optimized conditions, S. cerevisiae isolated from date-juice produced 7.75% of ethanol in molasses as estimated by Conway method.
The endo-glucanase (E.C. 3.2.1.4) was produced by Aspergillus terreus adopting solid state fermentation (SSF) using agro residues as main substrate. To recover the enzyme from the fermented mass, different extraction liquids were tried and 10% aqueous solution of glycerol was found to be superior. When the selected extractant was applied at different ratio to the fermented solid mass, maximum enzyme was recovered at 1:5 (w/v) ratio. The other process parameters (time, temperature and mixing speed) effects on the enzyme recovery were subsequently studied by response surface methodology (RSM). Box-Bhenken Design of experiment
Taylor created XL (Xylose Lysine) Agar Base to isolate and differentiate Gram-negative enteric bacteria.
Sodium thiosulfate, ferric ammonium citrate, and sodium deoxycholate were added to XL Agar Base to create XLD Agar, a more selective medium.
Using numerous staining chemicals, George Chapman and his colleagues at The Clinical Research Laboratory in New York produced a series of isolation media in the 1930s and 1940s.
A yeast strain E2 was purified from traditional yeast, and retained for its strongly acidifying, fermentative and saccharolytic
power. In fact, this strain produces a high concentration of acetic acid 105.85 mg / L revealed by using the H.P.L.C DAD technique
during its growth in semi synthetic medium containing sucrose at 5 g /l as only carbon source. The pH of the culture medium increases
from 5.58 to 2.76 after 24 hours of culture and to 2.48 after 48 hours of
Isolation and analytical characterization of local malaysian leech saliva ext...Younis I Munshi
This study isolated and characterized proteins and peptides from the saliva of local Malaysian leeches. A modified extraction method was used to obtain leech saliva without harming the leeches. The saliva extract was found to contain high concentrations of proteins using UV and Bradford assays. Gel electrophoresis revealed over 25 protein and peptide bands between 3.7-80 kDa. RP-HPLC showed over 30 distinct peaks in the saliva extract. Some proteins matched known proteins from related leech species, while others were potentially novel biologically active compounds. This research provides a basis for further investigation of proteins in Malaysian leech saliva that may have applications in medicine.
The document summarizes a microbiology lab exercise on carbohydrate metabolism. Students will check results from the previous nitrogen metabolism lab, inoculate carbohydrate metabolism samples, and perform experiments including: fermentation tube tests of different carbohydrates; detection of fermentation products using MRVP media; testing for starch hydrolysis; and determining citrate utilization. Key concepts covered include: carbohydrates as preferred carbon sources; fermentation pathways and end products; and use of different media like Kligler Iron Agar, starch agar, and Simmon's Citrate Agar to analyze microbial metabolism and unknown bacteria.
Isolation and characterization of lipolytic Pseudomonas spp. from oil contami...iosrjce
Oil contaminated water samples collected from different areas in Pune were screened for the
selective isolation of lipolytic Pseudomonas spp. Screening medium containing Tributyrin (1% v/v) and
Rhodamine B was used for the specific isolation of lipolytic Pseudomonas spp. from oil contaminated water
samples. Lipolytic bacteria showing zone of clearance around the colonies were selected for the present study.
Isolated bacteria were identified upto genus level with the help of morphological and biochemical testing by
following Bergey’s manual. Spectrophotometric Lipase assay showed that out of 49 lipolytic Pseudomonas spp.,
WP23 exhibits the maximum Lipase activity of about 45.4733 U/ml.
An Investigation Into The Mechanisms Underlying Enhanced Biosulphidogenesis I...iosrjce
Anthropogenic activities like mining, processes of metallurgy and other chemical industries lead to
the discharge of a high amount of sulphate into the environment that causes serious problems to human health.
This paper illustrates the employment of thermophilic sulphate reducing bacteria for biosulphidogenesis. Two
different species have been isolated from hot water spring of Vajreshwari and Ganeshpuri,Thane, Maharashtra,
INDIA.The mechanism involved in biosulphidogenesis includes production of specific protein as well as
liberation of some extracellular polymeric compound (EPS) e.g. proteins, carbohydrate, acids etc. that are
produced during the microbial cell metabolism. These compounds plays an important role in the faster
reduction of sulphate and decrease in production rate of sulphide.The isolate was found to be of genus
Bacillusand type strain was found to be subtilis Zankar and licheniformis Sonali. The strain sequence were
deposited in NCBI database with accession number KJ939324 and KJ939325 respectively. The result highlights
the potential use of these organism in biosulphidogenesis.
Asparaginase is an important enzyme in Medicine & food industry. It catalyzes Asparagine to aspartate and Ammonia. The purpose of using asparaginase in foods is to reduce the levels of acrylamide that form in certain carbohydrate-rich foods during cooking.The rationale behind asparaginase is that it takes advantage of the fact that acute lymphoblastic leukemia cells and some other suspected tumor cells are unable to synthesize the non-essential amino acid asparagine, whereas normal cells are able to make their own asparagine.
This document discusses enzymes called asparaginase. It begins by explaining that enzymes are proteins that act as biological catalysts. It then discusses hydrolases, a class of enzymes that catalyze the hydrolysis of chemical bonds. Asparaginase is introduced as a commercially important hydrolase. The document provides information on the sources, mechanism of action, and use of asparaginase as a food processing aid to reduce acrylamide formation. It then describes the materials and methods used for asparaginase production, including media preparation, isolation, screening, and analysis of enzyme activity with respect to temperature and pH variations.
This study was conducted to study the effects of supplementation alfalfa silage with orange pulp and difference of Lactobacillus buchneri on in vitro dry matter digestibility and gas production. wilted alfalfa with no additive (control), wilted Alfalfa and orange pulp (1750 g wilted Alfalfa mixed with 750 g fresh orange pulp) treated with LAB for final application rates of 0, 2.5, 5 and 7.5 g LAB inoculant/ton of wilted alfalfa and orange pulp (LAB0, LAB1, LAB2, LAB3, respectively). Alfalfa hay harvested at flowering stage and after 24 hours wilted and mixed orange pomace with ratio of 2100 g and 760 g, respectively, and was ensiled for 90 days. The data were analyzed in a completely randomized design with three replications. After 24 h incubation, treatments AO (alfalfa + orange pulp) and CON (without additive) had the highest and lowest in vitro gas production (p<0.05) and adding orange pulp and molasses increased gas production. Adding inoculant decreased in vitro DM digestibility. Results showed that ensiling alfalfa with orange pulp and molasses can improved silage quality and increased gas production and in vitro DM digestibility.
This document summarizes a study on the fermentative production of squalene using Saccharomyces cerevisiae and Torulaspora delbrueckii under anaerobic conditions. Squalene yields of 41.16 μg/g and 237.25 μg/g were obtained from S. cerevisiae and T. delbrueckii, respectively. Isolation and purification of squalene from lipid extracts of the yeast cells was achieved through chromatographic techniques. Spectroscopic characterization confirmed the purified product was squalene.
