文献类型：期刊文献

中文题名：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

英文题名：Efficient Production of 5-KetoFructose by Gluconobacter oxydans via Overexpression of Fructose Dehydrogenase

作者：钱雪[1];赵晨休[1];林金萍[1,2,3];魏东芝[1,2,3]

机构：[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

年份：2024

卷号：43

期号：6

起止页码：58

中文期刊名：食品与生物技术学报

外文期刊名：Journal of Food Science and Biotechnology

收录：CSTPCD;;北大核心:【北大核心2023】；CSCD:【CSCD_E2023_2024】；

基金：国家“十三五”重点研发计划项目(2021YFC2100300)。

语种：中文

中文关键词：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

外文关键词：5-KetoFructose;fructose dehydrogenase;Gluconobacter oxydans;recombinant expression;fermentation optimization

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).