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- Title
Heterologous expression of cyclodextrin glycosyltransferase from Bacillus stearothermophilus in Bacillus subtilis and its application in glycosyl rutin production.
- Authors
Song, Wen; Zhang, Mengjie; Li, Xiaojun; Zhang, Yinjun; Zheng, Jianyong
- 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 remarkably inhibited by Cu2+ and Zn2+. To improve the aqueous solubility of rutin, CGTase was used to catalyze the transglycosylation reaction, and the conversion rate could reach as high as 80.13% under optimal conditions. Furthermore, the reaction mixture was treated with glucoamylase and microporous adsorbent resin. The yield of glycosyl-rutin was 56.1%, and its purity was 74.3%, which further improved the value of the product.
- Subjects
GEOBACILLUS stearothermophilus; GENE expression; CYCLODEXTRINS; BACILLUS subtilis; RUTIN; AFFINITY chromatography; BIOSURFACTANTS; POLYACRYLAMIDE gel electrophoresis
- Publication
3 Biotech, 2023, p1
- ISSN
2190-572X
- Publication type
Article
- DOI
10.1007/s13205-023-03510-5