期刊
FEBS OPEN BIO
卷 5, 期 -, 页码 528-534出版社
WILEY
DOI: 10.1016/j.fob.2015.06.002
关键词
Cyclodextrin glucanotransferase; Bacillus sp.; Gamma-cyclodextrin; Random mutagenesis; DNA shuffling
资金
- European Social Fund (SAB) [080937195]
- German Research Foundation (DFG)
- Leipzig University within the program of Open Access Publishing
Cyclodextrin glucanotransferase (EC 2.4.1.19) from the alkaliphilic Bacillus sp. G-825-6 converts starch mainly to gamma-cyclodextrin (CD8). A combination of error-prone PCR and DNA shuffling was used to obtain variants of this enzyme with higher product specificity for CD8 and a broad pH activity range. The variant S54 with seven amino acid substitutions showed a 1.2-fold increase in CD8-synthesizing activity and the product ratio of CD7:CD8 was shifted to 1:7 compared to 1:3 of the wild-type enzyme. Nine amino acid substitutions of the cyclodextrin glucanotransferase were performed to generate the variant S35 active in a pH range 4.0-10.0. Compared to the wild-type enzyme which is inactive below pH 6.0, S35 retained 70% of its CD8-synthesizing activity at pH 4.0. (C) 2015 The Authors. Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.
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