Heterologous expression of cyclodextrin glycosyltransferase from Bacillus stearothermophilus in Bacillus subtilis and its application in glycosyl rutin production.

ABSTRACT: In this paper, the cgt gene encoding cyclodextrin glycosyltransferase (CGTase) from Bacillus stearothermophilus was cloned into pWB980 plasmid for extracellular expression in Bacillus subtilis SCK6. Through adding a six-histidine affinity tag fused to the C-terminus, the recombinant CGTase could be purified by nickel ion affinity chromatography, and its molecular weight was approximately 76 kDa on SDS-PAGE. Then, the enzymatic properties were determined, and results were as follows: the optimum temperature and pH were identified as 40 ℃ and pH 5.0, respectively. CGTase had good tolerance to metal ions of Mn²⁺, Ca²⁺, and Mg²⁺. The enzyme activity was activated by Na⁺, Al³⁺, Fe³⁺, and Ni⁺, and it was r

SUBMITTER: Song W 

PROVIDER: S-EPMC9925633 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature