This document describes research on the production and characterization of carboxymethyl cellulase (CMCase) from Paenibacillus polymyxa using mango peel as a substrate. Key findings include:
- P. polymyxa exhibited maximum CMCase production when grown in a medium containing 7% mango peel with 1.5% ammonium sulfate at 37°C and pH 5.5.
- Purification by affinity column chromatography achieved a 28-fold purification with a 1.99% recovery yield.
- SDS-PAGE analysis showed bands at 26.5 and 34 kDa, suggesting a heteromeric multienzyme complex. Native PAGE showed a single band of 72 kDa.
IJERA (International journal of Engineering Research and Applications) is International online, ... peer reviewed journal. For more detail or submit your article, please visit www.ijera.com
Microbial Processing of Agricultural Wastes to produce Pectinase Enzyme(s) an...Meesha Singh
Microbial Processing of Agricultural Wastes to produce Pectinase Enzyme(s) and Evaluation of their Industrial Applications
This document summarizes a study on the production of pectinase enzymes from agricultural wastes using microbial processing. Pectinases are important industrial enzymes with applications in juice extraction and clarification. The study aims to optimize pectinase production from bacterial sources using agricultural waste, characterize the enzymes, and evaluate their applications. Key steps include screening bacteria, optimizing fermentation parameters, purifying and characterizing the enzymes, and testing applications in juice clarification and fruit peeling. A hierarchical model is also developed to analyze barriers to industrial pectinase production.
Isolation and characterization of polyphenol oxidase from phyllanthus emblica...IJSIT Editor
This study aimed to isolate and characterize polyphenol oxidase (PPO) from Phyllanthus emblica (Indian gooseberry). PPO was purified from gooseberry extracts using ammonium sulfate precipitation and dialysis. SDS-PAGE analysis showed the purified PPO had an apparent molecular weight of 100 kD. Optimum conditions for PPO activity were determined to be pH 7 and temperatures between 30-40°C. PPO showed highest activity with catechol and was inhibited by EDTA, indicating it is a copper-containing enzyme. Characterization of gooseberry PPO provides insights into browning of this fruit.
IOSRPHR(www.iosrphr.org) IOSR Journal of Pharmacyiosrphr_editor
Vigna radiata β-amylase was immobilized onto woven Bombyx mori silk fabric activated with sodium nitrate and chlorination. The immobilization resulted in 90% retention of enzyme activity. The immobilized enzyme had improved thermal stability up to 72°C compared to 40°C for free enzyme. It also had good storage stability, maintaining 60% activity over 3-4 months. The optimum conditions for the immobilized enzyme were determined, including incubation time, pH, temperature, substrate concentration, and CaCl2 concentration.
Assessing the Suitability of using Plant Latex as Immobilization Support for ...ijsrd.com
Horseradish peroxidase was immobilized onto latex from three different plants viz. Calotropis procera, Euphorbia royleana and Alstonia scholaris with 0.51 ± 0.01, 0.37 ± 0.01, 0.46 ± 0.01 mg/cm2 conjugation yield and 62.07 ± 0.85, 66.1 ± 0.85, 71.24 ± 0.80 % retention of specific activity respectively. The support, before and after addition of peroxidase was characterized using scanning electron microscopy (SEM) and Fourier transmission infra-red spectroscopy (FTIR). Optimum pH, optimum temperature and changes in kinetic parameters (Ea, Km and Vmax) for immobilized peroxidases were studied and found to differ from that of free peroxidase. Alstonia scholaris latex was most effective in stabilizing the structure of peroxidase during storage at 4°C, whereas thermal stability and reusability of peroxidase was better on Calotropis procera latex. Analytical use of Calotropis procera latex bound peroxidase for determination of phenolic content of fruit juices has also been demonstrated.
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.
This document summarizes research that optimized production of exopolysaccharide (EPS) by Pseudomonas fluorescens using rice bran as a carbon source. Plackett-Burman statistical design was used to screen significant medium components, identifying rice bran, peptone, NaCl, and MnCl2 as most effective. Central composite design then optimized their concentrations for maximum EPS yield of 4.62 g/L after 48 hours of fermentation. Kinetic modeling using logistic growth and Luedeking-Piret equations accurately simulated microbial growth, substrate utilization, and EPS formation. The extracted polysaccharide shows potential as a biodegradable, non-toxic alternative to synthetic polymers with antioxidant properties for industrial and pharmaceutical
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
IJERA (International journal of Engineering Research and Applications) is International online, ... peer reviewed journal. For more detail or submit your article, please visit www.ijera.com
Microbial Processing of Agricultural Wastes to produce Pectinase Enzyme(s) an...Meesha Singh
Microbial Processing of Agricultural Wastes to produce Pectinase Enzyme(s) and Evaluation of their Industrial Applications
This document summarizes a study on the production of pectinase enzymes from agricultural wastes using microbial processing. Pectinases are important industrial enzymes with applications in juice extraction and clarification. The study aims to optimize pectinase production from bacterial sources using agricultural waste, characterize the enzymes, and evaluate their applications. Key steps include screening bacteria, optimizing fermentation parameters, purifying and characterizing the enzymes, and testing applications in juice clarification and fruit peeling. A hierarchical model is also developed to analyze barriers to industrial pectinase production.
Isolation and characterization of polyphenol oxidase from phyllanthus emblica...IJSIT Editor
This study aimed to isolate and characterize polyphenol oxidase (PPO) from Phyllanthus emblica (Indian gooseberry). PPO was purified from gooseberry extracts using ammonium sulfate precipitation and dialysis. SDS-PAGE analysis showed the purified PPO had an apparent molecular weight of 100 kD. Optimum conditions for PPO activity were determined to be pH 7 and temperatures between 30-40°C. PPO showed highest activity with catechol and was inhibited by EDTA, indicating it is a copper-containing enzyme. Characterization of gooseberry PPO provides insights into browning of this fruit.
IOSRPHR(www.iosrphr.org) IOSR Journal of Pharmacyiosrphr_editor
Vigna radiata β-amylase was immobilized onto woven Bombyx mori silk fabric activated with sodium nitrate and chlorination. The immobilization resulted in 90% retention of enzyme activity. The immobilized enzyme had improved thermal stability up to 72°C compared to 40°C for free enzyme. It also had good storage stability, maintaining 60% activity over 3-4 months. The optimum conditions for the immobilized enzyme were determined, including incubation time, pH, temperature, substrate concentration, and CaCl2 concentration.
Assessing the Suitability of using Plant Latex as Immobilization Support for ...ijsrd.com
Horseradish peroxidase was immobilized onto latex from three different plants viz. Calotropis procera, Euphorbia royleana and Alstonia scholaris with 0.51 ± 0.01, 0.37 ± 0.01, 0.46 ± 0.01 mg/cm2 conjugation yield and 62.07 ± 0.85, 66.1 ± 0.85, 71.24 ± 0.80 % retention of specific activity respectively. The support, before and after addition of peroxidase was characterized using scanning electron microscopy (SEM) and Fourier transmission infra-red spectroscopy (FTIR). Optimum pH, optimum temperature and changes in kinetic parameters (Ea, Km and Vmax) for immobilized peroxidases were studied and found to differ from that of free peroxidase. Alstonia scholaris latex was most effective in stabilizing the structure of peroxidase during storage at 4°C, whereas thermal stability and reusability of peroxidase was better on Calotropis procera latex. Analytical use of Calotropis procera latex bound peroxidase for determination of phenolic content of fruit juices has also been demonstrated.
