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.
“Production and optimization of lipase from bacillus subtillis”Pooja Walke
Lipases (try acryl glycerol acylhydrolase ) are the enzymes which catalyze the hydrolysis and the synthesis of ester formed from glycerol and long chain fatty acid.
Production of secondary metabolites : enzymes which involves the upstream technological process
Introduction
History
Process involved
Contribution of different micro-organisms
Flowchart
Example: Methods Production of Amyalse in industrial view
The following presentation is only for quick reference. I would advise you to read the theoretical aspects of the respective topic and then use this presentation for your last minute revision. I hope it helps you..!!
Mayur D. Chauhan
“Production and optimization of lipase from bacillus subtillis”Pooja Walke
Lipases (try acryl glycerol acylhydrolase ) are the enzymes which catalyze the hydrolysis and the synthesis of ester formed from glycerol and long chain fatty acid.
Production of secondary metabolites : enzymes which involves the upstream technological process
Introduction
History
Process involved
Contribution of different micro-organisms
Flowchart
Example: Methods Production of Amyalse in industrial view
The following presentation is only for quick reference. I would advise you to read the theoretical aspects of the respective topic and then use this presentation for your last minute revision. I hope it helps you..!!
Mayur D. Chauhan
Lipase production and purification Likhith KLIKHITHK1
Lipase (tri acyl glycerol acyl hydrolase, EC 3.1.1.3) catalyzes the hydrolysis of the carboxyl ester bonds in tri acyl glycerols to produce di acyl glycerols, mono acyl glycerols, fatty acids and glycerol under aqueous conditions and the synthesis of esters in organic solvents.
Under the controlled conditions, lipases are able to catalyze a large number of reactions. Lipases of microbial origin are of considerable commercial importance, because of the high versatility and high stability, moreover, the advantage of being readily produced in high yields.
Many microbial lipases have been commercially available in free or immobilized form. Numerous species of bacteria (Bacillus, Pseudomonas, and Burkholderia), yeasts (Candida rugosa, Yarrowia lipolytica, and Candida antarctica) and molds (Aspergillus, Trichoderma viride) produce lipases with different enzymological properties and specificities but microbes are known to be more potent lipase producer.
Use of microbes in industry. Production of enzymes-General consideration-Amyl...Steffi Thomas
Industrial uses of microbes, properties of useful industrial microbes, various industrial products, production of enzymes-general consideration-amylase, catalase, peroxidase, lipase, protease, penicillinase, procedure for culturing bacteria and inoculum preparation, submerged fermentation and solid state fermentation, uses of different enzymes
Screening and Production of Protease Enzyme from Marine Microorganism and Its...iosrjce
Marine sediment samples were collected from the Gulf of Mannar, Mandapam coast to screen for
protease producing microbes. Among the five isolates screened only two isolates showed maximum proteolytic
activity with the zone of 21mm and 19mm respectively. Biochemical characterization of the isolates were
performed and identified as strain P2 belonged to Bacillus subtilis and strain P5 belonged to Bacillus
licheniformis. Both the strains have the ability to tolerate 7%Nacl concentration. The amount of protease
produced was expressed in microgram of tyrosine released under standard assay conditions. The total protein
content of crude enzyme extracts of Bacillus subtilis and Bacillus licheniformis were quantified which revealed
21.2mg/ml for strain P2 and 22.4mg/ml of protein content was presented by strain P5. The proteolytic bacteria
gave an optimum performance were both strains exhibited the enzymes stable at PH
7. In the present study
Bacillus subtilis showed a remarkable activity at 40ºC where as Bacillus licheniformis exhibited maximum
activity at 50ºC. Studies pertaining to carbon sources starch and lactose were utilized by Bacillus subtilis and
Bacillus licheniformis and maximum production was achieved. Among the different nitrogen sources tested
yeast extract induced maximum proteolytic activity where as ammonium sulphate was found to be the best
nitrogen sources for protease production. The crude enzyme was efficient to remove hair dye and blood stain by
Bacillus subtilis and Bacillus licheniformis
It describes the history, production, and substrates used in the production of the enzyme. also, emphasize the application of amylase in food industry.
Amylase is an enzyme that catalyzes the hydrolysis of starch into sugars.
Amylase is present in the saliva of humans and some other mammals, where it begins the chemical process of digestion. Foods that contain large amount of starch but less amount of sugar, such as rice and potatoes, may acquire a slightly sweet taste as they are chewed because amylase degrades some of their starch into sugar.
α- amylase is a protein enzyme that hydrolyses alpha bonds of large, alpha - linked polysaccharides, such as starch and glycogen, yielding glucose and maltose. It is a major form of amylase found in humans and other mammals.
Many of the enzymes used in the industries are extracellular derived from microorganisms. Among various extracellular enzymes, alpha amylase ranks first in terms of commercial exploitation.
Bacteria and fungi secrets amylases to the outside of the cells to carryout extracellular digestions when they have broken down the soluble starch, the soluble end products such as Glucose or Maltose are absorbed into their cells.
The industrially important Bacillus strains which are extensively used to produce alpha amylase, are, B. licheniformis, B. subtilis etc. B. amyloliquefaciens
Bacillus licheniformis is a Gram-positive endospore forming organism that can be isolated from soils and plant material all over the world.
This organism is used extensively for large-scale industrial production of exoenzymes as it can secrete large quantities of proteins of up to 20–25 g/l.
The use of the submerged culture is advantageous because of the ease of sterilization and its process control.
The objective of this work was to study the pattern and the comparison of α-amylase production by using two strains of Bacillus licheniformis, MTCC 2617 and MTCC 2618 using four different substrates starch, rice, wheat and ragi powder as carbon source.
Ragi or finger millet is round, soft yet firm and rich brown in color. It is probably the only edible solid you are advised to swallow not chew. A gram of ragi has 72% carbohydrate, 3.6% fiber, 7.3% of protein, vitamin B and a good combination of minerals.
