利用大肠杆菌工程菌生产2,3-二磷酸甘油酸的研究

Study on production of 2,3-BPG with recombinant E.coli strain

通过基因工程的方法,将表达人源BPGM(bisphosphoglycerate mutase二磷酸甘油酸变位酶,E.C.2.7.5.4.)的质粒转化入大肠杆菌内,并进行培养条件的优化摸索,通过向培养基内加入葡萄糖使大肠杆菌大量发酵生产2,3-BPG(2,3-bisphosphoglycerate,2,3-二磷酸甘油酸).结果表明,当培养基中葡萄糖质量浓度为10 g/L,诱导时间为4 h,大肠杆菌工程菌表达的2,3-BPG含量最高.如果诱导后加入终质量分数为0.5%的Tween 80可以有效促进2,3-BPG分泌到培养基中.诱导后4 h添加新培养基,补加葡萄糖可以使2,3-BPG的产量提高1倍,达到7.5 mmol/L.

2,3-BPG（2,3-bisphosphoglycerate）is one of the intermediate products in glycolytic pathway,which catalyzes by an enzyme named BPGM（bisphosphoglycerate mutase E.C.2.7.5.4）.The BPGM only exists in erythrocyte.In this paper,used recombinant E.coli strain to express BPGM in the cell,with input of glucose into the LB to induce the production of 2,3-BPG.Used malachite green method for determination of inorganic phosphate to detect 2,3-BPG.The results showed that,when the concentration of glucose is 10 g/L,and the induce time is 4 hours,it reached the highest productivity of 2,3-BPG,approximately at a level of 3.75 mmol/L.In the mean while,with the input Tween 80 to the final concentration at 0.5% can promote the 2,3-BPG to come out of the E.coli into the LB.