Research paperCloning, expression and characterization of a ι-carrageenase from marine bacterium Wenyingzhuangia fucanilytica: A biocatalyst for producing ι-carrageenan oligosaccharides

- A novel GH82 family ι-carrageenase (Cgi82A) was cloned and expressed in E. coli.
- Cgi82A possesses a low optimal reaction temperature.
- Cgi82A shows relatively high substrate-binding affinity and catalysis efficiency.
- Cgi82A can be utilized as a biocatalyst for producing ι-carrageenan oligosaccharides.

ι-Carrageenan is an algal polysaccharide widely applied in the food industry. Specific glycoside hydrolases are valuable tools for modifying polysaccharides and producing oligosaccharides. In this study, the gene of a novel GH82 family ι-carrageenase was cloned from the genome of marine bacterium Wenyingzhuangia fucanilytica. The ι-carrageenase designated as Cgi82A was expressed in Escherichia coli, and its biochemical properties, kinetic constants and hydrolysis pattern were characterized. The enzyme could reach its highest activity at 25 °C, which is lower than all hitherto reported GH82 ι-carrageenases. It was an endo-acting hydrolase, and could be utilized as a potential biocatalyst for producing ι-carrageenan oligosaccharides with different polymerization degrees. Cgi82A possessed relatively high substrate-binding affinity and catalysis efficiency indicated by its kinetic constants (Km, 1.12 μM; Kcat, 560.75 s−1). Its major end product was ι-carrageenan tetrasaccharide. The acquiring of this novel enzyme would facilitate the future application of ι-carrageenan and its oligosaccharides.

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