Fine-Tuning of Carbon Flux and Artificial Promoters in Bacillus subtilis Enables High-Level Biosynthesis of D-Allulose
文献类型： 外文期刊
作者： Zhang, Wei 1 ; Wei, Meijing 1 ; Sun, Xiaoxuan 1 ; Lu, Fuping 1 ; Guan, Lijun 3 ; Mao, Shuhong 1 ; Qin, Hui-Min 1 ;
作者机构： 1.Minist Educ, Key Lab Ind Fermentat Microbiol, Tianjin 300457, Peoples R China
2.Tianjin Univ Sci & Technol, Coll Biotechnol, Tianjin Key Lab Ind Microbiol, Natl Engn Lab Ind Enzymes, Tianjin 300457, Peoples R China
3.Heilongjiang Acad Agr Sci, Inst Food Proc, Harbin 150086, Peoples R China
关键词： D-allulose; D-allulose 3-epimerase; carbon flux; dual promoter combination; expression element optimization
期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：5.895； 五年影响因子：6.048 ）
ISSN： 0021-8561
年卷期：
页码：
收录情况： SCI
摘要： D-Allulose is an attractive rare sugar that can be used as a low-calorie sweetener with significant health benefits. To meet the increasing market demands, it is necessary to develop an efficient and extensive microbial fermentation platform for the synthesis of D-allulose. Here, we applied a comprehensive systematic engineering strategy in Bacillus subtilis WB600 by introducing D- allulose 3-epimerase (DAEase), combined with the deactivation of fruA, levDEFG, and gmuE, to balance the metabolic network for the efficient production of D-allulose. This resulting strain initially produced 3.24 g/L of D-allulose with a yield of 0.93 g of D- allulose/g D-fructose. We further screened and obtained a suitable dual promoter combination and performed fine-tuning of its spacer region. After 64 h of fed-batch fermentation, the optimized engineered B. subtilis produced D-allulose at titers of 74.2 g/L with a yield of 0.93 g/g and a conversion rate of 27.6%. This D-allulose production strain is a promising platform for the industrial production of rare sugar.
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