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.
This document summarizes research that optimized production of exopolysaccharide (EPS) by Pseudomonas fluorescens using rice bran as a carbon source. Plackett-Burman statistical design was used to screen significant medium components, identifying rice bran, peptone, NaCl, and MnCl2 as most effective. Central composite design then optimized their concentrations for maximum EPS yield of 4.62 g/L after 48 hours of fermentation. Kinetic modeling using logistic growth and Luedeking-Piret equations accurately simulated microbial growth, substrate utilization, and EPS formation. The extracted polysaccharide shows potential as a biodegradable, non-toxic alternative to synthetic polymers with antioxidant properties for industrial and pharmaceutical
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
This document discusses a study that characterized Ghanaian cocoa bean fermentation using spectroscopic and chromatographic methods. Researchers used colorimetry, fluorescence spectroscopy, near-infrared spectroscopy, and gas chromatography-mass spectrometry to examine cocoa beans sampled at different stages of fermentation. The degree of fermentation could be described well by the spectroscopic methods. Certain aroma compounds like 2-phenylethyl acetate increased during fermentation while others like diacetyl decreased. The study demonstrates the potential of using spectroscopy to objectively determine cocoa bean quality and degree of fermentation.
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
This document reports on a comparative study of the chemical compositions and feeding values of leaves from Moringa stenopetala and Moringa oleifera trees. The study found that M. oleifera leaves contained higher levels of crude protein, fat, essential amino acids, and gross energy than M. stenopetala leaves. However, M. stenopetala leaves had higher fiber fractions like crude fiber, acid detergent fiber and cellulose. In vitro gas production analysis showed M. stenopetala leaves produced more gas and had higher metabolizable energy, organic matter digestibility and short chain fatty acids than M. oleifera leaves. Both Moringa species showed potential as protein supplements for ruminants and non
Radiation Processing And Functional Properties Of Soybean (Glycine Max)Amit K. Das
This article discusses the effects of radiation processing on soybean and its functional properties. Radiation processing at doses of 10, 20, and 30 kGy reduced the cooking time of soybean and increased the oil absorption capacity of soy flour without affecting proximate composition. Irradiation also improved the functional properties of soybean protein isolate, including solubility, emulsification activity, and foam stability. These changes enable uses of soybean in products like soy milk, tofu, and tofu-fortified patties. The value addition effects of radiation processing are beneficial for better utilization of soybean.
This document provides biographical information on Madhav P. Yadav including his education, work experience, professional activities, awards, and publications. It can be summarized as follows:
Yadav has a Ph.D. in Biochemistry from Southern Illinois University and has worked as a research scientist at various academic and government institutions. He has over 70 publications and has received awards for his research focusing on carbohydrate chemistry and the development of value-added products from agricultural residues.
This document discusses medium optimization and antioxidant activity of exopolysaccharide produced by Bacillus subtilis. Bacillus subtilis was isolated from soil and used to produce exopolysaccharide. Statistical methods like Plackett-Burman design and response surface methodology using central composite design were used to optimize the culture medium. The optimized medium contained specific concentrations of cane molasses, yeast extract, NaCl, and CaCl2, which produced 4.92 g/L of exopolysaccharide. The exopolysaccharide showed antioxidant activity, with a 61.19% reduction of DPPH free radicals, greater than vitamin C. The biopolymer could have applications in industries and pharmaceuticals.
This study aimed to isolate thermostable cellulases from woodland soil samples using metagenomic approaches. Genomic DNA was extracted from soil samples and used to amplify fragments of endo-β-1,4-glucanases. Twenty-five different cellulase sequences were identified, with nine cloned and tested for cellulolytic activity at 60°C. One sequence, CelMS6, displayed the highest activity at 60°C, making it a good candidate for applications requiring cellulose utilization at high temperatures.
Enhanced endoglucanase production by Bacillus aerius on mixed lignocellulosic...Mushafau Adebayo Oke
Oke, M. A., Annuar, M. S. M., and Simarani, K. (2016). "Enhanced endoglucanase production by Bacillus aerius on mixed lignocellulosic substrates." BioResources, 11(3), 5854-5869.
Isolation and purification of peroxidase from soyabeanPooja Walke
Peroxidase (EC. 1.11.1.7), an oxidoreductase, has iron porphyrin ring generally and catalyzes a redox reaction between H202 as an electron acceptor and many kinds of substrates by means of oxygen liberation from HzOz (Brill, 1996).
Production of Amphiphilic Surfactant Molecule From Saccharomyces Cerevisiae M...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.
Lipase catalyzed process for biodiesel production protein engineering and lip...Melike Şeyma Kadayıfçı
This review document discusses the use of lipases in biodiesel production. It covers lipase engineering approaches like directed evolution and rational design to improve properties such as thermostability and activity. It also discusses lipase production methods like host strain selection and metabolic engineering. The goal is to develop high-performance lipases through these techniques to enable large-scale commercialization of lipase-catalyzed biodiesel production as an alternative to fossil fuels.
This document discusses optimizing the production of exopolysaccharide (EPS) by Bacillus subtilis using various carbon and nitrogen sources. Sucrose at 2% concentration produced the highest yield of EPS. Cane molasses and rice bran were also tested as carbon sources, with cane molasses at 2% giving the highest yield. Different solvents were tested for their ability to precipitate EPS, with ethanol, diethyl ether, and methanol being effective. Fourier transform infrared spectroscopy analysis revealed the extracted polymer was composed of sucrose units. In conclusion, agro-wastes like cane molasses could be used as alternative carbon sources for the economical production of EPS.
This document summarizes several research studies:
1. One study investigated the metabolic flexibility of Pseudomonas taiwanensis VLB120 by knocking out NADH dehydrogenases and performing multi-omics analysis. It revealed the bacterium's high robustness and proposed a metabolic response to type-2 NADH dehydrogenase deficiency.
2. Another characterized the BorB thioesterase from Streptomyces parvulus, determining its structure provides a substrate binding site and specificity.
3. A third engineered Pseudomonas putida to produce bisdemethoxycurcumin by leveraging its native lignin catabolism pathways.
