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Published on:21 March, 2012
RGUHS Journal of Pharmaceutical Sciences, 2012; 2(1):83-89
Research Article | doi:10.5530/rjps.2012.1.12

Purification and characterization of thermostable amylase from a strain of thermoactinomyces thalpophilus KSV 17


K. Sreenivasa Rao1, P. Ellaiah2, Karnakumar V. Biradar3
1Department of Pharmaceutics, RRKS College of Pharmacy, Bidar-585402, India
2Department of Biotechnology, College of Pharmaceutical Sciences, Visakhapatnam, Andhra University, Andra Pradesh, India
3Department of Pharmacology, RRKS College of Pharmacy, Bidar-585402, India


This research reported the Purifi cation and Characterization of Thermostable Amylase from a strain of T. Thalpophilus KSV 17. The result showed that the purifi ed enzyme specifi c activity of 145.80 U mg–1, this was an increase of 21 fold than the crude enzyme extract. The analysis of SDS- polyacrylamide gel electrophoresis showed that the molecular weight of the enzyme was 52 kDa. The Optimum pH of the purifi ed enzyme showed maximum activity at pH 7.0, but the enzyme was stable in the pH range of 5.5–7.0. The optimum temperature of the purifi ed enzyme was 85°C in absence of 10 mM CaCl2 while 90°C in presence of 10 mM CaCl2., Km and Vmax values for the purifi ed enzyme were calculated as 5.2 mg ml–1, 0.45 mg ml–1 /minute respectively. The thermal stability of the purifi ed enzyme at 80°C in absence of CaCl2. and 85°C in presence of CaCl2. The purifi ed enzyme mostly inhibited by diethyl pyrocarbonate and N-bromosuccinimide and at 5 mM conc. Ca2+, Na+ and Mg2+ showed stimulatory effect while Cu2+, Zn2+, Hg2+ and Mn2+ shown inhibitory effect. The purifi ed enzyme showed good stability and compatibility on commercial detergents and on being stored for 60 minutes at 85°C.

Key words: Purifi cation, Characterization, α-Amylase, Sephadex G-200, SDS-PAGE