Abstract
κ-carrageenan oligosaccharides exhibit various biological activities. Enzymatic degradation by κ-carrageenase is safe and controllable. Therefore, κ-carrageenases have captured more and more attentions. In this study, a κ-carrageenase encoding gene, cgkX, was cloned from Pseudoalteromonas sp. QY203 with degenerate and inverse PCR. It comprised an ORF of 1194 bp in length, encoding a protein with 397 amino acid residues. CgkX is a new member of glycoside hydrolase family 16. The deduced amino acid sequence shared a high similarity with CgkX of Pseudoalteromonas κ-carrageenase; however, the recombinant CgkX showed different biochemical characteristics. The recombinant enzyme was most active at pH 7.0 and 55°C in the presence of 300 mmol L−1 NaCl. It was stable in a broad range of acidity ranging from pH 3.0 to pH 10.0 when temperature was below 40°C. More than 80% of its activity was maintained after being incubated at pH 3.6–10.0 and 4°C for 24 h. CgkX retained more than 90% of activity after being incubated at 40°C for 1 h. EDTA and SDS (1 mmol L−1) did not inhibit its activity. CgkX hydrolyzed κ-carrageenan into disaccharide and tetrasaccharide as an endo-cleaver. All these characteristics demonstrated that CgkX is applicable to both κ-carrageenan oligosaccharide production and κ-carrageenase structure-function research.
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Xu, X., Li, S., Yang, X. et al. Cloning and characterization of a new κ-carrageenase gene from marine bacterium Pseudoalteromonas sp. QY203. J. Ocean Univ. China 14, 1082–1086 (2015). https://doi.org/10.1007/s11802-015-2652-7
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DOI: https://doi.org/10.1007/s11802-015-2652-7