Alkaline Protease - One of the class of protease enzyme.
An extracellular enzyme.
Performs proteolysis, that is, protein catabolism by hydrolysis of the peptide bonds.
Active at alkaline pH 8 to 12 and at temperature 30⁰-80⁰C.
Molecular weight is about 20,000 to 45,000 Dalton.
The structure is determined by X-ray crystallography.
EC (Enzyme Commission) Number: 3.4.21–24.99
In 1971, Japanese scientist Koki Horikoshi first reported the production of alkaline protease from bacteria.
How to produce enzyme based products at home: cleaning & Personal careMurray Hunter
The production of enzyme based products in Thailand & emerging cosmetic & personal care industry. Presented to the IAB WOMEN IN SCIENCE INTERNATIONAL SYMPOSIUM ON
THE SCIENCE OF HEALTH, BEAUTY AND AGEING
7-8 MAY 2012
PRODUCTION OF PROTEASE BY ALKALOPHILIC BACILLUSSUBTILIS IN BIOREACTOR AND ITS...AM Publications
The current studies were aimed at to investigate role of pH, dissolved oxygen for production protease in bioreactor by alkalophilic bacterium and application of saw dust for its purification. The production of proteolytic enzyme by Bacillus subtilis IC-5 started as pH of medium falls to 9 and reached to maximum at pH 7 i.e., 4400 Uml-1 . Likewise dissolved oxygen decreased in the medium as the protease production progresses. Saw dust was successively utilized for partial purification of protease. The partial purification of protease increased the specific activity to5.3 fold. The optimum pH and temperature for purified activity was 11 and 700C, respectively. The purified enzyme was stable up to pH 12 and 80oC.
UNIT-5 Protein Engineering: Brief introduction to protein engineering,Use of ...Shyam Bass
UNIT-5 6th Sem B.PHARMA PHARMACEUTICAL BIOTECHNOLOGY)
Protein Engineering: Brief introduction to protein engineering, Use of microbes in industry, Production of enzymes-general considerations, Amylase, Catalase, peroxidase, Lipase Basic principles of genetic engineering
BY- SHYAM BASS
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).
In the field of biotechnology there are many industrial applications that result in biotech products that we use everyday at home. Some of these are food science applications that utilize enzymes to produce or make improvements in the quality of different foods. In the dairy industry, some enzymes are required for the production of cheeses, yogurt and other dairy products, while others are used in a more specialized fashion to improve texture or flavour.
Isolation and Purification of Enzymes
Enzymes are unstable molecules with a definite physico chemical organization. Even a slight change in this organization reduces the activity of enzyme and sometimes the enzyme is totally inactivated.
Therefore, the enzymes have to be isolated under controlled conditions of pH, ionic strength and temperature. Since they are proteinaceous in nature, standard extraction and purification procedures for enzymes are the same as those used for proteins except that the activity of the enzyme is assayed at each of the following four steps of extraction and purification.
Purification of Enzymes - Enzyme purification involves three steps, electrophoresis. These three techniques described in the following text
1.Dialysis
2.Chromatography.
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
Lipase production and purification Likhith KLIKHITHK1
Lipase (tri acyl glycerol acyl hydrolase, EC 3.1.1.3) catalyzes the hydrolysis of the carboxyl ester bonds in tri acyl glycerols to produce di acyl glycerols, mono acyl glycerols, fatty acids and glycerol under aqueous conditions and the synthesis of esters in organic solvents.
Under the controlled conditions, lipases are able to catalyze a large number of reactions. Lipases of microbial origin are of considerable commercial importance, because of the high versatility and high stability, moreover, the advantage of being readily produced in high yields.
Many microbial lipases have been commercially available in free or immobilized form. Numerous species of bacteria (Bacillus, Pseudomonas, and Burkholderia), yeasts (Candida rugosa, Yarrowia lipolytica, and Candida antarctica) and molds (Aspergillus, Trichoderma viride) produce lipases with different enzymological properties and specificities but microbes are known to be more potent lipase producer.
Use of microbes in industry. Production of enzymes-General consideration-Amyl...Steffi Thomas
Industrial uses of microbes, properties of useful industrial microbes, various industrial products, production of enzymes-general consideration-amylase, catalase, peroxidase, lipase, protease, penicillinase, procedure for culturing bacteria and inoculum preparation, submerged fermentation and solid state fermentation, uses of different enzymes
Screening and Production of Protease Enzyme from Marine Microorganism and Its...iosrjce
Marine sediment samples were collected from the Gulf of Mannar, Mandapam coast to screen for
protease producing microbes. Among the five isolates screened only two isolates showed maximum proteolytic
activity with the zone of 21mm and 19mm respectively. Biochemical characterization of the isolates were
performed and identified as strain P2 belonged to Bacillus subtilis and strain P5 belonged to Bacillus
licheniformis. Both the strains have the ability to tolerate 7%Nacl concentration. The amount of protease
produced was expressed in microgram of tyrosine released under standard assay conditions. The total protein
content of crude enzyme extracts of Bacillus subtilis and Bacillus licheniformis were quantified which revealed
21.2mg/ml for strain P2 and 22.4mg/ml of protein content was presented by strain P5. The proteolytic bacteria
gave an optimum performance were both strains exhibited the enzymes stable at PH
7. In the present study
Bacillus subtilis showed a remarkable activity at 40ºC where as Bacillus licheniformis exhibited maximum
activity at 50ºC. Studies pertaining to carbon sources starch and lactose were utilized by Bacillus subtilis and
Bacillus licheniformis and maximum production was achieved. Among the different nitrogen sources tested
yeast extract induced maximum proteolytic activity where as ammonium sulphate was found to be the best
nitrogen sources for protease production. The crude enzyme was efficient to remove hair dye and blood stain by
Bacillus subtilis and Bacillus licheniformis
It describes the history, production, and substrates used in the production of the enzyme. also, emphasize the application of amylase in food industry.