The document is a training report submitted by Sapna Singh to Sam Higginbottom Institute of Agriculture, Technology and Sciences. It discusses 16 experiments conducted on advanced biotech techniques under the guidance of Dr. Vineeta Singh at MRD LifeSciences. The experiments included isolation of bacteria from soil, purification of cultures, screening for amylase production, studying bacterial growth curves, and enzyme assays. acknowledgements are provided to various individuals and organizations that supported the training.
1. The document discusses the topics of fermentation technology and processes. It covers definitions of fermentation, important fermentation products, fermenter types, medium composition, inoculation, and microbial rates and stoichiometry.
2. Key aspects of fermentation covered include the use of stirred tank bioreactors, types of fermenters including batch, fed-batch and continuous reactors, and important industrial fermentation products like ethanol, lactic acid, and antibiotics.
3. Microbial rates are quantified as specific rates and yields, and stoichiometric equations are used to represent microbial growth coupling catabolism and anabolism based on substrate utilization and energy generation.
This document summarizes restrictions on sharing and distributing an article from a journal published by Elsevier. The copy is for the author's personal non-commercial use, including instruction and sharing with colleagues. Other uses like reproduction, distribution, selling, licensing copies or posting on websites are prohibited without permission. Authors can generally post their version of the article to their personal or institutional websites or repositories. The document provides a link for more information on Elsevier's archiving and manuscript policies.
This document summarizes restrictions on sharing and distributing an article from a journal published by Elsevier. The attached copy can be used by the author for non-commercial research and education purposes, including instruction and sharing with colleagues, but other uses like reproduction, distribution, selling, licensing copies or posting to third party websites are prohibited without permission. The author is allowed to post a version of the article to their personal website or institutional repository in most cases. Further information on Elsevier's archiving and manuscript policies is available at the provided URL.
The study analyzed 48 pesticides including organochlorines, organophosphates, synthetic pyrethroids, and herbicides in 60 samples of 20 vegetables collected from Lucknow, India using the QuEChERS extraction method and gas chromatography. 23 pesticides were detected in the vegetable samples at levels ranging from 0.005-12.35 mg/kg. Some vegetables like radish, cucumber, cauliflower, cabbage, and okra contained pesticide residues above the Indian maximum residue limits for pesticides in foods. The study assessed the pesticide contamination of vegetables commonly consumed in Lucknow.
A comparative study of In-vitro Thrombolytic activity and Anti-inflammatory a...Anindya_shuvo773
The document presents research on the thrombolytic and anti-inflammatory properties of methanolic leaf extracts of Blumea lacera and Acanthus ilicifolius. In vitro assays found that the B. lacera extract had 49.27% clot lysis compared to 38.25% for A. ilicifolius, with streptokinase as the positive control at 69.34%. Anti-inflammatory assays showed the B. lacera extract inhibited protein denaturation by 64.56% versus 45.27% for A. ilicifolius, compared to the standard acetyl salicylic acid. Overall, the results indicate the leaf extracts of both plants show significant thrombolytic and anti-
This document summarizes a study on the effects of environmental factors on fungal α-amylase production using cereal processing mill residues as substrates. Key findings include:
- An Aspergillus sp. strain isolated from soil samples showed high amylase production. Wheat bran supported maximum enzyme production among various residues tested.
- Maximum amylase activity was achieved under optimized environmental factors - 60% initial moisture, pH 5, incubation at 30°C, 4ml inoculum volume, and 20g substrate in 500ml flask.
- One-factor-at-a-time experiments revealed initial moisture content, temperature, pH, inoculum level, and substrate-volume ratio significantly influence enzyme yield during
The document examines the peroxidase activity in arracacha (Arracacia xanthorrhyza Bancroft) roots affected by pH and temperature. Roots were stored at 5°C to induce chilling injury and then peroxidase kinetic activity was determined under different pH and temperature conditions. The peroxidase activity was highest between pH 5.5-6.0 and at 30°C. The enzyme was more susceptible to inactivation at acidic pH levels compared to alkaline pH levels and could also be inactivated through heat treatment.
Effect of Various Parameters on the Growth and Ethanol Production by Yeasts I...Shafkat Shamim Rahman
Two ethanol fermenting Saccharomyces cerevisiae were isolated from date juice and grapes and grown in YEPD medium. They were characterized for alcoholic fermentation using sugarcane molasses and their growth conditions were optimized with respect to pH and sugar concentration. Results revealed a temperature of 30ºC, pH 6.0 and 6.5% sugar concentration as optimum for fermentation. Stress tolerance tests showed that date juice isolate was highly tolerant to temperature, pH and high ethanol concentration in the medium. Under optimized conditions, S. cerevisiae isolated from date-juice produced 7.75% of ethanol in molasses as estimated by Conway method.
The endo-glucanase (E.C. 3.2.1.4) was produced by Aspergillus terreus adopting solid state fermentation (SSF) using agro residues as main substrate. To recover the enzyme from the fermented mass, different extraction liquids were tried and 10% aqueous solution of glycerol was found to be superior. When the selected extractant was applied at different ratio to the fermented solid mass, maximum enzyme was recovered at 1:5 (w/v) ratio. The other process parameters (time, temperature and mixing speed) effects on the enzyme recovery were subsequently studied by response surface methodology (RSM). Box-Bhenken Design of experiment
Taylor created XL (Xylose Lysine) Agar Base to isolate and differentiate Gram-negative enteric bacteria.
Sodium thiosulfate, ferric ammonium citrate, and sodium deoxycholate were added to XL Agar Base to create XLD Agar, a more selective medium.
Using numerous staining chemicals, George Chapman and his colleagues at The Clinical Research Laboratory in New York produced a series of isolation media in the 1930s and 1940s.
A yeast strain E2 was purified from traditional yeast, and retained for its strongly acidifying, fermentative and saccharolytic
power. In fact, this strain produces a high concentration of acetic acid 105.85 mg / L revealed by using the H.P.L.C DAD technique
during its growth in semi synthetic medium containing sucrose at 5 g /l as only carbon source. The pH of the culture medium increases
from 5.58 to 2.76 after 24 hours of culture and to 2.48 after 48 hours of
Isolation and analytical characterization of local malaysian leech saliva ext...Younis I Munshi
This study isolated and characterized proteins and peptides from the saliva of local Malaysian leeches. A modified extraction method was used to obtain leech saliva without harming the leeches. The saliva extract was found to contain high concentrations of proteins using UV and Bradford assays. Gel electrophoresis revealed over 25 protein and peptide bands between 3.7-80 kDa. RP-HPLC showed over 30 distinct peaks in the saliva extract. Some proteins matched known proteins from related leech species, while others were potentially novel biologically active compounds. This research provides a basis for further investigation of proteins in Malaysian leech saliva that may have applications in medicine.
The document summarizes a microbiology lab exercise on carbohydrate metabolism. Students will check results from the previous nitrogen metabolism lab, inoculate carbohydrate metabolism samples, and perform experiments including: fermentation tube tests of different carbohydrates; detection of fermentation products using MRVP media; testing for starch hydrolysis; and determining citrate utilization. Key concepts covered include: carbohydrates as preferred carbon sources; fermentation pathways and end products; and use of different media like Kligler Iron Agar, starch agar, and Simmon's Citrate Agar to analyze microbial metabolism and unknown bacteria.
Isolation and characterization of lipolytic Pseudomonas spp. from oil contami...iosrjce
Oil contaminated water samples collected from different areas in Pune were screened for the
selective isolation of lipolytic Pseudomonas spp. Screening medium containing Tributyrin (1% v/v) and
Rhodamine B was used for the specific isolation of lipolytic Pseudomonas spp. from oil contaminated water
samples. Lipolytic bacteria showing zone of clearance around the colonies were selected for the present study.
Isolated bacteria were identified upto genus level with the help of morphological and biochemical testing by
following Bergey’s manual. Spectrophotometric Lipase assay showed that out of 49 lipolytic Pseudomonas spp.,
WP23 exhibits the maximum Lipase activity of about 45.4733 U/ml.
An Investigation Into The Mechanisms Underlying Enhanced Biosulphidogenesis I...iosrjce
Anthropogenic activities like mining, processes of metallurgy and other chemical industries lead to
the discharge of a high amount of sulphate into the environment that causes serious problems to human health.
This paper illustrates the employment of thermophilic sulphate reducing bacteria for biosulphidogenesis. Two
different species have been isolated from hot water spring of Vajreshwari and Ganeshpuri,Thane, Maharashtra,
INDIA.The mechanism involved in biosulphidogenesis includes production of specific protein as well as
liberation of some extracellular polymeric compound (EPS) e.g. proteins, carbohydrate, acids etc. that are
produced during the microbial cell metabolism. These compounds plays an important role in the faster
reduction of sulphate and decrease in production rate of sulphide.The isolate was found to be of genus
Bacillusand type strain was found to be subtilis Zankar and licheniformis Sonali. The strain sequence were
deposited in NCBI database with accession number KJ939324 and KJ939325 respectively. The result highlights
the potential use of these organism in biosulphidogenesis.
Asparaginase is an important enzyme in Medicine & food industry. It catalyzes Asparagine to aspartate and Ammonia. The purpose of using asparaginase in foods is to reduce the levels of acrylamide that form in certain carbohydrate-rich foods during cooking.The rationale behind asparaginase is that it takes advantage of the fact that acute lymphoblastic leukemia cells and some other suspected tumor cells are unable to synthesize the non-essential amino acid asparagine, whereas normal cells are able to make their own asparagine.
This document discusses enzymes called asparaginase. It begins by explaining that enzymes are proteins that act as biological catalysts. It then discusses hydrolases, a class of enzymes that catalyze the hydrolysis of chemical bonds. Asparaginase is introduced as a commercially important hydrolase. The document provides information on the sources, mechanism of action, and use of asparaginase as a food processing aid to reduce acrylamide formation. It then describes the materials and methods used for asparaginase production, including media preparation, isolation, screening, and analysis of enzyme activity with respect to temperature and pH variations.
This study was conducted to study the effects of supplementation alfalfa silage with orange pulp and difference of Lactobacillus buchneri on in vitro dry matter digestibility and gas production. wilted alfalfa with no additive (control), wilted Alfalfa and orange pulp (1750 g wilted Alfalfa mixed with 750 g fresh orange pulp) treated with LAB for final application rates of 0, 2.5, 5 and 7.5 g LAB inoculant/ton of wilted alfalfa and orange pulp (LAB0, LAB1, LAB2, LAB3, respectively). Alfalfa hay harvested at flowering stage and after 24 hours wilted and mixed orange pomace with ratio of 2100 g and 760 g, respectively, and was ensiled for 90 days. The data were analyzed in a completely randomized design with three replications. After 24 h incubation, treatments AO (alfalfa + orange pulp) and CON (without additive) had the highest and lowest in vitro gas production (p<0.05) and adding orange pulp and molasses increased gas production. Adding inoculant decreased in vitro DM digestibility. Results showed that ensiling alfalfa with orange pulp and molasses can improved silage quality and increased gas production and in vitro DM digestibility.
This document summarizes a study on the fermentative production of squalene using Saccharomyces cerevisiae and Torulaspora delbrueckii under anaerobic conditions. Squalene yields of 41.16 μg/g and 237.25 μg/g were obtained from S. cerevisiae and T. delbrueckii, respectively. Isolation and purification of squalene from lipid extracts of the yeast cells was achieved through chromatographic techniques. Spectroscopic characterization confirmed the purified product was squalene.
The document is a training report submitted by Sapna Singh to Sam Higginbottom Institute of Agriculture, Technology and Sciences. It discusses 16 experiments conducted on advanced biotech techniques under the guidance of Dr. Vineeta Singh at MRD LifeSciences. The experiments included isolation of bacteria from soil, purification of cultures, screening for amylase production, studying bacterial growth curves, and enzyme assays. acknowledgements are provided to various individuals and organizations that supported the training.
1. The document discusses the topics of fermentation technology and processes. It covers definitions of fermentation, important fermentation products, fermenter types, medium composition, inoculation, and microbial rates and stoichiometry.
2. Key aspects of fermentation covered include the use of stirred tank bioreactors, types of fermenters including batch, fed-batch and continuous reactors, and important industrial fermentation products like ethanol, lactic acid, and antibiotics.
3. Microbial rates are quantified as specific rates and yields, and stoichiometric equations are used to represent microbial growth coupling catabolism and anabolism based on substrate utilization and energy generation.
This document summarizes restrictions on sharing and distributing an article from a journal published by Elsevier. The copy is for the author's personal non-commercial use, including instruction and sharing with colleagues. Other uses like reproduction, distribution, selling, licensing copies or posting on websites are prohibited without permission. Authors can generally post their version of the article to their personal or institutional websites or repositories. The document provides a link for more information on Elsevier's archiving and manuscript policies.
This document summarizes restrictions on sharing and distributing an article from a journal published by Elsevier. The attached copy can be used by the author for non-commercial research and education purposes, including instruction and sharing with colleagues, but other uses like reproduction, distribution, selling, licensing copies or posting to third party websites are prohibited without permission. The author is allowed to post a version of the article to their personal website or institutional repository in most cases. Further information on Elsevier's archiving and manuscript policies is available at the provided URL.
Fungal cellulase xylanase production and corresponding hydrolysis using pretr...zhenhua82
Three pretreated corn stover (ammonia fiber expansion, dilute acid, and dilute alkali) were used as carbon source to culture Trichoderma reesei Rut C-30 for cellulase and xylanase production. The results indicated that the cultures on ammonia fiber expansion and alkali pretreated corn stover had better enzyme production than the acid pretreated ones. The consequent enzymatic hydrolysis was performed applying fungal enzymes on pretreated corn stover samples. Tukey’s statistical comparisons exhibited that there were significant differences on enzymatic hydrolysis among different combination of fungal enzymes and pretreated corn stover. The higher sugar yields were achieved by the enzymatic hydrolysis of dilute alkali pretreated corn stover.
The International Journal of Engineering and Science (The IJES)theijes
The International Journal of Engineering & Science is aimed at providing a platform for researchers, engineers, scientists, or educators to publish their original research results, to exchange new ideas, to disseminate information in innovative designs, engineering experiences and technological skills. It is also the Journal's objective to promote engineering and technology education. All papers submitted to the Journal will be blind peer-reviewed. Only original articles will be published.
