通过共表达 d-阿洛酮糖 3-差向异构酶和木糖异构酶从 d-葡萄糖生产 d-阿洛酮糖。
Production of d-psicose from d-glucose by co-expression of d-psicose 3-epimerase and xylose isomerase.
机构信息
Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, CAS Key Laboratory of Renewable Energy, Guangdong Provincial Key Laboratory of New and Renewable Energy Research and Development, China; University of Chinese Academy of Sciences, China.
Guangzhou Institute of Energy Conversion, Chinese Academy of Sciences, CAS Key Laboratory of Renewable Energy, Guangdong Provincial Key Laboratory of New and Renewable Energy Research and Development, China.
出版信息
Enzyme Microb Technol. 2017 Oct;105:18-23. doi: 10.1016/j.enzmictec.2017.06.003.
d-Psicose has been drawing increasing attention in recent years because of its medical and health applications. The production of d-psicose from d-glucose requires the co-expression and synergistic action of xylose isomerase and d-psicose 3-epimerase. To co-express these genes, vector pET-28a(+)-dual containing two T7 promoters and RBS sites and an Multiple Cloning Sites was constructed using the Escherichia coli expression plasmid pET-28a(+). The xylose isomerase gene from E. coli MG1665 and the d-psicose 3-epimerase gene from Agrobacterium tumefaciens CGMCC 1.1488 were cloned and co-expressed in E. coli BL21(DE3). After 24h incubation with the dual enzyme system at 40°C, the sugar conversion ratio from d-glucose to d-psicose reached 10%. The optimal conditions were 50°C, pH 7.5 with Co and Mg. The d-psicose yields from sugarcane bagasse and microalgae hydrolysate were 1.42 and 1.69g/L, respectively.
d-阿洛酮糖近年来因其在医学和健康方面的应用而受到越来越多的关注。d-阿洛酮糖由 d-葡萄糖生产需要木糖异构酶和 d-阿洛酮糖 3-差向异构酶的共表达和协同作用。为了共表达这些基因，使用大肠杆菌表达质粒 pET-28a(+)构建了含有两个 T7 启动子和 RBS 位点以及多个克隆位点的载体 pET-28a(+)-dual。从大肠杆菌 MG1665 中克隆了木糖异构酶基因，从根癌农杆菌 CGMCC 1.1488 中克隆了 d-阿洛酮糖 3-差向异构酶基因，并在大肠杆菌 BL21(DE3)中进行了共表达。在 40°C 下用双酶系统孵育 24 小时后，d-葡萄糖转化为 d-阿洛酮糖的糖转化率达到 10%。最佳条件为 50°C、pH7.5，并用 Co 和 Mg 。从甘蔗渣和微藻水解物中获得的 d-阿洛酮糖产量分别为 1.42g/L 和 1.69g/L。
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