biostudies-literatureSong WScience and Technology Project Founded by Education Department of Jiangxi ProvinceNational Natural Science Foundation of China84https://www.ebi.ac.uk/biostudies/studies/S-EPMC9925633biostudies-literatureUnknown13(3)In this paper, the cgt gene encoding cyclodextrin glycosyltransferase (CGTase) from <i>Bacillus stearothermophilus</i> was cloned into pWB980 plasmid for extracellular expression in <i>Bacillus subtilis</i> 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 r3 BiotechHeterologous expression of cyclodextrin glycosyltransferase from <i>Bacillus stearothermophilus</i> in <i>Bacillus subtilis</i> and its application in glycosyl rutin production.PMC9925633GJJ20230531600639Li XZheng JZhang YZhang MSong WfalseHeterologous expression of cyclodextrin glycosyltransferase from <i>Bacillus stearothermophilus</i> in <i>Bacillus subtilis</i> and its application in glycosyl rutin production.In this paper, the cgt gene encoding cyclodextrin glycosyltransferase (CGTase) from <i>Bacillus stearothermophilus</i> was cloned into pWB980 plasmid for extracellular expression in <i>Bacillus subtilis</i> 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 r2023-01-01T00:00:00Z2023 Mar2026-06-02T19:25:49.88Z2025-04-04T20:40:54.342ZS-EPMC99256333679885510.1007/s13205-023-03510-5