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https://doi.org/10.17113/ftb.56.01.18.5415

Molecular Characterization and Directed Evolution of a Metagenome-Derived l-Cysteine Sulfinate Decarboxylase

Jie Deng ; Guangxi Key Laboratory of Mangrove Conservation and Utilization, Guangxi Mangrove Research Center, Guangxi Academy of Sciences
Qiaofen Wu ; State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Life Science and Technology
Hua Gao ; State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Life Science and Technology
Qian Ou ; State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Life Science and Technology
Bo Wu ; State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Life Science and Technology
Bing Yan ; Guangxi Key Laboratory of Mangrove Conservation and Utilization, Guangxi Mangrove Research Center, Guangxi Academy of Sciences
Chengjian Jiang ; Guangxi Key Laboratory of Mangrove Conservation and Utilization, Guangxi Mangrove Research Center, Guangxi Academy of Sciences

Abstract

L-Cysteine sulfinate decarboxylase (CSD, EC 4.1.1.29), the rate-limiting enzyme in taurine synthesis pathway, catalyzes L-cysteine sulfinic acid to form hypotaurine. Identification of the novel CSD that could improve the biosynthetic efficiency of taurine is important. An unexplored decarboxylase gene named undec1A was identified in a previous work through sequence-based screening of uncultured soil microorganisms. Random mutagenesis through sequential error-prone polymerase chain reaction was used in Undec1A. A mutant Undec1A-1180, which was obtained from mutagenesis library, had 5.62-fold higher specific activity than Undec1A at 35 °C and pH=7.0. Molecular docking results indicated that amino acid residues Ala235, Val237, Asp239, Ile267, Ala268, and Lys298 in the Undec1A-1180 protein helped recognize and catalyze the substrate molecules of L-cysteine sulfinic acid. These results could serve as a basis for elucidating the characteristics of the Undec1A-1180. Directed evolution technology is a convenient way to improve the biotechnological applications of metagenome-derived genes.

Keywords

taurine; L-cysteine sulfinate decarboxylase; metagenomic library; sequential error-prone PCR

Hrčak ID:

197482

URI

https://hrcak.srce.hr/197482

Publication date:

30.3.2018.

Article data in other languages: croatian

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