Amylase is an enzyme that catalyzes the hydrolysis of starch into sugars.
Amylase is present in the saliva of humans and some other mammals, where it begins the chemical process of digestion. Foods that contain large amount of starch but less amount of sugar, such as rice and potatoes, may acquire a slightly sweet taste as they are chewed because amylase degrades some of their starch into sugar.
α- amylase is a protein enzyme that hydrolyses alpha bonds of large, alpha - linked polysaccharides, such as starch and glycogen, yielding glucose and maltose. It is a major form of amylase found in humans and other mammals.
Many of the enzymes used in the industries are extracellular derived from microorganisms. Among various extracellular enzymes, alpha amylase ranks first in terms of commercial exploitation.
Bacteria and fungi secrets amylases to the outside of the cells to carryout extracellular digestions when they have broken down the soluble starch, the soluble end products such as Glucose or Maltose are absorbed into their cells.
The industrially important Bacillus strains which are extensively used to produce alpha amylase, are, B. licheniformis, B. subtilis etc. B. amyloliquefaciens
Bacillus licheniformis is a Gram-positive endospore forming organism that can be isolated from soils and plant material all over the world.
This organism is used extensively for large-scale industrial production of exoenzymes as it can secrete large quantities of proteins of up to 20–25 g/l.
The use of the submerged culture is advantageous because of the ease of sterilization and its process control.
The objective of this work was to study the pattern and the comparison of α-amylase production by using two strains of Bacillus licheniformis, MTCC 2617 and MTCC 2618 using four different substrates starch, rice, wheat and ragi powder as carbon source.
Ragi or finger millet is round, soft yet firm and rich brown in color. It is probably the only edible solid you are advised to swallow not chew. A gram of ragi has 72% carbohydrate, 3.6% fiber, 7.3% of protein, vitamin B and a good combination of minerals.
Alkaline Protease - One of the class of protease enzyme.
An extracellular enzyme.
Performs proteolysis, that is, protein catabolism by hydrolysis of the peptide bonds.
Active at alkaline pH 8 to 12 and at temperature 30⁰-80⁰C.
Molecular weight is about 20,000 to 45,000 Dalton.
The structure is determined by X-ray crystallography.
EC (Enzyme Commission) Number: 3.4.21–24.99
In 1971, Japanese scientist Koki Horikoshi first reported the production of alkaline protease from bacteria.
How to produce enzyme based products at home: cleaning & Personal careMurray Hunter
The production of enzyme based products in Thailand & emerging cosmetic & personal care industry. Presented to the IAB WOMEN IN SCIENCE INTERNATIONAL SYMPOSIUM ON
THE SCIENCE OF HEALTH, BEAUTY AND AGEING
7-8 MAY 2012
PRODUCTION OF PROTEASE BY ALKALOPHILIC BACILLUSSUBTILIS IN BIOREACTOR AND ITS...AM Publications
The current studies were aimed at to investigate role of pH, dissolved oxygen for production protease in bioreactor by alkalophilic bacterium and application of saw dust for its purification. The production of proteolytic enzyme by Bacillus subtilis IC-5 started as pH of medium falls to 9 and reached to maximum at pH 7 i.e., 4400 Uml-1 . Likewise dissolved oxygen decreased in the medium as the protease production progresses. Saw dust was successively utilized for partial purification of protease. The partial purification of protease increased the specific activity to5.3 fold. The optimum pH and temperature for purified activity was 11 and 700C, respectively. The purified enzyme was stable up to pH 12 and 80oC.
UNIT-5 Protein Engineering: Brief introduction to protein engineering,Use of ...Shyam Bass
UNIT-5 6th Sem B.PHARMA PHARMACEUTICAL BIOTECHNOLOGY)
Protein Engineering: Brief introduction to protein engineering, Use of microbes in industry, Production of enzymes-general considerations, Amylase, Catalase, peroxidase, Lipase Basic principles of genetic engineering
BY- SHYAM BASS
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).
In the field of biotechnology there are many industrial applications that result in biotech products that we use everyday at home. Some of these are food science applications that utilize enzymes to produce or make improvements in the quality of different foods. In the dairy industry, some enzymes are required for the production of cheeses, yogurt and other dairy products, while others are used in a more specialized fashion to improve texture or flavour.
Isolation and Purification of Enzymes
Enzymes are unstable molecules with a definite physico chemical organization. Even a slight change in this organization reduces the activity of enzyme and sometimes the enzyme is totally inactivated.
Therefore, the enzymes have to be isolated under controlled conditions of pH, ionic strength and temperature. Since they are proteinaceous in nature, standard extraction and purification procedures for enzymes are the same as those used for proteins except that the activity of the enzyme is assayed at each of the following four steps of extraction and purification.
Purification of Enzymes - Enzyme purification involves three steps, electrophoresis. These three techniques described in the following text
1.Dialysis
2.Chromatography.
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
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.
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
A comparative study on classification of image segmentation methods with a fo...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
Performance of nano crystalline h zsm-5 as additive in fcc catalyst a revieweSAT 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
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.
Hyperspectral image mixed noise reduction based on improved k svd algorithmeSAT 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.
The interconnecting mechanism for monitoring regular domestic conditioneSAT 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
Secure and efficient key pre distribution schemes for wsn using combinatorial...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
Experimental studies on pressure drop in a sinusoidal plate heat exchanger ef...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
Effect of zeolite types ltx and lta on physicochemical parameters of drinking...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
Design and implementation of address generator for wi max deinterleaver on fpgaeSAT 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.
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
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.
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.
