Kratko priopćenje
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
Sažetak
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.
Ključne riječi
taurine; L-cysteine sulfinate decarboxylase; metagenomic library; sequential error-prone PCR
Hrčak ID:
197482
URI
Datum izdavanja:
30.3.2018.
Posjeta: 1.029 *