一种用于NAD⁺再生的高效NADH-铁氰化物脱氢酶与漆酶偶联物。

A high effective NADH-ferricyanide dehydrogenase coupled with laccase for NAD(+) regeneration.

作者信息

Wang Jizhong, Yang Chengli, Chen Xing, Bao Bingxin, Zhang Xuan, Li Dali, Du Xingfan, Shi Ruofu, Yang Junfang, Zhu Ronghui

机构信息

Department of Bioengineering, Nanjing University of Science & Technology, Nanjing, 210094, People's Republic of China.

Dalian Institute of Biotechnology, Liaoning Academy of Agricultural Science, Dalian, 116024, People's Republic of China.

出版信息

Biotechnol Lett. 2016 Aug;38(8):1315-20. doi: 10.1007/s10529-016-2106-3. Epub 2016 May 5.

DOI:10.1007/s10529-016-2106-3

PMID:27146212

Abstract

OBJECTIVES

To find an efficient and cheap system for NAD(+) regeneration

RESULTS

A NADH-ferricyanide dehydrogenase was obtained from an isolate of Escherichia coli. Optimal activity of the NADH dehydrogenase was at 45 °C and pH 7.5, with a K m value for NADH of 10 μM. By combining the NADH dehydrogenase, potassium ferricyanide and laccase, a bi-enzyme system for NAD(+) regeneration was established. The system is attractive in that the O2 consumed by laccase is from air and the sole byproduct of the reaction is water. During the reaction process, 10 mM NAD(+) was transformed from NADH in less than 2 h under the condition of 0.5 U NADH dehydrogenase, 0.5 U laccase, 0.1 mM potassium ferricyanide at pH 5.6, 30 °C CONCLUSION: The bi-enzyme system employed the NADH-ferricyanide dehydrogenase and laccase as catalysts, and potassium ferricyanide as redox mediator, is a promising alternative for NAD(+) regeneration.

摘要

目的

寻找一种高效且廉价的NAD(+)再生系统

结果

从大肠杆菌分离株中获得了一种NADH-铁氰化物脱氢酶。该NADH脱氢酶的最佳活性温度为45°C，pH值为7.5，NADH的K m值为10μM。通过将NADH脱氢酶、铁氰化钾和漆酶相结合，建立了一种用于NAD(+)再生的双酶系统。该系统具有吸引力，因为漆酶消耗的O2来自空气，反应的唯一副产物是水。在反应过程中，在pH 5.6、30°C条件下，0.5 U NADH脱氢酶、0.5 U漆酶、0.1 mM铁氰化钾的情况下，10 mM NAD(+)在不到2小时内从NADH转化而来。结论：以NADH-铁氰化物脱氢酶和漆酶为催化剂，铁氰化钾为氧化还原介质的双酶系统是NAD(+)再生的一种有前景的替代方案。