期刊
BIOCATALYSIS AND BIOTRANSFORMATION
卷 21, 期 4-5, 页码 199-207出版社
TAYLOR & FRANCIS LTD
DOI: 10.1080/10242420310001618528
关键词
glucan branching enzyme; starch; Aquifex aeolicus; thermostability; glycoside hydrolase; alpha-amylase superfamily
Glucan branching enzymes are responsible for the synthesis of alpha(1-->6) glycosidic bonds in glycogen and amylopectin. The glucan branching enzyme of the hyperthermophile Aquifex aeolicus is the most thermoactive and thermostable glucan branching enzyme described. The gene encoding this glucan branching enzyme was overexpressed in E. coli and purified using gamma-cyclodextrin affinity chromatography. Subsequently, the enzyme was subjected to a biochemical characterization. The optimum temperature for activity was 80degreesC, and the enzyme was stable up to 90degreesC. Its thermostability may be explained by the relatively high number of aromatic amino acid residues present, in combination with a relatively low number of glutamine/asparagine residues. The Km for amylose was 4 muM and the Vmax was 4.9 U/mg of protein (at optimal pH and temperature). The side-chain distribution of the branched glucan formed from amylose was determined.
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