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is an open access international journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Production of α-amylase using new strain of Bacillus polymyxa isolated from s...IOSR Journals
In this study, a new amylase producer strain was isolated from sweet potato tuber. This strain was able to grow at 37 °C and produce α-amylase in high quantity compared to other standard strain cultures. In the first part, cultivation in shake flask in standard medium was carried out to give complete information about the growth and production kinetics of this strain. The results clearly demonstrate that the isolated strain is able to production α-amylase in submerged culture with concentration up to 2050 kat/L after 20 h cultivation. Furthermore, medium optimization was carried out by changing the starch concentration and cell cultivation in medium of mixed carbon source (composed of starch and glucose of ratio 15:5 g/g) to enhance the production process and to increase the growth rate. The volumetric and specific α-amylase production in this optimized medium were 4550 kat/L and 1060 kat/g, respectively. Further improvement in enzyme production process was achieved by scaling up the process from shake flask to 3-L stirred tank bioreactor under non-oxygen limiting condition. The maximal volumetric and specific α-amylase productions in bioreactor batch culture were 5210 kat/L and 1095kat/g, respectively, after only 14 h cultivation
This document summarizes a study that investigated using molasses and corncob as alternative substrates for producing citric acid through fermentation with Aspergillus niger. The control medium containing sucrose produced 4.6 mg/ml of citric acid. The medium with molasses produced 10.4 mg/ml, while the medium with corncob produced 5.3 mg/ml. A medium with both molasses and 5% sucrose produced the highest yield of 12.6 mg/ml. Factors like incubation temperature, nitrogen supplements, and methanol concentration were optimized. The results show that molasses and corncob can be effective alternatives to sucrose for cost-effective citric acid production.
The Role of Cell Wall-Degrading Enzymes in the Development of Anthracnose Dis...Agriculture Journal IJOEAR
This document summarizes a study that evaluated the production of cell wall-degrading enzymes by Colletotrichum truncatum CP2, a fungal pathogen that causes anthracnose disease in chili peppers. The study found that polygalacturonase (PG) was the first cell wall-degrading enzyme detected, with higher activity levels than other enzymes. After PG degraded the cell wall, further degradation was carried out by pectin methylesterases, pectin lyase, and pectate lyase. C. truncatum CP2 produced higher levels of these enzymes compared to the reference fungus C. gloeosporiodes. The timing of peak enzymatic activity suggests each enzyme plays a specific
Plant growth promoting characterization of soil bacteria isolated from petrol...Agriculture Journal IJOEAR
Abstract— Contaminant-degrading bacteria can be included among the plant-growth promoting bacteria; because the presence of contaminants, in general produce negatively effects on plant’s growth; thus, the elimination of the inhibiting contaminants will benefit them. Although contaminant-degrading strains have been traditionally isolated from various environments; the number of studies that reported the isolation and identification of soil bacteria with contaminant- degrading abilities have increased. The aim of this study was to characterized microbial strains isolated from petroleum contaminated soil by plant growth promotion traits to recommend them as potential bioinoculants. In this work, five of the six soil isolates were classified as Indole Acetic Acid higher producers and only one of them as lower producer. Sporosarcina aquimarina strain -Q3 and Bacillus cereus strain +F2 tested in Axonopus affinis plantlets bioassay, showed that these isolates were the most effective promoters of this plant species; therefore, these soil bacteria with possible hydrocarbon degradation ability could be considered as potential bioinoculants and can be recommended with a practical importance for the rhizoremediation of petroleum contaminated sites and plant growth promotion.
1) The study investigated the polyphenolic content of rose hip (Rosa canina L.) tea extracts obtained using ultrasound-assisted extraction (UAE) and Soxhlet extraction with different solvents.
2) The highest extract yield was obtained using UAE with water at 619.37 mg/g dried matter. The highest total phenolic content was obtained using Soxhlet extraction with a 50% methanol mixture at 59.69 mg gallic acid equivalent/g dried matter.
3) UAE and Soxhlet extraction with various solvents, including water, ethanol, methanol, and mixtures, were tested on three commercial brands of rose hip tea. Total phenolic content and extract yields
Sensitive and selective detection of chemical residues in hops is necessary to ensure protection of consumers and the environment. Methods using LC-MS provide efficient and effective detection of chemical residues in a complex sample matrix such as hops. Presented here is an LC-MS method for detection of over 150 analytes in hops and a market survey of over 50 different hops pellets samples.
Abstract— Biofuel production from microalgae biomass appears as a promising long term alternative. Dunaliella tertiolecta is a microalgae with high tolerance to salinity, temperature, and light, making it relatively easy to grow. The aim of this study was to establish a pilot-scale culture to evaluate the biomass yield and bioethanol production. The cell culture of D. tertiolecta was started in 20 ml tubes and escalated to 20 L containers. The biomass yield was 0.153 g L-1 of dry basis (db) and its characterization showed protein (37% db) as major component followed by carbohydrates (35.6), lipids (13% db) and ash (6.5%). The carbohydrate fraction was composed of starch (27.1% db) and fiber (8.5 %) and its neutral sugar characterization yield glucose (91% molar). The main components of the lipid fraction were linolenic and palmitic acids. The biomass was subjected to an acid pre-treatment for the saccharification of complex carbohydrates, and the hydrolyzed biomass was fermented by Saccharomyces cerevisiae. It was possible to produce 0.615 ml g-1 of ethanol. In conclusion, D. tertiolecta has the potential for bioethanol production, making it a promising option for the biofuels future.
International Journal of Engineering and Science Invention (IJESI)inventionjournals
International Journal of Engineering and Science Invention (IJESI) is an international journal intended for professionals and researchers in all fields of computer science and electronics. IJESI publishes research articles and reviews within the whole field Engineering Science and Technology, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
Integration of sewage sludge digestion with advanced biofuel synthesiszhenhua82
The document describes integrating anaerobic digestion of sewage sludge with advanced biofuel production. Sewage sludge was treated with anaerobic digestion under two conditions: 1) low pH control and 2) chemical inhibition of methanogens. Both treatments resulted in accumulation of acetic acid. Acetic acid from digestion was then used as a carbon source for a fungus (Mortierella isabellina) and engineered Escherichia coli to produce fatty acids. The engineered E. coli strain had higher fatty acid yield and produced both medium and long chain fatty acids, while the fungus mainly produced long chain fatty acids. The study demonstrated a potential process to combine anaerobic digestion with microbial cultivation to simultaneously treat sewage
Degradation of an organophosphorus insecticide (chlorpyrifos) in simulated wa...Salah Hussein
Induced degradation of chlorpyrifos insecticide in simulated wastewater with advanced oxidation processes (AOPs), using ultraviolet irradiation (UV), ozonation and chemical oxidation using (sodium hypochlorite, calcium hypochlorite, monochloride-isocyanuric acid (MCICA), dichloroiso-cyanuric acid (DCICA), trichloroisocyanuric acid (TCICA) ) was studied. Chlorpyrifos and its degradation products were extracted using solid phase extraction (SPE) method, identified using GC-MS. Results showed that the degradation of chlorpyrifos in simulated wastewater followed the first order reaction, and its half life was 3.34, 5.64, 7.13 and 10.69h under ozonation, UV, 1.5%TCICA and 1.5%DCICA respectively when chlorpyrifos solutions treated for 12 h. The concentrations of chemical oxidative substances, active chlorine content and time of treatments had a significant effect on degradation rate of chlorpyrifos, which increased with increasing of each. The most enhancement of chlorpyrifos degradation was observed in treatment with ozonation, UV, TCICA and DCICA where the dissipations % of the parent compounds were 85.70, 57.71, 43.71 and 35.07 %, respectively. The intermediates products of chlorpyrifos degradation using chemical method were identified as O,O-Diethyl thiophosphate(DEP), 3,5,6-trichloro-2-pyridinol(TCP), 3,5,6-trichloro-2-methoxypyridine(TMP) and 2,3,5,6-tetrachloro-pyridine. UV leads to formation of O,O-Diethyl phosphate, TCP and Chlorpyrifos oxon. Ozonation leads to formation of O,O-Diethyl thiophosphate beside the UV degradation products.
54.Isolation and purification of cellulase from Aspergillus terreusAnnadurai B
This document describes the isolation and purification of cellulase enzymes from the fungus Aspergillus terreus. The intracellular cellulase was purified using ammonium sulfate precipitation, DEAE-cellulose chromatography, and gel filtration chromatography. This purification scheme achieved a 270-fold purification with a 22.11% yield. Tests including PAGE, SDS-PAGE, immunodiffusion, and isoelectric focusing confirmed the homogeneity of the purified enzyme. The purified cellulase showed optimal activity between pH 4-7 and temperatures of 40-50°C.
This document summarizes a study that investigated the physiological, biochemical, and genotoxic effects of wastewater on maize seedlings. The study exposed maize seeds and seedlings to different concentrations (0, 10, 50, and 100%) of wastewater collected from three sources: municipal wastewater, woolen mill wastewater, and polyvinylchloride wastewater. It found that the wastewater negatively impacted germination rates, biomass, seedling length, and photosynthetic pigments in a concentration-dependent manner. It also observed increases in oxidative stress markers and antioxidant enzyme activity in response to the wastewater. Various sources of wastewater were found to cause genotoxic effects
Similar to PRODUCTION AND OPTIMIZATION OF CHOLESTEROL OXIDASE FROM RHODOCOCCUS SPECIES (20)
Chronic Overworking: Cause Extremely Negative Impact on Health and Quality of...SUS GROUP OF INSTITUTIONS
Work is an action that organizes and provides meaning to the use of time in a society that
has programmed its rhythms as a function. It is important in structuring daily life and in
enabling a sense of continuity, provides capital, satisfaction that flourishing human life and
his family. What’s more, it is an antidote against boredom and emptiness. But it also
means we never really clock out while working and become too much workaholic. The
persistent overwork has extremely negative impacts on our health, happiness, and overall
quality of life. Nowadays working overtime has become the norm for most people. It is one
of those things everyone knows is bad for us, but no one really listens. Imbalance between
work and health or overwork not only bad for employees but also for employers. The long
working in the office or at home is bad for our health and our performance at work. A
person who expands more time in work may experience numerous health problems
including mental, physical and social problems. The Significant effects include stress, lack
of free time, poor work-life balance, relation hit and serious health risks lead to tiredness,
fatigue, obesity, lack of attentiveness, insomnia, depression, diabetes, high BP, Cerebrocardiovascular
problem, etc.
Anti-Oxidant and Antimicrobial Studies of Tinospora cordifolia (Guduchi/Giloy...SUS GROUP OF INSTITUTIONS
Plants produce a diverse range of bioactive molecules, making them a rich source of
different types of medicines and healing properties. The present study was aimed to
evaluate the anti-oxidant and antimicrobial properties of stem and root of T. cordifolia.
Total phenolic contents of different solvent extracts were determined and found that ethanol
extract had the highest phenolic content of 0.3213 mg g-1. Antioxidant assays were also
carried out by using different in vitro models such as total reducing power, hydrogen
peroxide scavenging activity assay and hydroxyl redical scavenging activity. The Ethanol
extract showed the highest total antioxidant activity. The H2O2 scavenging and hydroxyl
free radical scavenging activity was maximum 87.2 % and 91.0% found in case of ethanolic
steam extract respectively. The antimicrobial activity of ethanolic and methanolic extract of
root and stem of T. cordifolia were also evaluated against some pathogenic microorganisms
viz. E. coli, B. subtilis, A. niger and Candida sp. it was found that the various concentration
of extract viz. 50, 100, 150 and 200 mg ml-1 were tested. It was observed that the
increasing in concentration there was also increasing in antimicrobial activity reveled by
increase in size of zone of inhibition. The methanolic stem extract exhibits highest
antimicrobial activity against all four pathogens. The study shown that the extract of T.
cordifolia has a wide range of anti-oxidant as well as antimicrobial activity against bacterial
as well as fungal pathogens.
This study was conducted to establish bacterial contamination of cell phones and microbial contamination of
mobile phones and isolate the significant bacterial species associated with these cell phones in reference
to give necessary remedial measure. A total of 80 samples were collected to isolate microbial
population associated with cell phones. Sterile swabs were firmly rubbed on the surface of the handset, the
key buttons and on the screens of cell phones. The swabs were then inoculated into different media viz.
Nutrient agar, MacConkey agar, Mannitol Salt agar and Eosin Methelyne Blue agar. A total of 143
different bacterial isolates recovered from these sample and were classified as: Staphylococcus spp.
Corynebacterium spp., Streptococcus spp., Pseudomonas spp., Micrococcus spp., Proteus spp., Bacillus spp.,
and Enterobacter spp. at the ratio of 52, 17,14,7,4,3,2 and 1% respectively. The isolates were further
subjected for Antibiotic susceptibility profiling and have found that most of the recovered isolates were
challenging to Ampicillin, few isolates also shown intermediate results. Impimen, Norfloxacin and
Gentamycin were sensitive towards most isolates. Ciprofloxacin and Chloramphenicol showed variable
susceptibility to the different isolates. The study shown that all cell phones under investigation
were significantly contaminated by numerous bacterial species. It is an also indication that the majority of
them belongs to the normal flora of the human body as well as airborne and soil bacteria. Thus it can be said
that it is necessary to sterilise hands after contact with a cell phone since it is a potential source of disease
transmission.
Effect of Various Substrate and Process Parameters on the Production of Prodi...SUS GROUP OF INSTITUTIONS
In the present study it has been investigated that Serratia marcescens MTCC 4822 has
good potential for Prodigiosin production. Among the screened media components,
maltose was the best carbon source for the production for this strain. The fermentation
media supplemented with maltose (2%) and NaCl (0.5%) at pH 6.8-7.0 incubated at 28°C
gave maximum prodigiosin production (1390 unit/cell) with the biomass content of 3.45 g
L-1 after 96 hrs of incubation period. Prodigiosin, a red pigment, produced by bacterial
species Serratia marcescens, belongs to the family of tripyrrole was found to exhibit
antibacterial, antimycotic, immunomodulating, anti-tumor and anti-malarial properties. A
lot of attention is now paid to the biotechnological synthesis of the colours through the
microorganisms. Plant cell and tissue culture, microbial fermentation and gene
manipulation have been investigated with respect to the production of biopigments.
However, extensive safety testing of such products is required before they are given
clearance as safe food additives or other applications.
α-Galactosidase Producing Probiotics Bacteria and Their Health ImplicationsSUS GROUP OF INSTITUTIONS
Nowadays, people are aware that diet plays a major role in preventing diseases and promoting health.
Therefore there is an increasing trend for functional foods containing probiotic culture. “Probiotics are
defined as live microorganisms which when administered in adequate amounts confer a health benefit
on the host”. Some LAB positively influence human health mainly by improving the composition of
intestinal micro biota and for this reason, they are called probiotics. The increasing cost of health care,
the steady increase in life expectancy and the desire of the elderly for improved quality of life research
and development required in the area of probiotics. The concept of providing functional foods
including beneficial components rather than removing potentially harmful components. Soybeans
and other pulses contain oligosaccharides which may cause intestinal disturbances such as
flatulence. This study was undertaken to investigate α-galactosidase-producing probiotics bacteria.
The enzymes and cultures can be added to foods in order to enhance the digestibility of
carbohydrates in the gastrointestinal tract. However since many of these bacteria are reported for
probiotic properties that support and induced health benefits to the consumer. The study provides
data on the stability of α-galactosidase, which could potentially be added to food matrices
containing stachyose or raffinose such as beans, soya and other pulses and could be an alternative
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PRODUCTION AND OPTIMIZATION OF PECTINASE BY BACILLUS SP. ISOLATED FROM VEGETA...SUS GROUP OF INSTITUTIONS
Microbial enzymes have shown tremendous potential for different applications. Over the years due to their remarkable features enzymes have occupied the centre stage of all the biochemical and industrial processes. Pectinases are a group of enzymes responsible for the hydrolysis of pectic materials found in plants and are important industrial enzymes. In the present study, pectinase is produced from Bacillus sp. that was isolated from vegetable waste dump soil samples. A total of five isolates showed pectinase production and designated as PPB1 to PPB5. The screened isolates were used as a source of pectinase production using cassava waste as a substrate. Isolate PPB5 showed maximum enzyme activity of 0.641 IU/ml. Pectinase activity was optimized for various parameters like incubation time, temperature, pH, different carbon and nitrogen sources. Enzyme activity was observed maximum at 96 hr of incubation, 35°C temperature and at pH 6. The best carbon was found to be glucose. Among organic and inorganic nitrogen sources yeast extract and ammonium nitrate was founded to be better than other nitrogen sources. Among the five isolates, the isolate PPB5 showed maximum activity at all optimum conditions. This isolate is best producer and can be used in future for further pectinase production.
POTENTIAL BIOMEDICAL AND PHARMACEUTICAL APPLICATIONS OF MICROBIAL SURFACTANTSSUS GROUP OF INSTITUTIONS
Many microorganisms are able to produce a wide range of amphipathic
compounds, with both hydrophilic and hydrophobic moieties present
within the same molecule which allow them to exhibit surface
activities at interfaces and are generally called biosurfactants.
Biosurfactants are versatile, structurally diverse group of surface-active
substances produced by microorganisms and have variety of
applications in the sectors including bioremediation, food industry,
agriculture and pharmaceuticals. Interest in biosurfactant production
has markedly increased during the past decade, although large-scale
production has not been possible because of low production yields and
high total costs. At present, biosurfactants have gained importance in environmental
applications, while new applications in the pharmaceutical, biomedical, cosmetic and food
industry, with a high added value, are still developing. Recently, the potential applications of
biosurfactants in the biomedical field have increased. Their antibacterial, antifungal and
antiviral activities make them relevant molecules for applications in combating many
diseases and as therapeutic agents. In addition, their role as anti-adhesive agents against
several pathogens indicates their utility as suitable anti-adhesive coating agents for medical
insertional materials leading to a reduction in a large number of hospital infections without
the use of synthetic drugs and chemicals. This article emphasizes the medicinal and
therapeutic perspective of biosurfactants. With these specialized and cost-effective
applications, biosurfactants can be considered as an interesting option for the near future.
A REVIEW ON APPLICATIONS OF BIOSURFACTANTS PRODUCED FROM UNCONVENTIONAL INEXP...SUS GROUP OF INSTITUTIONS
Biosurfactants can serve as green alternative in different areas due to
their ecological acceptance as they are biodegradable and nontoxic.
Nowadays biosurfactants are predominantly used in pharmaceutical,
oil industry, and for the bioremediation of pollutants. Apart from these,
biosurfactants also show potential applications in many sectors of food
industry and agriculture. Allied with emulsion forming and breaking,
antiadhesive, functional ingredient, are some properties that can be
exploited in agro-food biotechnology. Potential role of biosurfactants
in food and agricultural sectors as well as present concern of lowering
the production cost of biosurfactants by using the unconventional
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Phytochemical, Antioxidant and Antibacterial Studies on Bambusa arundinacea a...SUS GROUP OF INSTITUTIONS
This study was formulated to check the phytochemical, antioxidant, antibacterial potential of
Bambusa arundinacea (Bamboo) and Mangifera indica (Mango) trees. Aqueous, ethanolic and
methanolic extracts were prepared from leaves of former and stem bark of later. The phytochemical
screening of the extracts showed the presence of various bioactive compounds such as
carbohydrates, flavonoids, saponins and proteins in B. arundinacea, alkaloids, flavonoids, tannins,
saponins, steroids and cardiac glycosides in M. indica. Total phenolic concentration and
percentage of free radical scavenging activity was more in ethanolic extracts of B. arundinacea and
M. indica followed by methanolic extracts and aqueous extracts. Highest percentage of ferric
reducing antioxidant power was found in ethanolic extracts and lowest in aqueous extracts indicates
that ethanolic extracts has more antioxidant potential than the other two extracts. Ethanolic extracts
of both plants had higher inhibition on the tested Gram positive (B. subtilis & S. aureus) as well as
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methanolic extracts of both the tested plants were more effective than aqueous extracts due to better
extraction power of organic solvents. Overall study indicates that B. arundinacea and M. indica are
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development of novel drugs and may represent new source of antimicrobials with stable, biologically
active components that can establish a scientific base for further use in modern medicines.
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PRODUCTION AND OPTIMIZATION OF CHOLESTEROL OXIDASE FROM RHODOCOCCUS SPECIES
1. PRODUCTION AND OPTIMIZATION OF CHOLESTEROL OXIDASE FROM RHODOCOCCUS SPECIES
Original Article
SUKHVIR KAUR*, HARJOT PAL KAUR, BHAIRAV PRASAD, METAN PRASHER
Department of Biotechnology, SUS College of Research and Technology, Tangori (Mohali), Punjab, India
Email: sukhvirkaur29@gmail.com
Received: 18 Apr 2015 Revised and Accepted: 21 May 2015
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Objective: Optimization of conditions for cholesterol oxidase production by the microorganism isolated from urban compost and dairy soil samples.
