A Novel κ-Carrageenase from Marine Bacterium Rhodopirellula sallentina SM41: Heterologous Expression, Biochemical Characterization and Salt-Tolerance Mechanism Investigation.

Autor:	Zhang YH; College of Food and Biological Engineering, Jimei University, Xiamen 361021, China.; Fujian Provincial Engineering Technology Research Center of Marine Functional Food, Xiamen 361021, China.; Xiamen Key Laboratory of Marine Functional Food, Xiamen 361021, China., Chen YY; College of Food and Biological Engineering, Jimei University, Xiamen 361021, China.; Fujian Provincial Engineering Technology Research Center of Marine Functional Food, Xiamen 361021, China.; Xiamen Key Laboratory of Marine Functional Food, Xiamen 361021, China., Zhuang XY; College of Food and Biological Engineering, Jimei University, Xiamen 361021, China.; Fujian Provincial Engineering Technology Research Center of Marine Functional Food, Xiamen 361021, China.; Xiamen Key Laboratory of Marine Functional Food, Xiamen 361021, China., Xiao Q; College of Food and Biological Engineering, Jimei University, Xiamen 361021, China.; Fujian Provincial Engineering Technology Research Center of Marine Functional Food, Xiamen 361021, China.; Xiamen Key Laboratory of Marine Functional Food, Xiamen 361021, China., Chen J; College of Food and Biological Engineering, Jimei University, Xiamen 361021, China.; Fujian Provincial Engineering Technology Research Center of Marine Functional Food, Xiamen 361021, China.; Xiamen Key Laboratory of Marine Functional Food, Xiamen 361021, China., Chen FQ; College of Food and Biological Engineering, Jimei University, Xiamen 361021, China.; Fujian Provincial Engineering Technology Research Center of Marine Functional Food, Xiamen 361021, China.; Xiamen Key Laboratory of Marine Functional Food, Xiamen 361021, China., Yang QM; College of Food and Biological Engineering, Jimei University, Xiamen 361021, China.; Fujian Provincial Engineering Technology Research Center of Marine Functional Food, Xiamen 361021, China.; Xiamen Key Laboratory of Marine Functional Food, Xiamen 361021, China., Weng HF; College of Food and Biological Engineering, Jimei University, Xiamen 361021, China.; Fujian Provincial Engineering Technology Research Center of Marine Functional Food, Xiamen 361021, China.; Xiamen Key Laboratory of Marine Functional Food, Xiamen 361021, China., Fang BS; College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361021, China., Xiao AF; College of Food and Biological Engineering, Jimei University, Xiamen 361021, China.; Fujian Provincial Engineering Technology Research Center of Marine Functional Food, Xiamen 361021, China.; Xiamen Key Laboratory of Marine Functional Food, Xiamen 361021, China.
Jazyk:	angličtina
Zdroj:	Marine drugs [Mar Drugs] 2022 Dec 16; Vol. 20 (12). Date of Electronic Publication: 2022 Dec 16.
DOI:	10.3390/md20120783
Abstrakt:	κ-carrageenases are members of the glycoside hydrolase family 16 (GH16) that hydrolyze sulfated galactans in red algae, known as κ-carrageenans. In this study, a novel κ-carrageenase gene from the marine bacterium Rhodopirellula sallentina SM41 (RsCgk) was discovered via the genome mining approach. There are currently no reports on κ-carrageenase from the Rhodopirellula genus, and RsCgk shares a low identity (less than 65%) with κ- carrageenase from other genera. The RsCgk was heterologously overexpressed in Escherichia coli BL21 and characterized for its enzymatic properties. RsCgk exhibited maximum activity at pH 7.0 and 40 °C, and 50% of its initial activity was retained after incubating at 30 °C for 2 h. More than 70% of its activity was maintained after incubation at pH 6.0-8.0 and 4 °C for 24 h. As a marine derived enzyme, RsCgk showed excellent salt tolerance, retaining full activity in 1.2 M NaCl, and the addition of NaCl greatly enhanced its thermal stability. Mass spectrometry analysis of the RsCgk hydrolysis products revealed that the enzyme had high degradation specificity and mainly produced κ-carrageenan disaccharide. Comparative molecular dynamics simulations revealed that the conformational changes of tunnel-forming loops under salt environments may cause the deactivation or stabilization of RsCgk. Our results demonstrated that RsCgk could be utilized as a potential tool enzyme for efficient production of κ-carrageenan oligosaccharides under high salt conditions. Competing Interests: The authors declare no competing financial interest.
Databáze:	MEDLINE
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