4. A fourth studied Aspergillus niger's
Molecular Characterization of Isolated Methyl Parathion Degrading Bacteria an...IJSRD
The burning problem in present era is pesticide residue in fruits and foods. present study focus on degradation of pesticide in contest to that total 45 methyl parathion(MP) degrading bacteria were isolated from sludge of MP producing industrial waste. According to screening 21 highly efficient pesticide degraders were selected at 500ppm concentration of standard methyl parathion as a sole carbon source. All isolates were characterized by RAPD and based on the RAPD result all 21 bacterial isolates were grouped in to 2 main clusters with 58% similarity. Characterization of MP degrading gene was carried out by using specific primer. Out of 21 isolates, 14 isolates were containing mpd gene and 4 isolates were containing opd gene while remaining 3 isolates which did not show amplification with opd/ mpd primer so it may contain other unreported gene responsible for methyl parathion degradation. Isolates were identified based on 16s rRNA sequence and one highly efficient bacterium selected for genome characterization.
1. The study characterized an apple polyphenol extract standardized to 5% phloridzin and evaluated its effects on glucose transport.
2. In vitro, the extract showed a dose-dependent inhibition of up to 60% on glucose uptake by cells, comparable to known inhibitors.
3. In mice, the extract had a lower acute toxicity than green tea extract when administered orally or intraperitoneally.
4. The results suggest the extract may help regulate blood glucose levels and have potential benefits for managing diabetes and body weight control through inhibition of intestinal glucose transport. Further human studies are needed to confirm these effects.
Recent advances in technoeconomic analysis (TEA) were reviewed:
- TEA is useful for process design, cost estimation, and identifying bottlenecks early in research.
- Studies now enable faster iteration, robust uncertainty analysis, and open-source platforms.
- Trends include more expansive system boundaries and potential integration with high-throughput experiments.
Typha grass is an invasive wetland plant with high annual productivity that can be used for biorefinery. It contains high levels of sugars and low lignin, making it suitable for conversion processes. The objective is to explore Typha capensis's potential as a biorefinery feedstock. A two-step pretreatment was performed and yielded good enzymatic digestibility of sugars. Characterization of Typha lignin showed a higher S/G ratio, indicating easier delignification. Typha deserves consideration as a promising biomass feedstock for tropical biorefining due to its high productivity, simple pretreatment requirements, and good enzymatic hydrolysability.
This document describes a study that optimized media components for xylanase production by Aspergillus fumigatus using solid state fermentation. Key findings:
1. Plackett-Burman design identified three significant nutrients (NaNO3, (NH4)2SO4, KH2PO4) influencing xylanase production.
2. Central composite design and response surface methodology were used to further analyze the effects of these nutrients and establish a regression model.
3. Under optimized conditions found by response surface analysis (NaNO3 = 0.29066 g/gds, (NH4)2SO4 = 0.14448 g/gds, KH2PO4 0
Hydrolytic activity of amylase produced in solid state fermentation by a lo...Samuel Kwatia
This document summarizes research on the production and characterization of amylase enzyme from a strain of Aspergillus niger using solid-state fermentation. Key findings include:
- Amylase was produced using optimized fermentation conditions and showed highest activity at pH 4-8 and temperatures of 30-60°C.
- Partial purification using ammonium sulfate increased the specific activity and concentration of amylase.
- The purified enzyme was able to hydrolyze starches from various sources like maize, yam and cassava, producing reducing sugars.
- Different starch sources showed varying susceptibility to hydrolysis, with maize starch being most easily broken down.
A comparative study of biopolymers and alum in the separation and recovery of...Soumyadeep Mukherjee
This study compares the effectiveness of various biopolymers and alum in recovering pulp fibers from paper mill wastewater through flocculation. Guar gum was found to be the most effective biopolymer, recovering 3.86 mg/L of fibers. Alum displayed the highest particle removal rate. Guar gum, alum, xanthan gum, and locust bean gum removed 94.68%, 97.46%, 92.39%, and 92.46% of turbidity, respectively, at a settling velocity of 0.5 cm/min. Response surface methodology was used to optimize conditions like pH, dose, and mixing speed to obtain the lowest sludge volume index for guar gum. The results indicate similar performance
Pomelo peels as alternative substrate for extracellular pectinase production ...Haritharan Weloosamy
This document summarizes a study that investigated using pomelo peels as an alternative substrate for producing pectinase enzymes through solid state fermentation with the fungus Aspergillus niger HFM-8. Initial experiments found the highest pectinase production and fungal growth on day 8 of cultivation. Various culture parameters were then optimized, finding that a substrate particle size of 0.75 mm produced the highest pectinase levels and fungal growth. After improving the fermentation parameters, pectinase production increased by 48.82% compared to initial levels, demonstrating that pomelo peels are a suitable substrate for pectinase production.
This document discusses a study that characterized Ghanaian cocoa bean fermentation using spectroscopic and chromatographic methods. Researchers used colorimetry, fluorescence spectroscopy, near-infrared spectroscopy, and gas chromatography-mass spectrometry to examine cocoa beans sampled at different stages of fermentation. The degree of fermentation could be described well by the spectroscopic methods. Certain aroma compounds like 2-phenylethyl acetate increased during fermentation while others like diacetyl decreased. The study demonstrates the potential of using spectroscopy to objectively determine cocoa bean quality and degree of fermentation.
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
This document reports on a comparative study of the chemical compositions and feeding values of leaves from Moringa stenopetala and Moringa oleifera trees. The study found that M. oleifera leaves contained higher levels of crude protein, fat, essential amino acids, and gross energy than M. stenopetala leaves. However, M. stenopetala leaves had higher fiber fractions like crude fiber, acid detergent fiber and cellulose. In vitro gas production analysis showed M. stenopetala leaves produced more gas and had higher metabolizable energy, organic matter digestibility and short chain fatty acids than M. oleifera leaves. Both Moringa species showed potential as protein supplements for ruminants and non
Radiation Processing And Functional Properties Of Soybean (Glycine Max)Amit K. Das
This article discusses the effects of radiation processing on soybean and its functional properties. Radiation processing at doses of 10, 20, and 30 kGy reduced the cooking time of soybean and increased the oil absorption capacity of soy flour without affecting proximate composition. Irradiation also improved the functional properties of soybean protein isolate, including solubility, emulsification activity, and foam stability. These changes enable uses of soybean in products like soy milk, tofu, and tofu-fortified patties. The value addition effects of radiation processing are beneficial for better utilization of soybean.
This document provides biographical information on Madhav P. Yadav including his education, work experience, professional activities, awards, and publications. It can be summarized as follows:
Yadav has a Ph.D. in Biochemistry from Southern Illinois University and has worked as a research scientist at various academic and government institutions. He has over 70 publications and has received awards for his research focusing on carbohydrate chemistry and the development of value-added products from agricultural residues.
This document discusses medium optimization and antioxidant activity of exopolysaccharide produced by Bacillus subtilis. Bacillus subtilis was isolated from soil and used to produce exopolysaccharide. Statistical methods like Plackett-Burman design and response surface methodology using central composite design were used to optimize the culture medium. The optimized medium contained specific concentrations of cane molasses, yeast extract, NaCl, and CaCl2, which produced 4.92 g/L of exopolysaccharide. The exopolysaccharide showed antioxidant activity, with a 61.19% reduction of DPPH free radicals, greater than vitamin C. The biopolymer could have applications in industries and pharmaceuticals.
