工业微生物2017，Vol.47Issue(5)：1-7,7.DOI:10.3969/j.issn.1001-6678.2017.05.001
重组大肠杆菌全细胞催化合成莱鲍迪苷D
Production of rebaudioside D by recombinant Escherichia coil whole cell catalyst
摘要
Abstract
Rebaudioside D (RD) is a minor component of stevia glycosides isolated from stevia rebaudiana with high sweetness.In this study,the whole cell biotransformation of rebaudioside A (RA) to RD was investigated.A glycosyltransferase encoding sequence eugt11 was cloned from Oryza sativa and inserted into the expression vector pETDuet-1 to yield pETDuet-eugt11.The obtained recombinant plasmid was transformed into E.coli BL21 (DE3).The recombinant enzyme was successfully expressed in E.coli BL21 (DE3) after induction and further purified by Ni + affinity chromatography.The optimal catalytic conditions were also investigated:the optimal pH of 100 mmol/L sodium phosphate buffer was 8.0 and the optimal temperature was 42 ℃.The highest production level of RD reached to 123.6 mg/L in the presence of 0.1 mmol/L ZnCl2,60 mmol/L sodium citrate,1％ (v/v) dimethylbenzene,12 mmol/L UDPG with 160 mg/L fresh cells for 24 h productivity incubation.关键词
甜菊糖/莱鲍迪苷D/EUGT11/重组大肠杆菌/全细胞催化Key words
steviol glycosides/rebaudioside D/recombinant E.coli/EUGT11/whole cell catalysis引用本文复制引用
杨玉凤,费理文,李建华,郝千里,杨靖亚,王勇..重组大肠杆菌全细胞催化合成莱鲍迪苷D[J].工业微生物,2017,47(5):1-7,7.基金项目
国家自然科学基金(31670099) （31670099）
上海市自然科学基金(14JC1406900 （）
17ZR1435000) （）
中国科学院分子植物科学卓越创新中心部署项目(CEMPS2016004). （CEMPS2016004）