Methods: Isolates were obtained on the basis of their capability of growing on isolation medium A and B and their cholesterol oxidase (CHO) production
was estimated. CHO production was optimized by the optimization of temperature, pH, carbon sources, and organic and inorganic nitrogen sources.
Results: 3 isolates out of 22 were found to secrete extracellular CHO as detected by cholesterol oxidase indicator plate A and were designated as
cholesterol oxidase producing isolate 1, 2 and 3 (COP 1, COP 2 and COP 3). Results showed that the strain COP 2 belonging to the genus Rhodococcus
sp. based on morphological, cultural and biochemical characteristics recorded highest cholesterol oxidase activity. Optimum temperature and pH
for CHO activity were found to be 35 °C and 7.5 respectively. Steroidal substrate cholesterol produced a significant increase in CHO level (0.502
IU/ml). Organic and inorganic nitrogen sources were supplemented in combinations leads to increase in CHO production as compared to individual
components. (NH4)2HPO4
Conclusion: The isolate COP 2 produced significant levels of cholesterol oxidase extracellularly in optimized medium as compared to cell bound
CHO, and can be easily produced on an industrial scale.
and yeast extract supported the highest enzyme production (0.574 IU/ml).
Keywords: Cholesterol oxidase, Rhodococcus sp, Biochemical, Optimization.
INTRODUCTION
Cholesterol oxidase (COX, EC 1.1.3.6) a monomeric bi-functional
FAD-containing (flavo enzyme) enzyme belongs to the family of
oxidoreductases, specifically those acting on the CH-OH group of
donors with oxygen as acceptor. Cholesterol oxidase (CHO) enzyme
catalyzes the oxidation of cholesterol and converts 5-cholesten-3-ol
into 4-cholesten-3-one [1]. The systematic name of this enzyme class
is cholesterol: oxygen oxidoreductases and other common names in
use are 3β-hydroxy steroid oxidoreductases, and 3β-hydroxy
steroid: oxygen oxidoreductases. CHO was first isolated and
characterized from Rhodococcus erythropolis and has many
applications in agriculture, medicinal industry and pharmaceutical
sectors. For instance, it can be used for the production of diagnostic
kits to detect blood cholesterol, biological insecticide and precursors
for steroid hormones [2, 3]. Cholesterol oxidase enzyme is simple,
specific, and highly sensitive; its use has become widespread in the
determination of serum cholesterol that has direct implications in
atherosclerosis, coronary heart disease and other lipid disorders,
and for determining the risk of heart attack and thrombosis. This
enzyme shows potent insecticidal activity has been used to track cell
cholesterol and has also been found to be a potent parricide [4, 5]. It
has also been developed as a pest control in the agricultural industry
especially in transgenic crops [4]. Biochemical and structural,
studies of enzyme revealed the involvement of an enzyme in
interaction with lipid bilayer [6, 7]. With only the exception of glucose
oxidase, CHO is the most widely used enzyme in clinical laboratories.
Many bacteria can produce this enzyme including members of the
genera Arthrobacter, Brevibacterium, Pseudomonas, Nocardia,
Rhodococcus, Streptomyces, Corynebacterium and Shizophylum [8]. This
enzyme can be produced by a bacterium in three forms: intracellular,
extracellular and membrane bound. Due to the wide spectrum
applications of CHO, screening and isolation of bacterial strains
producing extracellular form of CHO are of major importance [9].
Because of the commercial value of cholesterol oxidase there is wider
interest in producing this enzyme from microbial cells. Thus, there is
need to isolate newer bacterial cultures to produce cholesterol oxidase
optimally under physiological conditions and to study the important
biochemical properties of this enzyme.
Therefore, keeping in mind the importance of cholesterol oxidase
enzyme the present study was planned to isolate CHO enzyme
producing bacteria from urban compost and dairy soil sample, to
determine the type (extracellular or cell bound) of CHO enzyme
produced by the isolates and to optimize cholesterol oxidase
production by optimizing the cultural conditions.
MATERIALS AND METHODS
Collection of soil samples
26 different urban compost and dairy soil samples were collected
from Himachal Pradesh, Jammu & Kashmir and Punjab in a sterile
polythene bags and brought at the laboratory for further studies.
Isolation of bacteria
For the isolation of microorganisms, 1 g of soil samples from
different places were suspended in 100 ml of distilled water and was
shook vigorously for 30 mins by placing it in an incubator shaker. A
volume of 100 µl of supernatant was inoculated onto the medium
“A” (contained (g/l): agar, 3.0%; K2HPO4, 0.25; NH4NO3, 17; MgSO4.
H2O 0.25 % ; FeSO4
Screening of CHO producing bacteria
, 0.001; NaCl, 0.005; cholesterol, 0.1 % and
Tween 80, 0.5 ml) at pH 7. The inoculated plates were placed in the
incubator for 7-10 days at 30 ˚C. After incubation, plates were
observed for colony characteristics. Larger fast growing colonies
were sub cultured in secondary medium containing cholesterol as
the only source of carbon as well as yeast extract [9-10].
CHO producing colonies were selected on cholesterol oxidase
indicator plates. These plates were prepared by adding 1.0 g
cholesterol, 1.0 g triton X-100, 0.1 g o-dianisidine and 1000 U of
peroxidase to one l of agar medium. Bacterial colonies were cultured
on these plates and incubated at 30 ˚C. Cholesterol penetrates into
bacterial cells where it can be converted into hydrogen peroxide by
cholesterol oxidase. Reagents that exist in the medium react with
hydrogen peroxide (H2O2
Identification of isolates
) to form azo compound which turns the
color of medium into intense brown color [11-13].
CHO producing isolates were identified by their morphological,
cultural and biochemical characteristics by standard methods using
Bergey’s manual of systematic bacteriology [14].
International Journal of Pharmacy and Pharmaceutical Sciences
ISSN- 0975-1491 Vol 7, Issue 7, 2015
Innovare
Academic Sciences
2. Kaur et al.
Int J Pharm Pharm Sci, Vol 7, Issue 7, 265-268
266
Assessment of enzyme activity
CHO activity of isolated bacteria was detected by hydrogen
peroxide generated during cholesterol oxidation process and was
measured [15]. In this reaction, hydrogen peroxide was coupled
with 4-aminoantipyrine and phenol by peroxides to produce
quinoneimine dye with maximum absorption in 500 nm. Dissolved
cholesterol in non-ionic detergent triton X-100 was used as the
substrate of the reaction. The reaction mixture was consisted of 3
µmol of cholesterol in 1.0 ml of 1% triton X-100, 300 µmol of
phosphate buffer (disodium phosphate-monopotassium
phosphate), pH 7.0, 0.1 ml of enzyme solution, 1.2 µmol of 4-
aminoantipyrine, 21 µmol of phenol and 20 U of horseradish
peroxidase (HRP) in a final volume of 3 ml. Reaction was
performed at 37 ˚C for 10 min with shaking during incubation
period. This reaction was terminated by heating at 100 ˚C for 3
min. One unit of enzyme was defined as the amount of enzyme that
forms 1 μmol of H2O2 per minute at 37 ˚C.