Simulation of pedestrian at intersection in urban congested areaeSAT 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
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
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 antimicrobial activity of rhamnolipid (biosurfactant) from oil ...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 Characterization and Screening of fungal Lipase from oil contaminat...AI Publications
Present scenario demands a more sustainable, ecofriendly and economic measures globally to deal with the growing problems of environmental issues. The main goal of this work is to opt for such ideas and technologies which involve cleaner and greener procedures for utilizing waste materials for deriving value added products. The soil pertaining to the areas of oil mills contains densely population of various microbes’, especially fungal origin. These microbes are rich in lipase content (due to oil source). Thus in this we isolated fungal colonies from this oil rich soil, cultured in laboratory, fermented them under various conditions to extract fungal enzyme i.e. lipase and then used it for further applications. Lipases are highly versatile and industrially important enzymes. Deriving the lipases from waste soil is the main attraction of this work and is a venture strategizing the “best from waste” approach.
Microbial Production Of Alkaline Proteases And Evaluation Of Its Performances...Shafkat Shamim Rahman
A high alkaline protease producing bacterial strain was isolated and identified a local soil sample. The organism was gram positive and forms spore during adverse condition in the growth medium. After various tests it was suggested and the features agreed with the description of Bacillus subtilis. It was also identified as B. subtilis with 99.9% identity by API 50 CHB. The enzyme hydrolyses a number of proteins including azocasein which suggests that it is an extracellular alkaline protease. The experimentally determined isoelectric point was 5.1 and the optimal enzyme activity was at 60°C and at pH 8.5. The esterase preferentially hydrolyzed short-chain fatty acids. Native enzyme preparations typically showed a Michaelis constant (Km) and Vmax of 0.40mM and 12,200 U mg)-1, respectively. This microbial enzyme was partially purified by ammonium sulfate fractionation, dialysis, DEAE cellulose chromatography and electrophoretic analysis. Enzyme purity was tested by SDS-PAGE. Quantitative estimation has shown that 40mL of culture supernatant could dehair 2×1 cm of leather completely in 9 hours. In future the tanneries will use a combination of chemical and enzymatic processes. In practical applications, protease is a useful enzyme for promoting the hydrolysis of proteins and showing significant industrial applications.
ABSTRACT- Biosurfactant is a structurally diverse group of surface-active molecule, synthesized by microorganisms. Kocuria rosea and Pseudomonas aeruginosa strains isolated from pesticide contaminated soil, which produces biosurfactant were studied. Curd whey was used as a cheap source of growth medium for biosurfactant production. There was formation of stable emulsions of biosurfactant containing broth with vegetable oil and kerosene. These strains produced a clear zone in oil spreading test, which is an indicative of the good biosurfactant activity. Both the strains produced extra cellular biosurfactant in the culture media and showed good foam stability in the culture medium. Biosurfactant was efficiently extracted from the culture broth by acetone-HCl precipitation. The biosurfactants from the two species, namely Kocuria rosea and Pseudomonas aeruginosa were found to have no effects on germinating seedlings of Glycine max, Pisum sativum and Spinacia oleracea, when treated with 25%, 50%, 75% and 100% with the combination of curd whey in the making of 100ml volume. Curd whey as a control was taken with no surfactant. Our study suggested an efficient use in surfactant aided bioremediation in agricultural land.
Key-words- Biosurfactant, Kerosene, Emulsification, Oil spreading, Kocuria rosea, Pseudomonas aeruginosa, Glycine max, Pisum sativum, Spinacia oleracea
Proteases are protein-degrading enzymes that catalyses hydrolytic reaction in which protein molecules are degraded into peptides and amino acids. Thermostable alkaline proteases are of particular great interest for industrial application because they are stable and active at temperature above 60-70˚C. Thermophiles are found in wide array of environment such as mushroom compost material, nest, hay, wood chips, grains, soil, manure, coal mines etc. Alkaline proteases are most important industrial enzymes and they occupy about 60% of total enzyme market. From the soil samples, eight different fungal species were isolated through soil dilution plate method. In the present study, two fungi Aspergillus nidulans and Aspergillus glaucus from mushroom compost and two fungi Aspergillus terrus, and Aspergillus fumigates from cow manure, showing alkaline protease activity, were isolated. The zones of clearance were observed in Aspergillus nidulans, Aspergillus glaucus, Aspergillus terrus, and Aspergillus fumigatus species of fungi isolated from cow manure and mushroom compost. The best enzyme production was observed in Aspergillus terrus (1.005 ± 0.057 IU/mg protein) obtained from cow manure and the minimum enzyme activity was observed with Aspergillus glaucus (0.278 ± 0.026 IU/mg protein). However, more studies are required to assess the potential of Aspergillus nidulans, Aspergillus glaucus, Aspergillus terrus, and Aspergillus fumigatus species. Key-words- Alkaline protease, Thermophiles, Zone of clearance, Trichloroacetic acid
Bacterial pigments have many applications in current day to day life. The pigments produced by chromobacteria can be used for various applications like dairy, pharmaceutical, and food etc. In this study, three types of pigments were isolated i.e. yellow from Xanthomonas sp., pinkish Red from Rhodotorula sp., and orange from Sarcina sp. Pigmented bacterial isolates were obtained from the soil samples and used for the pigment extraction study. We studied that the pigment producing bacteria and identified the color producing pigments. Soil samples from Pondicherry, Cuddalore, Chennai, and Andhra sea coast were collected and used for isolation of microbes producing pigments. Purification of extracted pigments were done by column chromatography, whereas identification and characterization of purified pigment done by UV-Visible spectrophotometry and GC/MS analysis etc. The pigment isolated from bacterial sp. were used for the antimicrobial activity, antioxidant, and anticancer & transformation studies. The bacterial extracts of carotenoid pigment extracted and used as natural colorants for food products and dying of cloth.
