Purification and Characterization of a Maltotetraose-Forming Alkaline (alpha)-Amylase from an Alkalophilic Bacillus Strain, GM8901.

Applied and Environmental Microbiology
T U KimY C Shin

Abstract

An alkalophilic bacterium, Bacillus sp. strain GM8901, grown at pH 10.5 and 50(deg)C, produced five alkaline amylases in culture broth. At an early stage of the bacterial growth, amylase I (Amyl I) was produced initially and then, as cultivation progressed, four alkaline amylases, Amyl II, Amyl III, Amyl IV, and Amyl V, were produced from proteolytic degradation of Amyl I. A serine protease present in the culture medium was believed to be involved in Amyl I degradation. We purified Amyl I from the culture supernatant by ammonium sulfate precipitation, heparin-Sepharose CL-6B column chromatography, phenyl-Toyopearl column chromatography, and Mono Q HR5/5 high-performance liquid chromatography. The molecular weight of Amyl I was estimated to be about 97,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Amyl I had an extremely high optimal pH of 11.0 to 12.0 and was stable in a broad pH range of 6.0 to 13.0. Amyl I had an optimal temperature of 60(deg)C and was stable up to 50(deg)C. Thermostability was increased in the presence of Ca(sup2+) and soluble starch. The enzyme required metal ions such as Ca(sup2+), Mg(sup2+), Cu(sup2+), Co(sup2+), Ag(sup+), Zn(sup2+), and Fe(sup2+) for its enzyme activity and was inhibi...Continue Reading

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Citations

Jul 16, 2008·Applied Biochemistry and Biotechnology·Raoudha Ellouz GhorbelSemia Ellouz Chaabouni
Mar 25, 2014·Journal of Industrial Microbiology & Biotechnology·Zhenghui LuYanhe Ma
Jul 20, 2006·Applied Microbiology and Biotechnology·Dilli Rani NagarajanChandraraj Krishnan
Oct 11, 2008·Journal of Industrial Microbiology & Biotechnology·Dastager G SyedAshok Pandey
May 11, 2010·Canadian Journal of Microbiology·Olha I KubrakVolodymyr I Lushchak
Jan 23, 2010·Applied Microbiology and Biotechnology·Pratibha DheeranDilip K Adhikari
Aug 19, 2014·Applied Biochemistry and Biotechnology·Francois N Niyonzima, Sunil S More
Apr 12, 2011·Journal of Industrial Microbiology & Biotechnology·Indira P SarethySanjay Gupta
May 29, 2013·Journal of Basic Microbiology·Balu Jancy Kalpana, Shunmugiah Karutha Pandian
Dec 10, 1999·Microbiology and Molecular Biology Reviews : MMBR·K Horikoshi
Mar 10, 2006·Bioresource Technology·Rajendra Kumar SaxenaPreeti Nayyar

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