To determine the cell bound and extracellular CHO activity, bacterial
cultures were centrifuged in 10,000 x g for 5 min and supernatant
was used as an extracellular enzyme source. In order to find the cell
bound CHO, cellular pellet was washed twice with 0.1 M phosphate
buffer (pH 7.2). After resuspending cellular pellet in the same buffer
containing 0.1% of triton X-100, the supernatant was incubated at
30 ˚C for 30 mins, centrifuged in 10,000 x g and used for detection of
cell bound CHO [9].
Effect of pH on CHO activity
To evaluate the optimum pH for the enzyme activity, the enzyme
solution was incubated from pH 4.5 to 9 for 24 h and enzyme
activity were measured under standard conditions. Acetate buffer
was used for pH 4.5 to 5.5, phosphate buffer for pH 6 to 7 and Tris-
HCl for pH 8 to 9 [8, 15].
Effect of temperature on CHO activity
In order to determine the effect of temperature on enzyme activity
Sasaki's method was performed on an enzyme solution under standard
conditions except for reaction temperature. The enzyme solution was
incubated at 20, 25, 30, 35, 40, 45 and 50 ˚C for 24 h [8, 15].
Effect of carbon sources
Carbon sources (0.2% w/v) glucose, steroidal compound (cholesterol)
and non-steroidal hydrophobic substrate (n-hexanoate) were
supplemented as individual components to the production media to
check the effect of these sources on enzyme production [16].
Effect of nitrogen sources
Combination of inorganic (0.2% w/v) and organic (0.5% w/v)
nitrogen sources as well as an individual organic component (yeast
extract) and inorganic component (NH4)2HPO4
RESULTS AND DISCUSSION
(Diammonium
hydrogen phosphate) were checked for CHO production [16].
22 bacterial isolates were obtained from 26 urban compost and
dairy site soil samples collected from various sites based on their
capability to grow on an isolation medium A containing cholesterol
as the sole carbon source. Among them, only 3 isolates were found
to cholestrol oxidase producing (COP) and designated as COP 1, COP
2 and COP 3. Then, these strains were examined for CHO production
and results showed that the isolated strain COP 2 had the highest
ability for CHO production (1.162 U/ml) in comparison with the
other isolates in extracellular form as compared to cell bound
(Table-1). Therefore, strain COP 2 was chosen for further studies.
The cells of COP 2 were gram positive, irregular rods but eventually
presented as coccoid forms as growth continued changed and
results of biochemical characteristics are shown in table 2. From
these observations, it is clear that isolate COP 2 belongs to
Rhodococcus species. In previous studies also a high amount of CHO
enzyme produced by Rhodococcus is extracellular form and only a
low amount of CHO is membrane bound or intracellular type [17].
Table 1: Extracellular and cell bound CHO activity of isolates
S. No. Isolate Extracellular activity (U/ml) Cell bound activity (U/ml)
1. COP 1 0.786±0.08 0.452±0.05
2. COP 2 1.162±0.09 0.964±0.05
3. COP 3 0.926±0.04 0.654±0.03
Values are means±SD of three replicates (n=3)
Table 2: Biochemical properties of isolate COP 2
Test Result Test Result
Gram reaction Positive H2 VariableS production test
Catalase Positive Lactose fermentation Negative
Haemolysis Negative Sucrose fermentation Negative
Oxidase Positive Maltose fermentation Negative
Blood agar culture Mucoid colonies Xylose fermentation Negative
Indole production Negative Glucose fermentation Negative
Methyl Red test Negative Mannitol fermentation Negative
Vogus Proskaur test Negative Motility Negative
Nitrate reduction test Negative Endospore Absent
Urease production test Negative Gelatin hydrolysis Negative
Effect of temperature
Enzyme activity was measured under standard conditions except for
reaction temperature. Optimum CHO activity was seen at 35 ˚C from
Rhodococcus sp. (fig. 1). Results of the present study were
compatible with an earlier study conducted by Kumari and Kanwar
[18]. Temperature 35 ˚C was found to be optimum for Bacillus
licheniformis [19] and the optimum activity and stability of CHO
from Streptomyces fradiae and Brevibacterium sp. were reported at
50 ˚C and 53 °C for 30 min. respectively [9, 20]. Optimum
temperature for maximum enzyme activity from Rhodoccocus equi
and from Corynebacterium cholesterolicum, was 47 ˚C and 40 ˚C
respectively [21] and the optimum enzyme production in the basal
medium was 30 °C [16].
Effect of pH
The optimum pH for production of CHO enzyme was recorded to be
pH 7.5 as maximum CHO production (0.498 U/ml) was recorded at
this pH (fig. 2). Results of the present study were compatible with
3. Kaur et al.
Int J Pharm Pharm Sci, Vol 7, Issue 7, 265-268
267
previous studies in which pH 7 was noted optimum for enzyme
production [19, 20, 16]. The optimum pH for CHO production in
bound form without centrifugation was 6 and after centrifugation
were 6 and 8 respectively [22].
Fig. 1: Effect of incubation temperature on enzyme production
Fig. 2: Effect of pH on enzyme production
Effect of carbon sources
CHO was produced with all the carbon sources tested; however,
steroidal substrate cholesterol (0.502U/ml) only showed a
significant increase in CHO levels (fig. 3). Results of the present
investigation were similar to the results of Ahmad and Goswami [16]
who also observed steroidal substrate to increase levels of CHO
production. Rhodococcus erythropolis, under appropriate conditions,
can show a significant CHO activity when grown in a mineral
medium containing cholesterol as a sole carbon source [23].
Fig. 3: Effect of carbon sources on enzyme production
Effect of nitrogen sources
Different combinations of inorganic and organic nitrogen sources
were supplemented to CHO production and it was observed that
yeast extract with the combination of (NH4)2HPO4 give maximum
CHO production (0.574I U/ml). Organic (yeast extract) and
inorganic (DAPH) nitrogen sources when supplemented as
individual components to the production media leads to decrease in
enzyme production (0.286 and 0.25 U/ml respectively) (fig. 4).
Earlier study also reported that the combination of (NH4)2HPO4 and
yeast extract leads to the highest enzyme production (0.353 U/ml)
as compared to individual organic and inorganic components [16].
Fig. 4: Effect of inorganic and organic nitrogen sources on
enzyme production
A. A (ammonium acetate), A. C(ammonium chloride), A. N
(ammonium nitrate)
CONCLUSION
, A. S (ammonium sulphate), DAHP (di
ammonium hydrogen phosphate), Y. E(yeast extract), B. E (beef
extract), M. E (malt extract)
Cholesterol oxidase is an enzyme of great commercial value widely
employed by laboratories routinely devoted to the determination of
cholesterol in food, serum and other clinical samples. Rhodococcus
species which are capable of producing cholesterol oxidase have been
isolated from urban compost and dairy soil samples. Bacterial isolate
COP 2 showed maximum enzyme production and production was
increased by optimization of cultural conditions viz., temperature, pH,
carbon and nitrogen sources. Our work led to the conclusion that
isolated Rhodococcus sp. might be considered as potentially interesting
source of cholesterol oxidase for commercial purpose.
CONFLICT OF INTERESTS
Declared None
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