The effect of various fermentation conditions on amylase production through shakeflask culture was investigated. Production and stabilization of amylases from aspergillus. Also ssf provides a medium that resembles the natural habitat of fungal species, unlike smf which is. Detailed description of the development and use of fermenters for the largescale cultivation of microorganisms for enzyme production is outside the scope of this volume but mention of media use is appropriate because this has a bearing on the cost of the enzyme and because media components often find their way into. The synthetic media used for the production of amylases have been a bit costlier and thats why. The present context was investigated to optimize amylase production and cell. The fermentation medium was prepared by varying ph values5. High fermentation medium cost is one of the major concerns in amylase production from. Bergey dh, holt gj 1994 bergeys manual of determinative bacteriology 9thedition. Yeast extract alone or in conjunction with other nitrogen sources such as bactopeptone and ammonium sulfate has been used for the production of. Effect of the medium composition on formation of amylase by bacillus sp. The optimal combination of the major constituents of media for. Biotechnological processes in microbial amylase production. The reaction mixture containing 1 ml of1% wv soluble starch in citrate phosphate buffer ph 6.
The present study deals with the isolation and characterisation of a moderately thermophilic bacillus and describes the effects of medium composition on the production of amylase. Amylase production by btk on jp supplemented lb medium maximum amylase activity was 460. The enzyme has been isolated from barley and rice plants. Pdf medium optimization for the production of amylase by bacillus. Pdf amylase production, purification and characterization from.
Amylase activity in starchdegrading soil isolates introduction this week you will continue our theme of industrial microbiologist by characterizing how differing reaction conditions affect enzyme reaction rates of the amylase produced by your starch degrading isolates from last week. Various physico and chemical parameters such as ph, temperature, incubation time were determined. Culture medium for amylase production by toxigenic fungi scielo. Industrial sectors with topdown and bottomup approaches are currently focusing on improving microbial amylase production levels by implementing bioengineering technologies. Pdf medium optimization for the production of amylase by. Dglucanglucanohydrolase are enzymes that are widely studied and used in industries. Brazilian archives of biology and technology 1 50 figure 1 growth and. Effect of ph on amylase production the maximum values of amylase activity mea sured during growth in the yeast nitrogen base medium, containing various carbon sources and buffered at different ph values, are listed in table 1. Production of amylase from banana peels with bacillus. Effect of medium composition on amylase production by some. Amylase production was maximum on 3rd day of incubation.
The above medium composition were dissolved in ml of distilled water after which 100ml of the medium was meas. Agar plate a consisted of 2% bactotryptone, 1% bactoyeast extract, 1% nacl and 2% agar at ph 7. Amylase is a group of enzymes whose catalytic function is to hydrolyze breakdown sugar and starch to give diverse products including dextrins, and progressively smaller polymers composed of glucose units. Bcc 02150 per bergeys manual of bacterial determination.
The optimum volume of the fermentation medium was found to be 20 ml in 100. Production methods there are mainly two methods which are used for production of. Production and partial characterization of extracellular. Adharsh rajasekar, rahulsribalaji gandhiraj dhamodharan. Amylase can be isolated from plants, animals or microorganisms. The production of extracellular amylase activity by a number of recently described amyloytic yeast species, viz.
Starch is composed of amylose and amylopectin amylose is a long unbranched chain of dglucose bonded by. The spectrophotometric stop reaction determination a 540, light path 1 cm is based on the following reaction unit definition. Five different sources were used for the isolation of the yeasts. Optimization of fermentation parameters for amylase production, the effect on. Amylase is an important and indispensable enzyme that plays a pivotal role in the field of biotechnology. Production of protease and amylase from bacillus subtilis. To identify the suitable carbon sources ability to induce amylase production in the production for amylase production by thebacillus cereus. Medium optimization for the production of amylase by bacillus subtilis rm16 in shakeflask fermentation pak. Preparation of medium used for amylase production by b. Investigation of this protein seemed of particular in terest since current work has shown that b. Cow dung is an ideal fermentation medium for amylase production. Fems microbiology letters 25 1984 169173 169 published by elsevier fem 01928 effect of medium composition on amylase product ion by some starchdegrading yeasts amylolytic enzymes. Bacillus sp according to the bergeys manual of determinative bacteriology sneath, 1986.
Isolation and characterization of amylase producing yeasts. The aamylase enzyme of this strain showed excellent stability at high temperatures and over a wide ph range. The activity of the enzyme increased between 2 and 5 gl yeast extract. Amylases have been reported to be produced by plant, animal and microbial sources, although the microbial amylase production has been reported to be most effective. Optimization of thermoalkali stable amylase production. The amino acid composition of aamylase from bacillus. The dna was isolated using promegakit according to the manufacturers instructions. Pdf studies on the alpha amylase synthesis was carried out with a moderately thermophilic. Effect of the medium composition on formation of amylase.
