过表达果糖脱氢酶促进氧化葡萄糖酸杆菌高效合成5-酮果糖  

Efficient Production of 5-KetoFructose by Gluconobacter oxydans via Overexpression of Fructose Dehydrogenase

作　　者：钱雪 赵晨休 林金萍[1,2,3] 魏东芝[1,2,3] QIAN Xue;ZHAO Chenxiu;LIN Jinping;WEI Dongzhi(School of Biotechnology,East China University of Science and Technology,Shanghai 200237,China;State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology,Shanghai 200237,China;Key Laboratory of Biocatalysis and Intelligent Manufacturing,China National Light Industry,East China University ofScience and Technology,Shanghai 200237,China)

机构地区：[1]华东理工大学生物工程学院,上海200237 [2]华东理工大学生物反应器工程国家重点实验室,上海200237 [3]华东理工大学中国轻工业生物催化与智能制造重点实验室,上海200237

出　　处：《食品与生物技术学报》2024年第6期58-66,共9页Journal of Food Science and Biotechnology

基　　金：国家“十三五”重点研发计划项目(2021YFC2100300)。

摘　　要：5-酮果糖(5-KetoFructose,5-KF)具有与D-果糖相似的甜度和自然甜味,且不能被人体代谢而具有低热量的优点,是一种具有发展前景的非营养型天然甜味剂。为了获得5-酮果糖高产菌株,作者将来源于Gluconobacter japonicus的膜结合的果糖脱氢酶基因在Gluconobacter oxydans中过表达,通过对不同的宿主、表达载体和启动子的筛选,获得一株能够高效合成5-KF的工程菌QT-10。在3 L发酵罐中进行补料发酵优化,最终采用恒速连续流加果糖的方式,该工程菌可在82 h内消耗质量浓度为650 g/L的D-果糖,5-KF的产量高达608 g/L,产率为94.6%,时空产率为7.41 g/(L·h)。5-Ketofructose(5-KF)has a similar sweet taste quality as D-fructose,and cannot be metabolized by humans,giving it a low caloric value.These properties illustrate 5-KF can become a potential candidate for a non-nutritive natural sweetener.In this study,the 5-KF producing strain was constructed via overexpression of membrane-bound fructose dehydrogenase gene from Gluconobacter japonicus in Gluconobacter oxydans.By screening different expression hosts,expression vectors and promoters,the engineered strain QT-10 which can efficiently synthesis 5-KF was obtained.The fed fermentation process was optimized in a 3 L fermenter.Finally,650 g/L D-fructose could be completely transformed in 82 h with continuous fructose-feeding strategy,and up to 608 g/L 5-KF was reached with the yield of 94.6%and the space-time yield of 7.41 g/(L·h).

关 键 词：5-酮果糖 果糖脱氢酶 氧化葡萄糖酸杆菌 重组表达 发酵优化 

分 类 号：Q815[生物学—生物工程]