This study aimed to isolate thermostable cellulases from woodland soil samples using metagenomic approaches. Genomic DNA was extracted from soil samples and used to amplify fragments of endo-β-1,4-glucanases. Twenty-five different cellulase sequences were identified, with nine cloned and tested for cellulolytic activity at 60°C. One sequence, CelMS6, displayed the highest activity at 60°C, making it a good candidate for applications requiring cellulose utilization at high temperatures.
Enhanced endoglucanase production by Bacillus aerius on mixed lignocellulosic...Mushafau Adebayo Oke
Oke, M. A., Annuar, M. S. M., and Simarani, K. (2016). "Enhanced endoglucanase production by Bacillus aerius on mixed lignocellulosic substrates." BioResources, 11(3), 5854-5869.
Isolation and purification of peroxidase from soyabeanPooja Walke
Peroxidase (EC. 1.11.1.7), an oxidoreductase, has iron porphyrin ring generally and catalyzes a redox reaction between H202 as an electron acceptor and many kinds of substrates by means of oxygen liberation from HzOz (Brill, 1996).
Production of Amphiphilic Surfactant Molecule From Saccharomyces Cerevisiae M...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.
Lipase catalyzed process for biodiesel production protein engineering and lip...Melike Şeyma Kadayıfçı
This review document discusses the use of lipases in biodiesel production. It covers lipase engineering approaches like directed evolution and rational design to improve properties such as thermostability and activity. It also discusses lipase production methods like host strain selection and metabolic engineering. The goal is to develop high-performance lipases through these techniques to enable large-scale commercialization of lipase-catalyzed biodiesel production as an alternative to fossil fuels.
This document discusses optimizing the production of exopolysaccharide (EPS) by Bacillus subtilis using various carbon and nitrogen sources. Sucrose at 2% concentration produced the highest yield of EPS. Cane molasses and rice bran were also tested as carbon sources, with cane molasses at 2% giving the highest yield. Different solvents were tested for their ability to precipitate EPS, with ethanol, diethyl ether, and methanol being effective. Fourier transform infrared spectroscopy analysis revealed the extracted polymer was composed of sucrose units. In conclusion, agro-wastes like cane molasses could be used as alternative carbon sources for the economical production of EPS.
This document summarizes several research studies:
1. One study investigated the metabolic flexibility of Pseudomonas taiwanensis VLB120 by knocking out NADH dehydrogenases and performing multi-omics analysis. It revealed the bacterium's high robustness and proposed a metabolic response to type-2 NADH dehydrogenase deficiency.
2. Another characterized the BorB thioesterase from Streptomyces parvulus, determining its structure provides a substrate binding site and specificity.
3. A third engineered Pseudomonas putida to produce bisdemethoxycurcumin by leveraging its native lignin catabolism pathways.
4. A fourth studied Aspergillus niger's
Molecular Characterization of Isolated Methyl Parathion Degrading Bacteria an...IJSRD
The burning problem in present era is pesticide residue in fruits and foods. present study focus on degradation of pesticide in contest to that total 45 methyl parathion(MP) degrading bacteria were isolated from sludge of MP producing industrial waste. According to screening 21 highly efficient pesticide degraders were selected at 500ppm concentration of standard methyl parathion as a sole carbon source. All isolates were characterized by RAPD and based on the RAPD result all 21 bacterial isolates were grouped in to 2 main clusters with 58% similarity. Characterization of MP degrading gene was carried out by using specific primer. Out of 21 isolates, 14 isolates were containing mpd gene and 4 isolates were containing opd gene while remaining 3 isolates which did not show amplification with opd/ mpd primer so it may contain other unreported gene responsible for methyl parathion degradation. Isolates were identified based on 16s rRNA sequence and one highly efficient bacterium selected for genome characterization.
1. The study characterized an apple polyphenol extract standardized to 5% phloridzin and evaluated its effects on glucose transport.
2. In vitro, the extract showed a dose-dependent inhibition of up to 60% on glucose uptake by cells, comparable to known inhibitors.
3. In mice, the extract had a lower acute toxicity than green tea extract when administered orally or intraperitoneally.
4. The results suggest the extract may help regulate blood glucose levels and have potential benefits for managing diabetes and body weight control through inhibition of intestinal glucose transport. Further human studies are needed to confirm these effects.
Recent advances in technoeconomic analysis (TEA) were reviewed:
- TEA is useful for process design, cost estimation, and identifying bottlenecks early in research.
- Studies now enable faster iteration, robust uncertainty analysis, and open-source platforms.
- Trends include more expansive system boundaries and potential integration with high-throughput experiments.
Typha grass is an invasive wetland plant with high annual productivity that can be used for biorefinery. It contains high levels of sugars and low lignin, making it suitable for conversion processes. The objective is to explore Typha capensis's potential as a biorefinery feedstock. A two-step pretreatment was performed and yielded good enzymatic digestibility of sugars. Characterization of Typha lignin showed a higher S/G ratio, indicating easier delignification. Typha deserves consideration as a promising biomass feedstock for tropical biorefining due to its high productivity, simple pretreatment requirements, and good enzymatic hydrolysability.
This document describes a study that optimized media components for xylanase production by Aspergillus fumigatus using solid state fermentation. Key findings:
1. Plackett-Burman design identified three significant nutrients (NaNO3, (NH4)2SO4, KH2PO4) influencing xylanase production.
2. Central composite design and response surface methodology were used to further analyze the effects of these nutrients and establish a regression model.
3. Under optimized conditions found by response surface analysis (NaNO3 = 0.29066 g/gds, (NH4)2SO4 = 0.14448 g/gds, KH2PO4 0
Hydrolytic activity of amylase produced in solid state fermentation by a lo...Samuel Kwatia
This document summarizes research on the production and characterization of amylase enzyme from a strain of Aspergillus niger using solid-state fermentation. Key findings include:
- Amylase was produced using optimized fermentation conditions and showed highest activity at pH 4-8 and temperatures of 30-60°C.
- Partial purification using ammonium sulfate increased the specific activity and concentration of amylase.
- The purified enzyme was able to hydrolyze starches from various sources like maize, yam and cassava, producing reducing sugars.
- Different starch sources showed varying susceptibility to hydrolysis, with maize starch being most easily broken down.
A comparative study of biopolymers and alum in the separation and recovery of...Soumyadeep Mukherjee
This study compares the effectiveness of various biopolymers and alum in recovering pulp fibers from paper mill wastewater through flocculation. Guar gum was found to be the most effective biopolymer, recovering 3.86 mg/L of fibers. Alum displayed the highest particle removal rate. Guar gum, alum, xanthan gum, and locust bean gum removed 94.68%, 97.46%, 92.39%, and 92.46% of turbidity, respectively, at a settling velocity of 0.5 cm/min. Response surface methodology was used to optimize conditions like pH, dose, and mixing speed to obtain the lowest sludge volume index for guar gum. The results indicate similar performance
Pomelo peels as alternative substrate for extracellular pectinase production ...Haritharan Weloosamy
This document summarizes a study that investigated using pomelo peels as an alternative substrate for producing pectinase enzymes through solid state fermentation with the fungus Aspergillus niger HFM-8. Initial experiments found the highest pectinase production and fungal growth on day 8 of cultivation. Various culture parameters were then optimized, finding that a substrate particle size of 0.75 mm produced the highest pectinase levels and fungal growth. After improving the fermentation parameters, pectinase production increased by 48.82% compared to initial levels, demonstrating that pomelo peels are a suitable substrate for pectinase production.