Key-words: - Soil samples, GC/MS analysis, UV-Visible spectrophotometry, Carotenoid, Pigment extraction
Optimization of process parameters for l asparaginase production by aspergill...eSAT Journals
Abstract L-asparaginase (L-asparagine amido hydrolase, E.C.3.5.1.1) is an extra cellular enzyme that has received considerable attention since it is used as an anticancer agent. L-asparaginase belongs to an amidase group that hydrolyses the amide bond in L-asparagine to aspartic acid and ammonia. The clinical action of this enzyme as an anti-carcinogenic is attributed to the reduction of L-asparagine; tumour cells unable to synthesise this amino acid are selectively killed by L-asparagine deprivation. L-Asparaginase has its application in food industry also. It helps in reducing the content of acrylamide in baked food products by hydrolysing the L-asparagine. L-Asparaginase is majorly produced by microorganisms including bacteria, yeast and fungi. The potential of Aspergillus terreus MTCC 1782 using cauliflower stalk: corn ears (3.75: 1.25) as substrate under SSF is the purpose of the study. Solid state fermentation (SSF) is a very effective technique opposed to submerged fermentation in various aspects. Various fermentation parameters such as types of agro material, their ratios, carbon source, nitrogen source, inoculum level, moisture content, temperature, pH, fermentation time, metal salts, and L-asparagine concentration, which influence the rate of enzyme production under SSF, were optimized. The optimized production of L-asparaginase has been obtained at 35°C for 4 days with a pH of 9.0, along with 50% moisture content, and 20% inoculum volume as the optimized fermentation conditions. The optimization was done using a ‘one-factor-at-a-time’ approach. The highest yield was obtained with, sucrose (1%w/v), ammonium sulphate (1%w/v), NaCl (1%w/v), L-asparagine (1%w/w), added to the fermentation medium, as supplements. Use of cauliflower stalk along with corn ear as potential raw materials for enzyme production could be of great commercial significance. Keywords: L-asparaginase, chemotherapeutic agent, Aspergillus terreus, SSF, mixed substrate, optimization
Isolation, characterization of aspergillus fumigatus and optimization of cult...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, characterization of aspergillus fumigatus and optimization of cult...eSAT Journals
Abstract The soil samples were collected from different depths of paddy and sugarcane fields. The samples were primarily screened for isolation of amylase producing fungi. Among the isolated fungi, amylase producing isolates were identified by growing on starch agar media. The isolate (15F) which form the maximum zone of clearance on starch agar media by iodine was identified and it was subcultured on potato dextrose agar (PDA). The isolate (15F) was morphologically characterized by performing cotton blue staining and scanning electron microscopic observations under required magnifications. Molecular characterization of isolate (15F) was performed by ITS/5.8S rRNA and β-tubulin gene sequence analysis and it was confirmed as Aspergillus fumigatus (MTCC Acc No 11399). Optimization of cultural conditions for maximum production of amylase was carried out by different cereal flours, incubation periods, temperatures, nitrogen sources and with different phosphate concentrations. Aspergillus fumigatus showed maximum amylase activity (230±0.7U/mg protein) when cultured in finger millet at 350C for 72hrs of incubation period. Keywords: Amylase, Aspergillus fumigatus, cereal flour, submerged fermentation
The Role of Cell Wall-Degrading Enzymes in the Development of Anthracnose Dis...Agriculture Journal IJOEAR
— The ability of Colletotrichumtruncatum CP2 in producing pectinolytic and cellulolytic enzymes was evaluated by shake flask fermentations. The results of enzymatic activity experiment indicated that PG was the first cell wall-degrading enzymes detected and the activities obtained were higher (0.24±0.10 U/mL) than other enzymes, which appeared later and in lower amount. After the cell wall was degraded by the action of PG, further degradation of the cell wall was affected by pectin methylesterases, pectin lyase, pectate lyase and cellulases. The disparity in enzymatic activity at different intervals may suggest their specific role for pathogenesis at proper timings.
Metabolomics Analysis on Antifungal Activities Produced by Penicillium oxalic...Agriculture Journal IJOEAR
—In-vitro antagonist tests such as disc diffusion and minimum inhibition concentration (MIC) were conducted against C. gloeosporioides. 1 H-NMR coupled with multivariate statistical analysis was carried out to identify possible compounds produced. Glucose crude extract exhibited the highest percent inhibition of radial growth (PIRG) with 75% and the lowest MIC value with 78 µg mL-1. For metabolomics, different metabolites produced were clustered according to the carbon sources used and gave a representative impression of the metabolites produced by P. oxalicum T3.3. The study has shown the potential of using a combination of 1 H-NMR spectroscopy and multivariate statistical analysis and their correlation with MIC in differentiating the effect of carbon sources used based on the identification of possible metabolites contributing to their differences. Findings from this work may potentially provide the basis for further studies on both antimicrobial activities against plant pathogen and elucidation of the metabolite compounds produced by P. oxalicum T3.3.
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Indigenized remote control interface card suitable for MAFI system CCR equipment. Compatible for IDM8000 CCR. Backplane mounted serial and TCP/Ethernet communication module for CCR remote access. IDM 8000 CCR remote control on serial and TCP protocol.
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Indigenized remote control interface card suitable for MAFI system CCR equipment. Compatible for IDM8000 CCR. Backplane mounted serial and TCP/Ethernet communication module for CCR remote access. IDM 8000 CCR remote control on serial and TCP protocol.
• Remote control: Parallel or serial interface
• Compatible with MAFI CCR system
• Copatiable with IDM8000 CCR
• Compatible with Backplane mount serial communication.
• Compatible with commercial and Defence aviation CCR system.
• Remote control system for accessing CCR and allied system over serial or TCP.
• Indigenized local Support/presence in India.
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• Compatible with MAFI CCR system.
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Production and optimization of lipase from candida rugosa using groundnut oilcake under solid state fermentation
1. IJRET: International Journal of Research in Engineering and Technology ISSN: 2319-1163
__________________________________________________________________________________________
Volume: 01 Issue: 04 | Dec-2012, Available @ http://www.ijret.org 571
PRODUCTION AND OPTIMIZATION OF LIPASE FROM CANDIDA
RUGOSA USING GROUNDNUT OILCAKE UNDER SOLID STATE
FERMENTATION
K. S. S. Rekha1
, Dr. M. V. V Chandana Lakshmi2
, Dr. V. Sri Devi3
,.
