首页|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

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

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

5-KetoFructosefructose dehydrogenaseGluconobacter oxydansrecombinant expressionfermentation optimization

钱雪、赵晨休、林金萍、魏东芝

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华东理工大学生物工程学院,上海 200237

华东理工大学生物反应器工程国家重点实验室,上海 200237

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

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

国家"十三五"重点研发计划项目

2021YFC2100300

2024

食品与生物技术学报
江南大学

食品与生物技术学报

CSTPCD北大核心
影响因子:0.674
ISSN:1673-1689
年,卷(期):2024.43(6)
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