Effects of pretreatment of single and mixed lignocellulosic substrates on pro...Mushafau Adebayo Oke
A mixed substrate (MS) comprising oil palm empty fruit bunch (EFB), oil palm frond (OPF), and rice husk (RH) was evaluated for endoglucanase production by Bacillus aerius S5.2. Effects of sulphuric acid, sodium hydroxide, N-methylmorpholine-N-oxide (NMMO), and hydrothermal pretreatments on endoglucanase production were investigated. Endoglucanase production by B. aerius on the untreated (0.677 U/mL) and pretreated MS (0.305 – 0.630 U/mL) was generally similar, except that the acid (0.305 U/mL) and hydrothermal (0.549 U/mL) pretreatments that were more severe consequently produced significantly lower titres. Alkali pretreatment supported the highest enzyme production (0.630 U/mL) among all pretreatments that were studied. When endoglucanase production on the alkali-pretreated MS and single substrates (SS) was compared, alkali-pretreated EFB produced a titre (0.655 U/mL) similar to the MS, and this was significantly higher than titres recorded on OPF (0.504 U/mL) and RH (0.525 U/mL). Lower enzyme production was found to be consistent with higher pretreatment severity and greater removal of amorphous regions in all the pretreatments. Furthermore, combining the SS showed no adverse effects on endoglucanase production.
A study with enzymatic membrane reactor for conversion of lactose in to galac...Pallavi Kumari
The formation of galacto-oligosaccharides (GOS) from lactose by commercially available Biolacta FN5 (β- galactosidase, EC 3.2.1.23) derived from Bacillus circulans was studied under immobilized enzyme condition. The present work utilizes hydrophobic membrane (0.22 m pore size) for immobilization of enzyme. Experiments were conducted in a three compartment cell. The middle compartment (~25 mL) being separated by immobilized membranes was utilized for feed lactose solution; whereas, adjacent compartments were filled with distilled water. The reacted mixture solution was analyzed for tri-, tetra- and penta- forms of GOS which depended on varying amounts of initial lactose (ILC) and enzyme concentrations. Total GOS formation increased from 7 to 28% for ILC from 50 to 200 g/L. However, tri-saccharide was the major (67%) in comparison to tetra (27%) and penta (6%) forms of GOS. There was marginal difference of GOS formations while comparing the result (GOS yield) under both free (~30%) and immobilized (~28%) conditions.
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.
Production and Purification of Amylase from Bacillus subtilis Isolated from SoilDr. Amarjeet Singh
In spite of progress in biotechnology and
enzymology, the enzymes have been industrialized in recent
years for the mounting up the product development in
various arena. The ultimate goal of this study comprises the
production and purification the amylase enzyme from the
bacterial strain. A powerful amylase producer, Bacillus
subtilis ISOLATE-4 was isolated, screened and identified
from the soil sample. In order to produce extracellular
amylase, various physico-chemical parameters were
optimized. During optimization, the maximal production of
amylase by the isolate at 48 hrs of incubation in 100 rpm was
found to be 6.93U/ml, 5.94U/ml, 6.0U/ml at 45ºC, pH 6 with
1% substrate concentration respectively. Ammonium
sulphate fractionation was done for rapid precipitation of the
amylase at a concentration of 60% and exposed to dialysis
showed the 25% purification fold of an enzyme. The dialyzed
product was further subjected to DEAE-Cellulose column
chromatography resulted in an increase up to 75%
purification fold than crude enzyme. The amylase enzyme
might be suitable for the liquefaction of starch, detergent,
textile and several additional industrial applications.
Cod Removal Of An Industrial Effluent Using Nan crystalline Ceria Synthesized...IOSRJAC
Nanocrystalline ceria (CeO2) was prepared by solution combustion method using cerium nitrate as oxidizer and citric acid as fuel. The as-formed CeO2 nanopowder was characterized by Powder X-ray diffraction, Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The nanoparticles were found to be agglomerated, fluffy and porous with a mean crystallite size of about 20 nm. The as-formed ceria nanopowder was used for the removal of chemical oxygen demand (COD) of an industrial effluent. The effect of various factors such as pH, dosage of nanopowder, stirring time and sedimentation time was studied. It was found that more than 89% removal of COD could be achieved at a pH of 4, for a catalyst dosage of 0.8g of the nanopowder per liter of the industrial effluent with a sedimentation time of about 80 minutes.
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.
The papers for publication in The International Journal of Engineering& Science are selected through rigorous peer reviews to ensure originality, timeliness, relevance, and readability.
Optimization of Cultural Parameters for Cellulase Enzyme Production from Fung...IOSR Journals
Cellulalytic fungi synthesize cellulose enzyme for biodegradation of cellulose. This depends on various condition which include the source f isolation. This study was designed to determine the optimum condition necessary for cellulose production by fungi. Cellulose activities at different temperatures, pH and nitrogen sources by Rhizopus oryzae Aspergillus niger; A. flams, P. expansum and A. oryzae in liquid medium was studied and cellulose enzyme assay carried out by dinitrosalicylic acid method. All the fungal isolates have their highest cellulose activity at 400c except Penicillium expansum whose highest value of 1.28mg/ml was obtained at 320c. Cellulase produced 6m was found to be highest in all the isolate at pH 4.0 exception P expansum which occur at pH 5.5 (1.21mg/ml). The highest value e1.45mg/ml was obtained in A niger. Highest cellulose activity for A. niger, A. oryzae & P. expansum occurred in peptone. The study shows the need to determine the best physiological condition that allow for the optimal cellulose activity of fungal isolate. This will enhance their enzyme production.
This document summarizes a study on the production of bioethanol from potato and carambola juice using molds and agaricus as sources of amylase enzymes. The amylase activity of molds and agaricus was investigated under varying conditions of starch concentration, pH, incubation time, and temperature. Maximum amylase activity of 173-178 U/g was obtained for molds using 1.5% starch solution at pH 5.0 and 60°C for 30 minutes. For agaricus, highest amylase production of 14-16 U/g occurred with 1.5% starch solution at pH 6.0 and 75°C for 30 minutes. Reducing sugars were produced by fermenting potato
This document summarizes a study that characterized tannase production by Lactococcus lactis subsp lactis and its potential to enhance the nutritional value of composite sourdough. Key findings include:
1) L. lactis was able to tolerate tannin concentrations up to 0.25 mM and degrade 44.2% of tannins at 0.08 mM after 6 hours. Tannase production was inducible and the enzyme was detected intracellularly and extracellularly.