M.siddartha Kumar4
1,4
M.tech, 2,3
Associate Professor, Centre for Biotechnology, Department of Chemical Engineering, Andhra University,
Visakhapatnam-03, Andhra Pradesh, India,
kolli.rekha@gmail.com, mahantilakshmi@yahoo.com. vellurusridevi@yahoo.co.in,siddharthakumar.m@gmail.com
Abstract
The present work deals with the screening of microorganisms Candida rugosa NCIM 3467 and Penicillum citrinum NCIM 765 with
different agro residues – rice bran, wheat bran, groundnut oil cake, coconut oil cake and sesame oil cake for maximum production of
lipase. Among all the industrial residues, Groundnut oil cake supported the maximum lipase production by C.rugosa NCIM 3467. The
physical factors such as fermentation time, temperature, pH, inoculum age, inoculum level, initial moisture content played a vital role
in lipase production and further the yield was improved with the supplementation of carbon and organic nitrogen sources to the solid
medium. At 5 days of fermentation, 32 °C, pH 6, 5 day old culture, 15% inoculum level and at 60% initial moisture content, lipase
activity of 57.25 U/ml was obtained. Further the activity was raised to 63.35 U/ml by supplementing the substrate media with maltose
(5%w/w) and peptone (3%w/w).
Keywords: Candida rugosa, Pencillum citrinum, Solid state Fermentations, Lipase, Optimization and Characterization.
------------------------------------------------------------------******-------------------------------------------------------------------
1. INTRODUCTION
Lipases (triacylglycerol acylhydrolases, E.C.3.1.1.3) catalyze
the hydrolysis of triglycerides to glycerol and free fatty
acids at an oil – water interface [15],16]. Lipases can also
perform the reverse reaction of synthesis of triacylglycerols
from free fatty acids and glycerols [3], [6]. This property is
extensively used in trans- and inter-esterification reactions in
organic solvents to produce useful acylglycerols. Each
application requires unique properties with respect to
specificity, stability, temperature and pH dependence or ability
to catalyze synthetic ester reactions in organic solvents.
Interest in lipases has increased markedly in the last two
decades owing to their applications in oleochemical,
detergent, organic industries, leather industry,
environmental management, cosmetics and perfume
industry, biomedical and biosensors [9], [12], [2], and [17].
Lipases of microbial origin have gained considerable attention
in the field of biotechnology and a large number of microbial
strains have been used for the enzyme production. Lipase
production was studied by Candida rugosa in universal yeast
medium [7].Lipases are produced by several microorganisms,
namely bacteria, fungi, archea, eucarya as well by animals and
plants. Commercially useful lipases are usually obtained from
microorganisms that produce a wide variety of extracellular
lipases [2]. Microbial commercial lipases are mainly produced
from Pseudomonas [19], Mucor[1], Geothrium [31], Rhizopus
[13], P.chrysogenum [3], Bacillus subtilis [3], Aspergillus
oryzae [3], P. roqueforti [3]and Candida sp.[28],[5],[11] such
as C. rugosa [3], C. Antarctica [3] and C. parapsilosis [3].
Production of lipase can be significantly increased by the
careful selection of the nitrogen and carbon sources for
microbial growth and the optimization of the composition of
the growth medium [28],[5],[8] Most importantly, since lipase
is an inducible extracellular enzyme, by application of a
proper inducer the lipase production can be considerably
increased[21]. Histo-chemical study conformed that lipase
activity is exclusively associated to oil-bodies [20]. In recent
years, increasing attention has been paid to the conversion of
processing industry wastes by microbial lipases or their use as
biosensors [27], [31].
2. MATERIALS AND METHODS
2.1 Microorganism
C.rugosa NCIM 3467 and Pencillium citrinum NCIM 765
was purchased from the National Collection of Industrial
Microorganisms (NCIM) Pune, C.rogusa in yeast medium
(YM) and P.citrinum in potato dextrose agar medium (PDA)
maintained at 4°C and sub cultured once a month[5].
2. IJRET: International Journal of Research in Engineering and Technology ISSN: 2319-1163
__________________________________________________________________________________________
Volume: 01 Issue: 04 | Dec-2012, Available @ http://www.ijret.org 572
3.1 LIPASE PRODUCTION BY SSF
5g substrate was taken in a 250 ml Erlenmeyer flask and then
moistened with 5ml of salt solution and sterilized at 121°C for
30 min. The fermentation process was started by adding 1 ml
of spore suspension containing 107 spores/mL from a 7-day-
old culture grown on medium. The whole content was mixed
thoroughly and then incubated at 30 °C for 7 days.
3.2 SCREENING OF SUBSTRATE
Substrates such as rice bran (RB), groundnut oil cake (GOC),
sesame oil cake (SOC), coconut oil cake (COC) and wheat
bran (WB) were purchased from the local market,
Visakhapatnam are used for lipase production.
Microorganisms were grown under solid state fermentation
and conditions were optimized for the maximum production of
lipase. The production of enzyme was carried out considering
different parameters like selection of substrate, incubation
period, temperature, initial pH, inoculum age, inoculum level,
initial moisture levels and various carbon and nitrogen
additives etc.
3.3 EFFECT OF PHYSICAL PARAMETERS
3.3.1 Effect of Incubation time on lipase activity
The effect of incubation time on lipase activity was carried at
various time periods such as 24,48,72,96,120,144 and 168
hours in medium and incubated at 30°C. The fermentation
time was kept as constant for the consecutive experiments.