Among the physical parameters, the temperature and ph of the medium play an important role in. Optimization of thermoalkali stable amylase production and. Factors affecting alfa amylase production on submerged. Production and optimization of amylases using aspergillus. To obtain maximum yield of an enzyme, development of a suitable medium and culture conditions is obligatory 8,9. Characterization and optimization of amylase production in.
Amylase production by btk on pp supplemented lb medium in this medium composition, maximum. Amylase production using bacillus cereus isolated from a. Starch agar m107d can be used for the detection of starch hydrolyzing microorganisms from samples such as foods 2 and clinical samples 3. Amylase production through submerged and solid state fermentation. Bacterial strains previously isolated, identified, and stored in our laboratory were screened for amylase production in starch agar medium.
Effect of ph on the production of amylase by bacillus subtilis rm16 in shaking incubator using 1% starch as a substrate, inoculum size of 5% and temperature 40. It is produced mainly from microbial sources and is used in many industries. The effects on amylase secretion of ph, different carbon sources glucose, maltose, dextrin, soluble. Amylase production by penicillium fellutanum isolated. Amylase production is greatly reduced when glutamate or citrate is used and the amylase production is maximum when starch is used as a carbon source 19, 20, 21. The aamylase family comprises a group of enzymes with a variety of different specificities that all act on one type of substrate being.
Generally, the influence of temperature on amylase production is related to the growth of the organism. Comparative study on production of aamylase from bacillus. Amylaseproduction and process, technology, applications. Production, purifi cation, and characterization of. Production of protease and amylase from bacillus s ubtilis. The result of amylase production indicated that the composition and concentration of media greatly affected the growth and production of extracellular amylase in bacteria.
However, studies reveal that ssf is the most appropriate process in developing countries due to the advantages it offers which make it a cost effective production process. Abstract aspergillus niger ncim 820 was grown in rice bran since its a good carbon and starch source which has the ability to produce amylase. The production of extracellular amylase was highly affected by the initial ph of the medium table 1. Production medium the composition of production medium used was soluble starch 50 g, yeast extract 0. Production and medium optimization of amylase by bacillus. The above medium composition were dissolved in ml of distilled water after which 100ml of the medium was measured. The medium was prepared by weighing the following medium composition in grams per litre. Amylases are one of the most important enzymes in presentday biotechnology.
Amylase activity was assayed by measuring the reducing sugar formed by the enzymatic hydrolysis of soluble starch. Production, purification, and characterization of thermostable. It has been found that cassava mash waste water is a source of. For more information, log on to this industrial production lecture explains the amylase production process step by step in. Bacillus licheniformis tcrdc11 isolated from soil was used for aamylase production. The standard bergeys manual of systemic bacteriology was used for the. Preparation of medium used for amylase production by.
The present study was concerned with the production and partial characterization of extracellular amylase from bacillus amyloliquefaciens p001. After incubation, enzyme assay carried out at regular interval. Production and optimization of amylases using aspergillus niger. Bergeys manual of systematic bacteriology, second edition. The amylase activity was determined by dinitrosalicylic acid method. Verachtert laboratory of industrial microbiology and biochemistry, unwersity of leuven, 3030 heverlee leuven, belgium received. Media optimization, bacillus subtilis, amylase, starch. The low cost medium produced 5 times more enzyme than the high cost synthetic medium using yeast extract and peptone in shake flasks. This procedure may be used for the determination of. The present study was undertaken to isolate, identify and characterize the bacillus subtilis in the culture medium followed by alpha amylase extraction from the fermented carrier, banana peel and bacillus subtilis. Alphaamylase production by bacillus subtilis cm3 in solid. In the present study, we obtained a high amylaseproducing strain of bacteria, wanglb, and identified it as a member of the bacillus genus based on 16s rdna analysis. Effect of the medium composition on formation of amylase by.
This study aimed to prepare a readily available medium for industrial amylase production. Primary screening of extracellular enzyme amylase producing microbes amylases are starch hydrolysing enzymes. Amylase production on submerged fermentation by bacillus spp. Pdf effect of the medium composition on formation of amylase by. Amylase production was detected after flooding the plates with iodine solution hols et al. The production of the enzyme was maximum at 10 h after inoculation. The effects on amylase secretion of ph, different carbon sources glucose, maltose, dextrin, soluble starch.
Production, purification and characterization of amylase. Pour 10 ml of sterile distilled water on the slant containing fungal spores 2. Hence, the optimum temperature depends on whether the culture is mesophilic, thermophilic or psycrophilic. The production of extracellular amylase by bacillus spp was optimized in a submerged fermentation. In the recent past, there has been extensive research on. Beef extract in the medium provides nitrogen,carbon sources, amino acids and vitamins to the growing microorganisms. The fermentation medium was prepared by varying the ph values 5. Response surface methodology rsm was used to evaluate the effect of the main variables, i. In this study, the effect of various nitrogen sources in the fermentation medium on bla production was investigated.
316 118 1567 632 925 1190 126 404 1304 919 1509 1520 336 195 999 1289 1166 546 568 285 1060 1110 234 893 320 237 1468 850 1014