2) In a composite sourdough containing Vigna mungo flour and wheat flour, fermentation with L. lactis resulted in complete reduction of tannins (100%) and significant reduction of saponins
1 ijfaf oct-2017-4-the characteristics of rumenAI Publications
The ruminant animal feed ingredients consisting of forage with the combination of grass and legume which were almost limited in availability, the cocoa leaves as agricultural wastes or wastes containing low nutrient plantation low used as alternative feed ingredients. To overcome these problems required a simple technology that is fermentation using microorganisms indigenus so that the effluent quality cocoa can be improved. Indigenus microorganisms were microbes that exist in individuals without disturbing the activities. The purpose of this research was to improve the quality of rumen fluid characteristics and digestibility of cocoa leaves fermented by microorganisms indigenus. The method used in this study was completely randomized design factorial with three replications, treatment factors consist of A as the level of microbes (A1 = 5%, A2 = 10%, A3 = 15%) and factor B as the duration of fermentation (B1 = 2 day, B2 = 4 days, B3 = 6 days) and the variables; the nutrient content and digestibility of cocoa leaves before and after being fermented and the characteristics of rumen fluid in vitro methods. The results showed that the increasing levels of use of microbes and fermentation time can enhance the quality of cocoa leaves that reduce the content of dry material from 31.389 to 25.488% and increase the protein content of the coarse 6.153 to 7.483%. Dry Material digestibility increased from 27.379 to 32.942% and 27.432% Protein 21,435- rough. Rumen fluid characteristics were rumen fluid pH ranges from 7.427 to 7.603, NH3 production ranges from 16.820 - 19,975mg / 100ml, and Volatyl Fatty Acid production from 83.667 to 101.000 mM. The conclusion that increasing the digestibility quality cocoa fruit leather could be made by fermentation by microorganisms indigenus.
Improved Sugar Yield for Bioethanol Production by Modelling Enzymatic Hydroly...theijes
Alternative lignocellulosic substrates to produce high value-added products such as biofuel have been attractive. A Box-Behnken design was used to evaluate the effects of three parameters namely L/S ratio (50_ 100 mL/g), cellulase concentration (10_ 60 U/g) and incubation time (4_ 44h), on the enzymatic hydrolysis yield of physically pretreated Peganum harmala leaves. The fitted mathematical model allowed us to plot response surfaces as well as isoresponse curves and to determine optimal saccharification conditions. Statistical results indicated that the hydrolysis time and the enzyme concentration were the main factors influencing the release of reducing sugars. The selected optimal saccharification conditions were: L/S ratio of 75.0 mL/g, enzyme concentration of 35.0 U/g, and reaction time of 44.0h. These conditions allowed 39.6% of enzymatic hydrolysis yield versus 37.8±2.9%, respectively for the predicted values. The saccharification efficiency using enzyme treated biomass under optimized conditions was about 20-fold higher than before optimization. Fermentation of optimized cellulosic hydrolysate containing 12.6% glucose was performed using Saccharomyces cerevisiae yielded 4.75% ethanol production within 48h. These results showed a promising future of applying Peganum harmala leaves as potential lignocellulosic biomass for second generation bioethanol production.
Analytical method validation@ american chemical society@ sudeb mandalDr Sudeb Mandal
Analytical Method Validation for the Determination of
Meptyldinocap As 2,4-Dinitrooctylphenol Metabolite in Mango
and Soil Using LC-MS/MS and Dissipation Study of the
Fungicide in Indian Mango Field Ecosystem
Acs analytical method validation for the determination of md by lc-msmsDr. Sudeb Mandal
An analytical method was developed and validated for quantifying the fungicide meptyldinocap and its metabolite 2,4-dinitrooctylphenol (2,4-DNOP) in mango and soil samples using LC-MS/MS. Mango and soil samples were extracted with an acetone-methanol-HCl solution to convert meptyldinocap to 2,4-DNOP, which was then cleaned up and quantified by LC-MS/MS. The method was validated over a range of 0.025-2 μg/g and the limit of quantification was 0.025 μg/g. Recoveries from spiked samples ranged from 93-98%
This document summarizes a study on modeling the anaerobic biodegradation of fruit, vegetable, and cooked oil waste. Laboratory experiments were conducted to determine the cumulative biogas production from digesting mixtures of these substrates. Four kinetic models (exponential, Cone, Fitzhugh, and Gompertz) were used to analyze degradation performance and estimate the rate constant (k). Statistical analysis found the models fit the observed data well. A dynamic model was also developed and provided a good interpretation of the experimental process. The results showed k values were similar for vegetable and fruit digestion but different for digestion with oil.
Induced mutational studies on saccharomyces cerevisiae for bioethanol product...eSAT Publishing House
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
Isolation and Screening of Hydrogen Producing Bacterial Strain from Sugarcane...Editor IJCATR
The aim of this study is to isolate a highly competent bacterium with potent cellulose degrading capability and a better
hydrogen producer. Soil sample from sugarcane bagasse yard was isolated, serially diluted and plated on cellulose specific nutrient
agar plate. Four colonies have been isolated in which a single colony has potent cellulose degrading ability and the highest hydrogen
productivity of 275.13 mL H2 L-1. The newly isolated bacterium was morphologically and biochemically characterized. The
molecular characterization of the bacterium was carried out using 16S rDNA sequencing and the organism was identified as
Bacilllus subtilis AuChE413. Proteomic analysis such as MALDI-TOF was carried out to differentiate the isolated Bacillus subtilis
from Bacillus thuringiensis and Bacillus amyloliquefaciens. Phylogenetic tree was constructed to analyze the evolutionary
relationship among different genus and species with the newly isolated strain.
2. 82 Kumar et al.
the protocol of Au and Chan (1986). A bacterial isolate exhibiting acetate buffer (pH 5.0). The concentrated protein was fractionated
maximum enzyme activity identified as Paenibacillus polymyxa from using affinity column (1.5 x 26 cm) having agarose matrix pre-
Microbial Type Culture Collection (MTCC), Chandigarh, India, and equilibrated with 50 mM acetate buffer (pH 5) and protein fractions of
numbered as MTCC10056 was taken up for further studies. The 1.5 ml were collected upto 40 ml. The flow rate was maintained at 1
medium used in the study unless otherwise stated was a modified ml min-1 and the protein values were monitored spectro-
mineral salt medium (Rowe et al., 1975) having mango peel or other photometrically at 280 nm. The enzyme activity of all the fractions
carbohydrate as substrate. After the completion of fermentation (7th was estimated and the fractions showing the enzyme activities
day), the culture filtrate was centrifuged at 14,000 rpm, 4oC for 20 were pooled up. Electrophoresis in 10% native-polyacrylamide
min and the supernatant was assayed for CMCase activity. Different gels and 12% sodium dodecyl sulphate-polyacrylamide gels were
carbohydrate sources and agro-waste were compared for potential carried out in the discontinuous buffer systems as described by
use as substrate for cellulase production. Laemmli and Favre (1973).