3.3.2 Effect of Incubation temperature on lipase
activity
For selection of optimum temperature for the production of
lipase the temperatures varying from 28-48°C were selected
and fermentation was carried for 7 days
3.3.3 Effect of pH on lipase activity
To study the effect of pH, the lipase activity was measured at
various pH ranging from 3 to 8. The pH was varied using
different buffers (citrate buffer for pH 3-6.phosphate buffer for
pH 7-8 and borate buffer for pH 9-10).
3.3.4 Effect of inoculum age on lipase activity
The effect of inoculum age on lipase production was carried
out using 1 to 8-day old day culture.
3. 3.5 Effect of inoculum level on lipase activity
The effect of inoculum level on lipase production was carried
out by fermentation of different inoculum levels ranging from
5-25% (v/w).
3.3.6 Effect of initial moisture content on lipase
activity
To investigate the influence of the initial total moisture content
(before autoclaving) of the substrate, the fermentation was
carried out under various initial moisture contents 40-90% of
substrate, which was adjusted with distilled water.
3.4 EFFECT OF CHEMICAL PARAMETERS
3.4.1 Effect of Carbon supplements on lipase activity
Substrate is supplemented with different carbon supplements
like maltose, lactose, sucrose and fructose at a final
concentration 5% (w/w).
3.4.2 Effect of Nitrogen supplements on lipase activity
Substrate is supplemented with different nitrogen supplements
such as yeast extract, beef extract, peptone and urea at a
concentration of 3% (w/w) were used.
3.5 EXTRACTION OF CRUDE ENZYME
100ml of distilled water was added to each flask and the
mixture was shaken for 30 min at room temperature to
facilitate the extraction of the enzymes from fermented
medium. At the end of the extraction the suspension was
squeezed through a double-layered muslin cloth and it was
centrifuged at 12,000 rpm for 5min.The clear supernatant
obtained was used as the extracellular enzyme.
3.6 LIPASE ASSAY (Titrimetric method)
Lipase activity was assayed by titrimetric method. In this
method, 1.25ml of olive oil and 4ml of 0.1M phosphate buffer
and the fermented filtrate is added 1ml in a 250ml flask and
immediately incubated at 37°C for 20 min at 60 rpm. Later the
reaction was inhibited by addition of chloroform: ethanol
mixture in the ratio of 2:1. The fatty acids liberated during the
incubation were titrated against 0.05N NaOH and the pink
color indicates the end point.
One unit of lipase is defined as the amount of enzyme that
liberates one micromole of fatty acid per minute under the
assay conditions [9].
4. RESULTS AND DISCUSSION
The present study deals with the production of lipase from
candida rugosa NCIM 3467 using groundnut oil cake under
solid state fermentation. The results are presented and
discussed in light and existing literatures. Process parameters
like fermentation time, temperature, pH, inoculum age,
inoculum level and initial moisture content were optimized.
The basal medium was supplemented with carbon and organic
nitrogen.
3. IJRET: International Journal of Research in Engineering and Technology ISSN: 2319-1163
__________________________________________________________________________________________
Volume: 01 Issue: 04 | Dec-2012, Available @ http://www.ijret.org 573
4.1 SUBSTRATE SCREENING
The selection of ideal agro-industrial residues for the enzyme
production in a SSF process depends upon several factors,
mainly with cost and availability of the substrate material.
Five different substrates groundnut oil cake, rice bran, wheat
bran, sesame oilcake and coconut oil cake were screened for
the maximum production of lipase. Maximum enzyme
production was obtained with groundnut oil cake.
4.2 SCREENING OF MICROORGANISMS
C.rugosa NCIM 3467 and P.citrinum NCIM 765 were
screened for production of lipase under SSF. C.rugosa NCIM
3467 produced relatively more production of lipase on
groundnut oil cake substrate with 42.50 U/ml of enzyme
activity (Chart 4.1). C.rugosa was selected to optimize the
process parameters for the enzyme production on groundnut
oil cake substrate under the SSF.
Chart 4.1 Lipase activity of two microorganisms using
different substrates
4.3 OPTIMIZATION OF PROCESS PARAMETERS
FOR LIPASE PRODUCTION
Any fermentation process is governed by a large number of
physical, chemical and biological factors. The protocol
adopted for the optimization of process parameters was to
evaluate the effect of individual parameters at a time and to
incorporate it at the standard level.
4.3.1 Effect of physical parameters
4.3.1.1 Effect of fermentation time on lipase activity
The production of lipase was studied by conducting the
fermentation using C.rugosa NCIM 3467 for different time
intervals (1, 2, 3, 4, 5, 6 and 7 days). The enzyme production
was increased with increasing time and maximum enzyme
activity 48.50 U/ml was obtained after 5 days of incubation
(Chart 4.2).
Jigita Padhiar [14] reported that lipase production has also
been influenced by the time course of the fermentation
process. The highest yield of lipase was recorded after 5 days
for Pseudomonas aeruginosa.
Chart 4.2 Effect of fermentation time on Lipase activity
4.3.1.2 Effect of temperature on lipase activity
The inoculated flasks were incubated at different temperatures
to determine the optimum fermentation temperature for lipase
production. The enzyme production was carried out by
C.rugosa NCIM 3467 at 28-48°C temperature range. The
optimum incubation temperature of the production of lipase
50.25 U/ml was found at 32°C (Chart 4.3). Enzyme
production was reduced when the incubation temperature was
increased above 32°C.
Thowlath Noora Parveen [30] reported that the lipase
production was increased at pH 5, 32°C, using Aspergillus
flavus.
4. IJRET: International Journal of Research in Engineering and Technology ISSN: 2319-1163
__________________________________________________________________________________________
Volume: 01 Issue: 04 | Dec-2012, Available @ http://www.ijret.org 574
Chart 4.3 Effect of temperature on lipase activity.
4.3.1.3 Effect of pH on lipase activity
The effect of pH can be measured by fixing the process
variables incubation period for 5 days and the temperature at
32°C, the optimum values obtained from the previous steps.
The amount of lipase produced is measured at each pH 4-9.
