重组大肠杆菌全细胞催化合成D-甘油醛

Whole Cell Catalytic Synthesis of D-glyceraldehyde by Recombinant Escherichia Coli

D-甘油醛在医药、农业化学品和天然产物合成中发挥着重要作用。通过改造天蓝色链霉菌糖醇氧化酶基因,在大肠杆菌Rosetta(DE3)菌株中进行重组表达,采用全细胞催化方法将甘油转化成D-甘油醛,与传统方法相比该方法具有绿色、安全、高效的特点。对影响蛋白表达的诱导条件进行单因素优化;并对影响全细胞催化的反应条件进行了研究。结果表明,诱导剂IPTG 0.2 mmol/L、诱导温度28℃、诱导时间6 h时,构建的大肠杆菌pET-28a(+)-aldO工程菌的相对酶活性最高;菌体光密度(OD600)为10、缓冲液pH 7.5、反应温度30℃、反应时间60 min时,甘油的转化率可以达到46%,为D-甘油醛的生物合成奠定了基础。

D-glyceraldehyde plays an important role in the synthesis of pharmaceuticals,agrochemicals,and natural products.By transforming the alditol oxidase gene of Streptomyces coelicolor,the recombinant expression was carried out in E.coli Rosetta(DE3)strain,and glycerol was converted into D-glyceraldehyde by whole-cell catalysis.Compared with the traditional method,this method is green,safe and efficient.The induction conditions affecting protein expression were optimized by single factor.The reaction conditions affecting whole cell catalysis were also studied.The results showed that the relative enzyme activity of the engineered E.coli pET-28a(+)-aldO strain was the highest at an inducer IPTG concentration of 0.2 mmol/L,induction temperature of 28℃,and induction time of 6 h.When the bacterial optical density(OD600)is 10,the buffer pH is 7.5,the reaction temperature is 30℃,and the reaction time is 60 minutes,the conversion rate of glycerol can reach 46%,laying the foundation for the biosynthesis of D-glyceraldehyde.