文献类型：期刊文献

英文题名：Production of glucose-6-phosphate by glucokinase coupled with an ATP regeneration system

作者：Yan, Bingkun[1];Ding, Qingbao[1];Ou, Ling[1];Zou, Zhi[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2014

卷号：30

期号：3

起止页码：1123

外文期刊名：WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY

收录：;EI(收录号：20140717331426);WOS:【SCI-EXPANDED(收录号:WOS:000331101000036)】；

语种：英文

外文关键词：Glucose-6-phosphate; ATP; Glucose kinases; Acetate kinase

摘要：A process of glucose-6-phosphate (G-6-P) production coupled with an adenosine triphosphate (ATP) regeneration system was constructed that utilized acetyl phosphate (ACP) via acetate kinase (ACKase). The genes glk and ack from Escherichia coli K12 were amplified and cloned into pET-28a(+), then transformed into E. coli BL21 (DE3) and the recombinant strains were named pGLK and pACK respectively. Glucokinase (glkase) in pGLK and ACKase in pACK were both overexpressed in soluble form. G-6-P was efficiently produced from glucose and ACP using a very small amount of ATP. The conversion yield was greater than 97 % when the reaction solution containing 10 mM glucose, 20 mM ACP-Na-2, 0.5 mM ATP, 5 mM Mg2+, 50 mM potassium phosphate buffer (pH 7.0), 4.856 U glkase and 3.632 U ACKase were put into 37 A degrees C water bath for 1 h.