Enzyme assays were performed at 30oC for 1 hr. The Results and Discussion
CMCase activity was determined as per method of Miller (1959)
Out of thirty two microbial isolates, Paenibacillus polymyxa
using carboxy methyl cellulose as substrate. The amount of released
MTCC 10056, a bacterium isolated from degrading citrus peel,
reducing sugar was quantified using glucose as standard for
exhibited maximum enzyme activity (6.069 U mg-1). The selected
determining enzyme activity and expressed in terms of U gm-1 of
isolate was further confirmed for enzyme production ability by growing
substrate utilized. One unit of enzyme activity is defined as the amount
it on pure substrates and other agro-wastes. Among the synthetic
of enzyme required to release 1 µm equivalent of glucose minute-1 g-1 of
carbon substrate tested, maximum CMCase production was
substrate utilized. Protein concentration was estimated by method of
Lowry et al. (1951). The Michaelis-Menten equation constant (Km observed with carboxymethyl cellulose (24.5U mg-1) and minimum
and Vmax) were determined by Line Weaver-Burke plot using carboxy with glucose (4.722 U mg-1). On the other hand, among agro
methyl cellulose as substrate. Reaction was carried out at 30oC in 50 wastes, mango peel (5.7 U mg-1) was found most promising where
mM acetate buffer pH 5 at various concentrations of substrate from 0- as least production (2.6 U mg-1) was observed in banana waste
3.0%. medium (Table 1).
The effect of substrates concentration 1-10% (w/v), nitrogen The CMCase production increased with increase in
addition (0.5-1.5% w/v), incubation temperature (20-70 C) and pH substrate (mango peel) concentration upto 7% (6.3 U mg-1).
(3-8) on cellulase production using mango peel as substrate was Decrease in CMCase production was observed at 10% substrate
determined. All the experiments were conducted in triplicate and the concentration (3.7 U mg-1) which might be due to the catabolite
mean values are expressed. Temperature and pH stabilities of repression and/or inhibition due to phenolics accumulation in
CMCase were determined as per method of Au and Chan (1986). the medium. Similar repression of synthesis of cellulolytic enzymes
The culture filtrate was precipitated using ammonium sulphate at has been demonstrated in many organisms (Narasimha et al.,
20-70% saturation and was dialyzed twice against the 50 mM of 2006).
0.025 0.1
Protein 0.09
CMCase activity
0.02 0.08 CM Case activity (U ml-1)
0.07
0.015 0.06
A 280nm
0.05
0.01 0.04
0.03
0.005 0.02
0.01
0 0
0 2.5 5 7.5 10 12.5 15 17.5 20 22.5 25 27.5 30 32.5 35 37.5 40
Fraction number
Fig. 1: Elution profile of protein fractions with CMCase activity
Journal of Environmental Biology January 2012
3. Production of CMCase from Paenibacillus polymyxa 83
Table - 1: CMCase production by Paenibacillus polymyxa using different 400
y = 0.0722x + 56.164
carbohydrate sources as substrate. 350 R2 = 0.5455
Substrate Specific activity of cellulase (U mg-1) 300
Sucrose 6.94±0.30* 250
Starch 10.06±1.82*
1/V
200
Carboxymethyl cellulose 24.44±3.24*
Glucose 4.72±0.24* 150
Cotton 11.33±1.28* 100
Filter paper 8.89±1.67*
50
Cellulose 11.67±1.94*
Maltose 9.63±1.53* 0
Cellulobiose 13.98±1.34* -2000 -1000 0 1000 2000 3000 4000
Banana waste 2.59±0.45* 1/S
Bael1 fibre 3.38±1.18* Fig. 3: Lineweaver-Burke plot for determination of Km and Vmax using
Mahua2 pomace 1.67±0.92* carboxy methyl cellulose as substrate (1/V=Reciprocal of velocity, 1/S=
Mango peel 5.71±1.30* Reciprocal of substrate concentration), Km= Michaelis constant)
Aonla3 pomace 2.87±0.97*
Maize corn 5.05±0.84*
The effectiveness of nitrogen source in supporting CMCase
1 = Aegle marmelos, 2 = Madhuca indica, 3 = Emblica officinalis,
production along with growth and secretion of extracellular protein
* Values are mean of three replicates ± S.D
has also been reported in many microorganisms (Narasimha et al.,
2006). Since, mango peel has a poor protein status (2.9%) the effect
of addition of nitrogen on CMCase production was worked out. The
addition of ammonium sulphate (1.5% ) as nitrogen supplement
110 kDa
enhanced CMCase production by 28.2%.
For CMCase production, pH of 5.5 and incubation at 37oC
were found to be optimum (7.81 U mg-1). Increase or decrease in
66 kDa pH or temperature significantly affected the enzyme production. Baig
et al. (2004) have reported pH 6.0 as optimum for maximum cellulase
51 kDa production from Trichoderma lignorum using banana waste. Optimum
temperature for CMCase production was reported at 30oC by Bacillus
sp. and Pseudomonas fluorescens (Khyami-Horami 1996; Bakare
et al., 2005).
34 kDa After ammonium sulphate precipitation of culture filtrate,
30 kDa the crude protein obtained was purified to the extent of 28.24 fold
with 1.99% recovery (Table 2) by affinity chromatography on
agarose column. The elution profile of protein fractions revealed that
fractions numbering 10 to 25 showed peak CMCase activity (Fig. 1).
26.5 kDa
25 kDa Purification to extent of 24-26 folds had been reported earlier in
cellulase and pectinase (Bakare et al., 2005; Ariffin et al., 2006;
Arotupin, 2008).
The fraction exhibiting enzyme activities were pooled and
18 kDa analyzed on 12% SDS-PAGE and 10% Native PAGE. Two bands
corresponding to 34 and 26.5 kDa were observed in 12% SDS-
PAGE (Fig. 2) and it was observed as single band in 10% Native gel.
14 kDa
Earlier, 29 kDa alkaline cellulase and 30-65 kDa cellulase was reported
in Bacillus sp and Bacillus pumilus, respectively (Khyami-Horami,
1996; Ariffin et al., 2006) in denaturing PAGE; where as 60-70
kDa cellulase has been obtained in 10% Native PAGE (Giorgini,
1992). Similarly, Chen et al. (2004) has reported CMCase of 94
Fig. 2: Electrophoretic banding pattern of protein on 12% SDS-PAGE kDa in Sinorhizobium fredii while Coral et al. (2002) identified 83
(M = Marker, L1 = Purified cellulase) and 50 kDa CMCase in Aspergillus niger wild type strain Z10.
Journal of Environmental Biology January 2012
4. 84 Kumar et al.