The maximum lipase activity of 51.25 U/ml is observed at pH
6.0 (Chart 4.4). Enzyme production was decreased when
increased above pH 6. As the metabolic activities of the
microorganisms are very sensitive to changes in pH and is
found to be affected if pH level is higher or lower compared to
the optimum value.
Falony [10] reported that the optimum lipase activity was
obtained at pH 6, by using Aspergillus niger.
Chart 4.4 Effect of pH on lipase activity.
4.3.1.4 Effect of inoculum age on lipase activity
The effect of inoculum age on lipase production was studied
by conducting the fermentation with different inoculum age.
The substrate was inoculated with 1 to 8-day old day culture.
The five day old culture gave maximum production of lipase
53.28 U/ml (Chart 4.5).
Sarada Sarkar [25] results showed that the lipase activity was
maximum in the 3 days old culture.
Chart 4.5 Effect of inoculum age on lipase activity.
5. IJRET: International Journal of Research in Engineering and Technology ISSN: 2319-1163
__________________________________________________________________________________________
Volume: 01 Issue: 04 | Dec-2012, Available @ http://www.ijret.org 575
4.3.1.5 Effect of inoculum level on lipase activity
The effect of inoculum level on lipase production was studied
by conducting the fermentation with different inoculum levels.
Various inoculum levels (5-25%) were tried to study their
effect on enzyme production. The higher enzyme production
56.25 U/ml (Chart 4.6) was obtained at 15% (v/w). High
inoculum levels are inhibitory in nature. Inoculum level as
compared to low or high inoculum levels while lower
inoculum levels require more time for fermenting the
substrates in SSF [4].
Tehreema iftikar [29] reported that the highest lipase activity
was found at 1.0 ml of inoculums size by using Rhizopus
oligosporousuv -31
Chart 4.6 Effect of inoculum level on lipase activity.
4.3.1.6 Effect of initial moisture content on lipase
activity
The optimum initial moisture content for lipase production
was determined by adjusting the initial moisture content of the
fermentation substrate to varying levels of 40-90%. A moisture
level of 60% (v/w) was found to be optimum for lipase
production 57.25 U/ml (Chart 4.7).
Saima Rehman [24] reported that the maximum lipase activity
was observed at 60% initial moisture content,using
Penicillium notatum.
Chart 4.7 Effect of initial moisture content on Lipase activity.
4.3.2 Chemical parameters
4.3.2.1 Effect of carbon supplements on lipase activity
Influence of various carbon supplements on enzyme
production was studied by adding various supplements namely
maltose, lactose, sucrose and fructose 5% (w/w) to
fermentation medium. Of the four different carbon
supplements used as enrichment, maltose was a good carbon
supplement which gave maximum lipase activity 60.25 U/ml
(Chart 4.8).
Saima Rehman [24] reported that the presence of 2% maltose
resulted in the highest lipase yield of 4626 U/gds using
Penicillium notatum. Rao [23] and Mahanta [18] reported that
the presence of maltose in the growth media enhanced the
lipase production by Pseudomonas aeruginosa and Candida
rugosa respectively.
Chart 4.8 Effect of different carbon supplements on lipase
activity.
6. IJRET: International Journal of Research in Engineering and Technology ISSN: 2319-1163
__________________________________________________________________________________________
Volume: 01 Issue: 04 | Dec-2012, Available @ http://www.ijret.org 576
4.3.2.2 Effect of nitrogen supplements on lipase
activity
Influence of organic nitrogen supplements on enzyme
production was studied by adding various supplements namely
yeast extract, peptone, beef extract and urea 3% (w/w) to
fermentation media. Of the four different nitrogen
supplements used as enrichment, it was observed peptone a
good nitrogen supplement which gave maximum lipase
production 63.35 U/ml (Chart 4.9).
Shu yang sun [26] reported that the supplementation of 2%
(w/w) of peptone gave the highest activity of 16.855 U/kg
using Rhizopus chinensis. Peptone was also found to
efficiently improve the production of extracellular lipase.
Chart 4.9 Effect of nitrogen supplements on lipase activity
SUMMARY AND CONCLUSION
Lipases have found a wide range of applications in various
industries such as chemical industry, food industry, textile,
leather and pulp industries, synthetic chemistry and detergents
etc. SSF has revealed the possibilities of effective utilization
of agro-industrial residues for value addition through
biotechnological means, lipase production by SSF using agro-
industrial residues brings down the cost of production and it
also provide an alternative path for the effective utilization of
such nutrients rich agro-industrial residues.
Fermentation time of 5 days, temperature of 32°C, pH 6.0, 5th
day old culture, 15% inoculum level and 60% initial moisture
content were found to be the optimum conditions for lipase
production and the maximum lipase activity was found to be
57.25 U/ml. Different carbon and nitrogen sources were used
as supplements at 5% (w/w) and 3% (w/w) respectively, to
determine their effect on enzyme yield. Maltose (60.25 U/ml)
and Peptone (63.35 U/ml) supplements improved enzyme
activity.
The present study indicates that groundnut oil cake is potential
substrate for lipase production using C.rugosa NCIM 3467
under SSF. Results obtained with study are comparable hence
process can be exploited for commercial application after a
comprehensive study.
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[1] Abbas H, Hiol A, Deyris V, Comeau L., Isolation and
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[2] Akimoto M, Nagashima Y and Sato D., A kinetic study
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[5] Benjamin S and Pandey., Optimization of liquid media
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[6] Bezbradica D, Mijin D, Siler-Marinkovic S, Knezevic
Z, Mol J., A kinetic study of Candida rugosa from
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[7] Ciafardini G, Zullo B.A and Iride A., Lipase production
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[8] Dalmau E, Montesinos J. L, Lotti M and Casas C.,
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[9] Freire D.M, Teles E.M.F, Bon E.P.S and Lippel San’t
Anna G., Lipase production by Penicillum restrictum in
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