利用重组嗜热栖热菌尿苷 - 胞苷激酶生产尿苷酸

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  • 1(天津科技大学 生物工程学院,天津,30457) 2(莲花健康产业集团股份有限公司,博士后科研工作站,河南 项城,4620 )
范晓光,博士,讲师,研究方向为代谢控制发酵。E-mail:xiaoguangfan@tust.edu.cn

网络出版日期: 2018-02-02

基金资助

天津市教委科研计划项目(2017KJ06);中国博士后科学基金(2016M601269)

Production of uridine monophosphate by recombinant uridine/cytidine kinase from Thermus thermophilus

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  • 1 (College of Biotechnology, Tianjin University of Science & Technology, Tianjin, 300457, People's Republic of China) 2 (Post-Doctoral Research Center, Lotus Health Group Company, Xiangcheng, Henan Province, 466200, People's Republic of China)

Online published: 2018-02-02

摘要

尿苷-胞苷激酶(Uridine/cytidine kinase, UCK)作为生物体核苷酸代谢补偿途径中的重要催化剂,可以催化尿苷和胞苷磷酸化成为尿苷酸(5`-uridine monophosphate, 5`-UMP)和胞苷酸(5`-cytidine monophosphate, 5`-CMP)。利用大肠杆菌Escherichia coli BL21(DE3)分别成功克隆表达了来源于嗜热栖热菌(Thermus thermophilus HB8)以及枯草芽孢杆菌(Bacillus subtilis 168)的尿苷-胞苷激酶。对比分析了不同重组尿苷-胞苷激酶的催化特性发现,来源于嗜热栖热菌的尿苷-胞苷激酶能以鸟苷三磷酸(Guanosine 5`-triphosphate, GTP)或者腺苷三磷酸(Adenosine 5`-triphosphate, ATP)作为磷酸供体催化尿苷生成尿苷酸,而来源于枯草芽孢杆菌的尿苷-胞苷激酶只有以GTP作为磷酸供体时的催化效果较好。在优化的反应条件下,使用来源于嗜热栖热菌的尿苷-胞苷激酶,6 h可催化10 mM的尿苷和10 mM的ATP生成8.74 mM尿苷酸。研究结果表明,以ATP作为磷酸供体的尿苷-胞苷激酶可作为一种新型的催化剂用于尿苷酸的研制与生产。

本文引用格式

范晓光, 贾子樊, 李国梁, 等 . 利用重组嗜热栖热菌尿苷 - 胞苷激酶生产尿苷酸[J]. 食品与发酵工业, 2018 , 44(1) : 7 . DOI: 10.13995/j.cnki.11-1802/ts.015644

Abstract

Uridine/cytidine kinase (UCK), an important catalyst in the nucleotides replenishment pathway, which is responsible for the conversion of uridine and cytidine to 5'-uridine monophosphate (UMP) and 5'-cytidine monophosphate (CMP). UCK from Thermus thermophilus HB8 and Bacillus subtilis 168 was successfully cloned and expressed in Escherichia coli BL21 (DE3) respectively. The catalytic properties of different recombinant UCK were compared. Guanosine 5`-triphosphate (GTP) and adenosine 5`-triphosphate (ATP) was the main phosphate donor of UCK from Thermus thermophilus HB8, while GTP was the main phosphate donor of UCK from Bacillus subtilis 168. Under the optimized reaction conditions, 8.74 mM UMP was obtained from 10 mM uridine and 10 mM ATP in 6 h by the UCK from Thermus thermophilus HB8. The results showed that, the ATP-dependent UCK can be used in the research and production of UMP as a new catalyst.
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