An Efficient and Economic Method for Detecting the Activity of NADH Dependent Enzyme

China Biotechnology

2012, Vol. 32

Issue (04): 72-75 DOI:

An Efficient and Economic Method for Detecting the Activity of NADH Dependent Enzyme

XIAO Li-xia^1,2, SHI Ming-lei², WANG Yang², ZHANG Yan², SHEN Wen-long², LI De-bin², ZHAO Yu-jun¹, ZHAO Zhi-hu²

1. Institute of Animal Husbandry and Veterinary Medicine, Shenyang Agriculture University, Shenyang 110016, China;
2. Beijing Institute of Biotechnoloy, Beijing 100071, China

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Abstract

Objective: To establish a NADH depending enzyme activity detection method. Methods: FDH and LeuDH were co-expressed in E.coli and appropriate substrate was added; the FDH activity was measured by NH₃ production and the LeuDH activity measured by TLC analysis, which were also detected by measuring the NADH absorbance. The results from both methods were compared. Results: Both NH₃ production and the NADH absorbance detection proved FDH activity; at the same time, tertiary leucine production and the NADH absorbance detection proved LeuDH activity. Conclusion: The detection of NADH absorbance changes could be employed in measuring the absorbance of NADH.

Key words： FDH NADH NAD⁺ LeuDH

Received: 09 December 2011 Published: 25 April 2012

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XIAO Li-xia, SHI Ming-lei, WANG Yang, ZHANG Yan, SHEN Wen-long, LI De-bin, ZHAO Yu-jun, ZHAO Zhi-hu. An Efficient and Economic Method for Detecting the Activity of NADH Dependent Enzyme. China Biotechnology, 2012, 32(04): 72-75.

URL:

https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2012/V32/I04/72

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