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SCREENING OF POTENTIAL ISOLATE FOR THE PRODUCTION OF AMYLASE WITH DIFFERENT AGRO WASTES AS SUBSTRATE
Abstract
Amylolytic bacteria were isolated from decayed pomegranate. The bacteria were screened for amylase activity using starch agar plate assay. Five different substrates were used namely banana peel, Gossypium oil cake, groundnut oil cake, cassava, and wheat bran; for microbial production of amylase by Submerged fermentation (SmF) and it was found that maximum amylase activity (2.005 U/ml) specific activity (0.520U/mg) was produced by bacterial isolate 8 using cassava as a substrate.
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Amylases are enzymes which hydrolyze starch molecules to give diverse products including dextrin and progressively smaller polymers composed of glucose units. These enzymes are of great significance in present day biotechnology with applications ranging from food, fermentation, textile to paper industries. Although amylases can be derived from several sources, including plants, animals and microorganisms, microbial enzymes generally meet industrial demands. In the present study, the bacteria was isolated from the soil samples. The bacteria isolated was identified as Bacillus sp. based on Staining techniques, motility test, plating on selective media and biochemical tests. Amylase production by Bacillus sp. was detected by the disappearance of blue colour in the starch agar medium around the microbial colonies after incubation. Cassava was used as the substrates for the amylase production. Solid state fermentation was carried out for the production of amylase using Bacillus sp. The effect of different carbon and nitrogen source, Temperature and pH was determined on enzyme production by Bacillus sp. Amylase activity was determined by four methods such as DNSA method, Dextrinizing activity method, decrease in starch-iodine color intensity and Plate assay.
Biotechnology as "microbial technology" is one of the most applied technologies. This technology employing microbiological systems and their products, can be explored for the utilization of various wastes and by-products originating from agriculture, food and related industries. Biotechnology has established various applications In environment, agriculture, food, dairy and meat Industries, Fermentation technology such as solid-state cultivation process can be applied in more effective way in the biotransformation and biological upgrading of crop, agricultural and food industry by-products for improved or upgraded qualities. This article discusses the potential utilisation of various wastes for pollution control, safe disposal and recycling through biotechnology. story': Genistein and Saponins.
Enzymes are among the most important products, obtained for human needs through microbial sources. A large number of industrial processes in the areas of industrial, environmental and food biotechnology utilize enzymes at some stage or the other. Current developments in biotechnology are yielding new applications for enzymes. Solid state fermentation (SSF) holds tremendous potential for the production of enzymes. It can be of special interest in those processes where the crude fermented products may be used directly as enzyme sources. This review focuses on the production of various industrial enzymes by SSF processes. Following a brief discussion of the microorganisms and the substrates used in SSF systems, and aspects of the design of fermenter and the factors affecting production of enzymes, an illustrative survey is presented on various individual groups of enzymes such as cellulolytic, pectinolytic, ligninolytic, amylolytic and lipolytic enzymes, etc.
Production of amylase and acid protease by solid state fermentation employing Aspergillus niger strain S 1 4, a mangrove isolate was evaluated. Wheat bran containing 60% moisture was used for fermentation with two inoculum sizes of 2x10 6 and 40x10 6 spores 10 g substrate -1 . The maximum amylase activity of 48.13 Ugdfs -1 was recorded after an incubation period of 4 days at temperature 30 0 C and pH 4.8 for the inoculum size 40x10 6 spores 10 g substrate -1 . The concentration of reducing sugar showed a significant negative correlation (-0.496, p<0.05) with amylase production. The peak acid protease activity was observed on day 6 (54.89 Ugdfs -1) for 2x10 6 spores 10g substrate -1 and on day 4 for 40x10 6 spores 10g substrate -1 under standard assay conditions of 30 0 C and pH 2.7. The concentration of protein in the substrate was significantly (R=0.778, p<0.05) correlated with the acid protease production.
Lignocellulosic residues obtained from crops cultivation form useful sources to be used as substrate for bioconversion processes. Sugarcane bagasse, which is a complex substrate obtained from the processing of sugar cane, is an important biomass among such sources. Due to its abundant availability, it can serve as an ideal substrate for microbial processes for the production of value added products. This paper reviews recent developments on biological processes developed on production of various products in solid state fermentation using sugarcane bagasse as the substrate and describes production of protein enriched feed, enzymes, amino acid, organic acids and compounds of pharmaceutical importance, etc. through microbial means.
Solid state fermentation was carried out using various agro- industrial wastes with the best amylase producing strain isolated from soil. Different physicochemical conditions were varied for maximum enzyme production. The strain produced about 5400 units/g of amylase at 1:3 moisture content, 20% inoculum, after 72 h of incubation with Mustard Oil seed cake as the substrate. The optimum temperature and pH of the enzyme activity were found to be 50°C and 6 respectively. The enzyme was found to be thermostable at 70°C for about 2 h without any salt. It showed stability at pH range 5-7. The metal ions as Na(+), Ca(++), Mg(++) and Co(++) enhanced the enzyme activity.
Amylases are one of the main enzymes used in industry. Such enzymes hydrolyze the starch molecules into polymers composed of glucose units. Amylases have potential application in a wide number of industrial processes such as food, fermentation and pharmaceutical industries. α-Amylases can be obtained from plants, animals and microorganisms. However, enzymes from fungal and bacterial sources have dominated applications in industrial sectors. The production of α-amylase is essential for conversion of starches into oligosaccharides. Starch is an important constituent of the human diet and is a major storage product of many economically important crops such as wheat, rice, maize, tapioca, and potato. Starch-converting enzymes are used in the production of maltodextrin, modified starches, or glucose and fructose syrups. A large number of microbial α-amylases has applications in different industrial sectors such as food, textile, paper and detergent industries. The production of α-amylases has generally been carried out using submerged fermentation, but solid state fermentation systems appear as a promising technology. The properties of each α-amylase such as thermostability, pH profile, pH stability, and Ca-independency are important in the development of fermentation process. This review focuses on the production of bacterial and fungal α-amylases, their distribution, structural-functional aspects, physical and chemical parameters, and the use of these enzymes in industrial applications.
The consumption of bi-substrates (groundnut shell and cassava waste) in order to produce an amylase enzyme by Bacillus sp. using solid stated fermentation was carried out by applying Box-Behnken design (BBD). Initial pH, incubation time, concentrations of groundnut shell (GNS) and cassava waste (CW) variables were generated for each fermentation conditions of the Bacillus sp. to optimize the amylase production. Maximum yield of amylase (866 U/mL) was achieved by Bacillus sp. at the favorable conditions of the fermentation; 60 h, pH 7.0, 2 g/L GNS and 2 g/L CW. It was revealed that the combination of bi-substrates might be used in industries to produce high amount of value added amylase.
