Abstract

Pyridoxal 5′-phosphate (PLP) is the coenzyme of more than 140 enzymes and is widely used in various fields. In this study, to enhance the production of PLP in Escherichia coli BL21, the recombinant strain E. coli BL21/pETDuet-1-pdxj-zwf-dxs was constructed. The concentration of PLP in this strain was 82.69 mg/L, which was increased by 1.38-fold as compared to that in E. coli BL21. Glucose, yeast extract, and pH had an obvious impact on the concentration of PLP, and their optimal levels were 34.89 g/L, 31.17 g/L, and 10.07, respectively. The concentration of PLP under the optimal condition reached 2.23 g/L. The time-course analysis showed that the highest concentration of PLP was 2.32 g/L in recombinant strain after the induction for 12 h by 0.1 mM IPTG in a 1 L shake flask, which was increased by 38.76-fold as compared to that in E. coli BL21. This study provides a good basis for the efficient production of PLP in E. coli BL21.

摘要

5'-磷酸吡哆醛 (PLP) 是 140 多种酶的辅酶，广泛应用于各个领域。本研究为提高大肠杆菌BL21中PLP的产量，构建了重组菌株大肠杆菌BL21/pETDuet-1-pdxj-zwf-dxs。该菌株中PLP的浓度为82.69 mg/L，比大肠杆菌中的浓度增加了1.38倍BL21。葡萄糖、酵母抽提物和pH对PLP浓度有明显影响，其最佳水平分别为34.89 g/L、31.17 g/L和10.07。最佳条件下PLP的浓度达到2.23 g/L。时程分析表明，重组菌株在1 L摇瓶中用0.1 mM IPTG诱导12 h后，PLP的最高浓度为2.32 g/L，比E中增加了38.76倍。 . 大肠杆菌BL21。本研究为大肠杆菌BL21中PLP的高效生产提供了良好的基础。