To develop a method combining enzymatic catalysis and resting-cell biotransformation to produce allitol from low cost substrate d-glucose. The recombinant E. coli expressing d-psicose-3-epimerase (DPE), ribitol dehydrogenase (RDH) and formate dehydrogenase (FDH) for allitol production from d-fructose was constructed. The optimizations of the cell catalytic conditions and the cell cultivation conditions were made. Then, 63.4 g allitol L−1 was obtained from 100 g d-fructose L−1 in 4 h catalyzed by the recombinant E. coli cells. In order to decrease the substrate cost, d-glucose was used as the substrate instead of d-fructose and immobilized glucose isomerase was used to convert d-glucose into d-fructose. In order to simplify allitol production process from d-glucose, one-pot reaction using the mixed catalysts was used and the reaction conditions were optimized. Finally, 12.7 g allitol L−1 was obtained from 50 g d-glucose L−1 catalyzed by the mixed catalysts of immobilized glucose isomerase and the recombinant E. coli cells. Allitol can be efficiently produced from low cost substrate d-glucose by using the method combining enzymatic catalysis and resting-cell biotransformation, which is the first report.

中文翻译：

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使用固定化葡萄糖异构酶和表达 d-psicose-3-差向异构酶、核糖醇脱氢酶和甲酸脱氢酶的重组大肠杆菌优化 d-葡萄糖生产蒜醇

开发一种将酶催化和静息细胞生物转化相结合的方法，以从低成本底物 d-葡萄糖生产阿糖醇。构建了表达 d-psicose-3-差向异构酶 (DPE)、核糖醇脱氢酶 (RDH) 和甲酸脱氢酶 (FDH) 的重组大肠杆菌，用于从 d-果糖生产阿糖醇。对细胞催化条件和细胞培养条件进行了优化。然后，在重组大肠杆菌细胞的催化下，4 小时内从 100 g d-果糖 L-1 中获得 63.4 g allitol L-1。为了降低底物成本，使用d-葡萄糖代替d-果糖作为底物，并使用固定化葡萄糖异构酶将d-葡萄糖转化为d-果糖。为了简化从 d-葡萄糖生产阿糖醇的过程，采用混合催化剂的一锅法反应，优化了反应条件。最后，在固定化葡萄糖异构酶和重组大肠杆菌细胞的混合催化剂催化下，50 g d-葡萄糖L-1 得到12.7 g allitol L-1。采用酶催化和静息细胞生物转化相结合的方法，可以从低成本的底物d-葡萄糖有效地生产Allitol，这是首次报道。