葡萄糖激酶与 ATP 再生系统偶联生产 6-磷酸葡萄糖。
Production of glucose-6-phosphate by glucokinase coupled with an ATP regeneration system.
机构信息
State Key Lab of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.
出版信息
World J Microbiol Biotechnol. 2014 Mar;30(3):1123-8. doi: 10.1007/s11274-013-1534-7. Epub 2013 Oct 29.
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-Na₂, 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 °C water bath for 1 h.
构建了一个利用乙酰磷酸（ACP）通过乙酸激酶（ACKase）进行葡萄糖-6-磷酸（G-6-P）生产与三磷酸腺苷（ATP）再生耦联的过程。从大肠杆菌 K12 中扩增和克隆了 glk 和 ack 基因，并将其分别克隆到 pET-28a(+)中，然后转化到大肠杆菌 BL21（DE3）中，将重组菌株分别命名为 pGLK 和 pACK。pGLK 中的葡激酶（glkase）和 pACK 中的 ACKase 均以可溶形式过表达。使用非常少量的 ATP 从葡萄糖和 ACP 高效生产 G-6-P。当将含有 10 mM 葡萄糖、20 mM ACP-Na₂、0.5 mM ATP、5 mM Mg²⁺、50 mM 磷酸钾缓冲液（pH 7.0）、4.856 U glkase 和 3.632 U ACKase 的反应溶液置于 37°C 水浴中 1 小时时，转化率大于 97%。
Zotero 插件