Table - 2: Summary of CMCase purification by Paenibacillus polymyxa
Purification step for CMCase activity Total protein Specific activity Yield Fold
purification (U mg-1) (mg) (U mg-1) % purification
Culture filtrate (Crude) 24.6 140 0.18 100 1
Ammonium sulphate 10.46 14.07 0.74 42.27 4
Affinity chromatography 0.5 0.1 4.82 1.99 28.24
Since cellulase is a heteromeric multienzyme complex, these Giorgini, J.F.: Purification and partial characterization of two isozyme of
two bands might correspond to different components of cellulase cellulase from GA3-treated coffee endosperm. R. Bras. Fisiol. Veg., 4,
75-80 (1992).
enzyme complex in P. polymyxa. CMCase of P. polymyxa exhibited Hang, Y.D. and E.E. Woodams: Apple Pomace: Potential substrate for production
maximum cellulase activity at 30oC (8.4 U mg-1) and at pH-5 (9.4 U of β-Glycosidase by Aspergillus foetidus. Lebensm-Wiss. U-Technol.,
mg-1). The purified enzyme was found to be stable over range of 27, 587-589 (1994).
20-60oC, at pH 4.0-7.5, though, only 25% activity was retained at Ikrma-Ul-Haq., M.M. Javed and T.S. Khan: An innovative approach for
hyper production of cellulolytic and hemicellulolytic enzyme by
pH 7.5. Enzyme showed a temperature stability range between consortium of Aspergillus niger MSK-7 and Trichoderma viride MSK-
20-60oC. The stability declined at temperature higher than 60oC 10. Afr. J. Biotechnol., 5, 609-614 (2006).
while negligible enzymatic activity was observed at 70oC. The Km Khyami-Horami, H.: Partial purification and some properties of an alkaline
and Vmax for cellulase from P. polymyxa was 8.73 mg ml-1 and cellulase from an alkalophilic Bacillus sp. World J. Microbiol. Biotechnol.,
12, 525-529 (1996).
17.805 mM min-1, respectively (Fig. 3). Earlier, Km values for Krishna, C.: Production of bacterial cellulases by solid state bio processing of
CMCase have reported from Alternaria alternata as 16.64 mg ml-1 banana wastes. Bioresource Technol., 69, 231-239 (1999).
(Macris, 1984) and Candida peltala as 66 mg ml-1 (Saha, 1996). Laemmli, U.K. and M. Favre: Maturation of the head of bacteriophage T4. I.
Lower Km value has the advantage that the enzyme maintains DNA packaging events. J. Mol. Biol., 80, 575-599 (1973).
Lowry, O.H., N.J. Rosebrough, A.L. Farr and R.J. Randall: Protein
sufficient degradation rate which leads to better substrate
measurement with the Folin phenol reagent. J. Biol. Chem., 193, 265-275
transformation. The study revealed that mango peel may serve as (1951).
substrate for CMCase production using P. polymyxa with optimized Lynd, L.R., P.J. Weimer, W.H. van Zyl and I.S. Pretorius: Microbial cellulose
conditions. utilization: Fundamentals and biotechnology. Micro. Mol. Biol. Rev.,
66, 506-577 (2002).
Acknowledgments Macris, B.J.: Production and characterization of cellulase and β-glucosidase
from a mutant Alternaria alternate. Appl. Environ. Microbiol., 47, 560-565
Authors are thankful to the Director, Central Institute for (1984).
Subtropical Horticulture, Lucknow for his constant support. The Maurer, K.H.: Development of new cellulases, in enzymes in detergency,
research was funded by Application of Microorganism in Agriculture (Ed.: H. Jan, E. Van) (Marcel Dekker, New York). pp. 175-202 (1997).
Miller, G.L.: Use of dinitrosalycylic acid reagent for determination of sugar.
and Allied Sector (AMAAS) networking project of Indian Council of Anal. Chem., 31, 426-428 (1959).
Agricultural Research. Narasimha, G., A. Sridevi, V. Buddolla, C.M. Subhosh and R.B. Rajasekher:
Nutrient effects on production of cellulolytic enzymes by Aspergillus
References niger. Afr. J. Biotechnol., 5, 472-476 (2006).
Ariffin, H., N. Abdullah, K. Umi, Y. Shirai and M.A Hassan: Production and Nigam, P. and D. Singh: Processing of agriculture wastes in solid state
characterization by Bacillus pumilus EB3. Int. J. Eng. Technol., 3, 47-53 fermentation for cellulolytic enzyme production. J. Scientific Industrial
(2006). Research, 55, 457-463 (1996).
Arotupin, D.J., F.A. Akinyosoye and A.K. Onifade: Purification and Omojasola, P., Folakemi Jilani, O. Priscilla and S.A. Ldiyemi: Cellulase
characterization of pectinmethylesterase from Aspergillus niger isolated production by some fungi cultured on pineapple waste. Nat. Sci., 6,
from cultivated soil. Afr. J. Biotechnol., 7, 1991-1998 (2008). 64-79 (2008).
Au, K.S and K. Chan: Carboxymethyl cellulase production by Bacillus Rowe, J.J., D. Goldberg and R.E. Amelunxen: Development of defined and
substilis. Microbios, 48, 93-108 (1986). minimal media for the growth of Bacillus stereothermophilus. J. Bact.,
Baig, M.M.V., M.L.B. Baig, M.I.A. Baig and M. Yasmeen: Saccharification of 124, 279-285 (1975).
banan agro-wate by cellulolytic enzymes. Afr. J. Biotechnol., 3, Saha, B.C. and R.J. Bosthast: Production, purification and characterization of
447-450 (2004). a higher glucose tolerant novel β-glucosidase from Candida peltata.
Bakare, M.K., I.O. Adewale, A. Ajai and O.O. Shonukan: Purification and Appl. Environ. Microbiol., 62, 3165-3170 (1996).
characterization of cellulase from the wild-type and two improved Sukumaran, R.K., R.R. Singhania and A. Pandey: Microbial cellulases?
mutants of Pseudomonas fluorescens. Afr. J. Biotechnol., 4, 898-904, Production, applications and challenges. J. Sci. Indus. Res., 64,
(2005). 832-844 (2005).
Chen, P.J., T.C. Wei, Y.T. Chang and L.P. Ling: Purification and characterization Sun, H., X. Ge, Z. Hao and M. Peng: Cellulase production by Trichoderma
of carboxylmethyl cellulase by Sinorhizobium fredii. Bot. Bull. Acad. sp. on apple pomace under solid state fermentation. Afr. J. Biotechnol.,
Sin., 45, 111-118 (2004). 9, 163-166 (2010).
Coral, G., B. Arikan, M.N. Unaldi and H. Guvenmez: Some properties of Szengyel, Z., G. Zacchi, A. Varga and K. Rexzet: Cellulase production of
carboxymethyl cellulase of Aspergillus niger Z10 wild-type strain. Trichoderma reesei Rut C-30 using steam pre treated spruce hydrolytic
Turk. J. Biol., 26, 209-213 (2002). potential on different substrates. Appl. Biochem. Biotechnol., 84-86,
Garg, N. and M. Ashfaque: Mango peel as substate for production of 679-691 (2000).
extracellular polygalacturonase from Aspergillus fumigatus. Ind. J. Tandon, D.K., N. Garg and S.K. Kalra: Extraction of pectin and fibre from
Horti., 67, 140-143 (2010). mango peel. Beverage and Food World, 22, 20-21 (1995).
Journal of Environmental Biology January 2012