Characterisation of a thermoalkali-stable cyclodextrin glycosyltransferase from the anaerobic thermoalkaliphilic bacterium Anaerobranca gottschalkii

Archives of Microbiology
Volker ThiemannG Antranikian

Abstract

The thermoalkaliphilic anaerobic bacterium Anaerobranca gottschalkii produces an extracellular CGTase when grown on starch at 55 degrees C and pH 9.0. The gene encoding this CGTase was cloned and successfully expressed in Escherichia coli. It encodes a protein consisting of 721 amino acids with a signal sequence of 34 amino acids. On SDS-polyacrylamide gels, the purified CGTase from A. gottschalkii displayed the expected molecular mass of 78 kDa. The recombinant enzyme was purified with a yield of 13.5% and displayed a specific activity of 210 units/mg. This CGTase, which represents the first report of a CGTase from an anaerobic thermoalkaliphile, was active at a broad range of temperature and pH, namely 55-70 degrees C and pH 5-10. It completely converted amylose, amylopectin and native starch to cyclodextrins, preferentially alpha-cyclodextrin. With a longer incubation period, the alpha-cyclodextrin to beta-cyclodextrin ratio declined. Variations in substrate type and concentration influenced the product pattern. Increasing the substrate concentration (0.5-20.0%) and glucans containing branching points (alpha-1,6 glycosidic linkages) shifted the product pattern to: beta-cyclodextin > alpha-cyclodextrin > gamma-cyclodextrin. I...Continue Reading

Citations

Apr 16, 2009·Applied Microbiology and Biotechnology·Ronan M KellyHans Leemhuis
Sep 19, 2009·Applied Microbiology and Biotechnology·Hans LeemhuisLubbert Dijkhuizen
Apr 12, 2011·Journal of Industrial Microbiology & Biotechnology·Indira P SarethySanjay Gupta
Jun 17, 2008·Applied Biochemistry and Biotechnology·Ivan PishtiyskiBoriana Zhekova
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Aug 9, 2011·Bioscience, Biotechnology, and Biochemistry·Momoko KobayashiAtsuo Kimura
Sep 6, 2012·International Journal of Molecular Sciences·Abdelnasser S S IbrahimGarabed Antranikian
May 12, 2011·Protein Expression and Purification·Sédami GnidehouJean-Louis Frendo
Dec 15, 2018·Journal of Basic Microbiology·Dhwani UpadhyayNiha M Kulshreshtha

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