MSA - Cell and molecular biology/Physiology & Genetics
Jain, Devendra , upadhyay, mukesh , Pareek, Shalini , Singh, Abhijeet , verma, HN .
Optimization for production of amylases from Penicillium chrysogenum MTCC 160.
Amylases are the important enzymes employed in the starch processing industries for the hydrolysis of polysaccharides. Although amylases can be obtained from various sources, but the enzymes from microbial sources, especially from fungi, generally fulfill industrial demand. Fungal amylases have successfully replaced chemical hydrolysis of starch in processing industries. Penicillium chrysogenum MTCC# 160 was optimized for the production of amylase when grown on starch containing basal salt medium exhibited the highest amylase production after 5 days of incubation. The optimum temperature and pH of amylases was recorded at 30°C and 5.0-7.5 respectively. Further studies on the activity of enzyme was done with metal ions at 20mM per ml of reaction mixture, Na+ and Ca ++ ions gave more than 90% relative activity where as metal ions Cu++,Cd+,Hg ++,Mn++,Li+ affected approximate 60% activity. Phenol showed maximum inhibition (0.00U/ml) followed by ammonium (1.5U/ml) and acetic acid (4.5U/ml) where as aniline and benzene dose not affected and showed 95% relative activity followed by ethyl alcohol and aldehyde (85%). Seven inhibitors were studied reported that β-mercaptoethanol is greatly inhibiting the activity (0.00U/ml) followed by urea (7U/ml) and sodium nitrate (8U/ml).The Km value of this enzyme was found 0.04 mg per ml with soluble starch as the substrate. Zymographical analyses indicated that there are three alleles/loci responsible for amylases and their Rf are A16.6, A216, A337.3 respectively. The activities of isozymes were also studied with ions and inhibitors. Ions like Fe++,Mg++,K+, Zn++ did not inhibited two isozymes (A16.6,A216) while Co++,Li+,Cu++,Mn++,Cd++ produced only one band (Rf A16.6. Na +and Ca++ produced all bands whereas all isozymes are inhibited by Hg++. β mercaptoethanol inhibited all isozymes while sodium bisulphate, sodium hypochlorite, hydrogen peroxide inhibited third isozyme.Only first isozyme was produced by sodium nitrite,urea,and EDTA.
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1 - Jaipur National University, Biotechnology, School of Life Sciences, Jaipur National University, Jagatpura , jaipur, rajasthan, 302025, india
Presentation Type: Oral Paper:Papers for Topics
Location: White Pine/Cliff Lodge - Level C
Date: Monday, July 27th, 2009
Time: 10:45 AM