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A high-throughput screening method for amino acid dehydrogenase

Author:
Xu, Jian-Miao, Fu, Fang-Tian, Hu, Hai-Feng, Zheng, Yu-Guo
Source:
Analytical biochemistry 2016 v.495 pp. 29-31
ISSN:
0003-2697
Subject:
abscisic acid, copper, fluorescence, ions, mutants, mutation, rapid methods, screening
Abstract:
A simple and rapid screening method for amino acid dehydrogenase (e.g., leucine dehydrogenase, LDH) has been developed. It relies on a competitive relationship between a non-fluorescent Cu(II)–calcein complex and amino acid (e.g., l-2-aminobutyric acid, l-ABA). When ABA was introduced to a Cu(II)–calcein solution, it bound with the Cu(II) ions and this released calcein from the complex, which was detected as strong fluorescence. The principle of this high-throughput screening method was validated by screening an LDH mutant library. Compared with other methods, this method provided much quicker l-ABA detection and screening for leucine dehydrogenase mutations.
Agid:
5285727