重组大肠杆菌生产核苷磷酸转移酶的发酵条件优化  

Optimization of fermentation conditions for the production of phosphotransferase by Escherichia coli

作　　者：沈爱萍[1] 何宝龙[1] 孙丽慧[1] 郑裕国[1] 

机构地区：[1]浙江工业大学生物工程研究所,浙江杭州310014

出　　处：《发酵科技通讯》2013年第4期1-4,共4页Bulletin of Fermentation Science and Technology

基　　金：国家自然科学基金青年项目(31101351)

摘　　要：核苷磷酸转移酶能够特异地将无机焦磷酸(PPi)的磷酸根转移到核苷5′-位的羟基上,该过程并不需要ATP的参与,且具有选择性高和反应条件温和等特点。以实验室前期构建的重组大肠杆菌E.coli BL21(DE3)/pET28b-AP/PT为菌株,进行了5 L发酵罐中的培养条件优化,确定了以乳糖作为诱导剂时,最佳诱导时机OD600为7左右,诱导剂乳糖的浓度为10 g/L;确定了最适通气量为1.5 L/min。在该条件下,核苷磷酸转移酶酶活达221.4 U/L,OD600为20.4。Phosphotransferase, possessed high regiospecificity of the nucleoside phosphorylating activity to the C-5'-positon, is of high potential for the production of inosine-5′-monophosphate due to mild reaction conditions. In this paper, recombinant E. coli BL21(DE3)/pET28b-AP/PT which has expressed acid phosphatases was used, and the effects of culture conditions on cell growth and enzyme activity were investigated in 5L fermenter. The optimal conditions of culture were as following: induction time was at OD600=7, the lactose concentration was of 10 g/L, ventilation volume was of 1.5 L/min. Under the culture conditions, the enzyme activity and OD600could reach 221.4 U/L and 20.4, respectively.

关 键 词：重组大肠杆菌 核苷磷酸转移酶 发酵 条件优化 

分 类 号：Q78[生物学—分子生物学] R44[医药卫生—诊断学]