TY - JOUR
T1 - Biochemical characterization of a cold-adapted κ-carrageenase from Colwellia echini
T2 - A biotechnological tool for efficient preparation of carrageenan oligosaccharides
AU - Wei, Yanshang
AU - Jiang, Ling
AU - Yao, Zhong
AU - Zhu, Benwei
AU - Du, Yuguang
N1 - Publisher Copyright:
© 2023 Elsevier Ltd
PY - 2023/12
Y1 - 2023/12
N2 - κ-Carrageenase (EC 3.2.1.83) is a glycoside hydrolase that efficiently breaks down κ-carrageenan into κ-carrageenan oligosaccharides (κ-COS). In this study, a novel GH16 family κ-carrageenase called CeCgkB was cloned from the genome of Colwellia echini. Escherichia coli BL21 (DE3) was used to express CeCgkB. CeCgkB has a complete length of 987 bp, encoding 329 amino acid residues, and a molecular weight of 37.44 kDa. Biochemical characterization indicated that CeCgkB exhibits cold-adapted properties and was able to reach a maximum activity of 1530 U/mg at pH 8.0 and 35 °C. Kinetic parameters showed that CeCgkB has a low Km value (2 mg/mL), indicating high substrate affinity. As an endo-type κ-carrageenase, CeCgkB can degrade κ-carrageenan into products of varying DP, with a preference for DP4, DP6, and DP10. ESI-MS analysis revealed that the major products of CeCgkB consist of κ-carratetraose, κ-carrahexaose, and κ-carradecaose. As a result of CeCgkB's high activity, cold-adapted, and principal products, it is a promising biotechnological tool for the efficient preparation of κ-carrageenan oligosaccharides.
AB - κ-Carrageenase (EC 3.2.1.83) is a glycoside hydrolase that efficiently breaks down κ-carrageenan into κ-carrageenan oligosaccharides (κ-COS). In this study, a novel GH16 family κ-carrageenase called CeCgkB was cloned from the genome of Colwellia echini. Escherichia coli BL21 (DE3) was used to express CeCgkB. CeCgkB has a complete length of 987 bp, encoding 329 amino acid residues, and a molecular weight of 37.44 kDa. Biochemical characterization indicated that CeCgkB exhibits cold-adapted properties and was able to reach a maximum activity of 1530 U/mg at pH 8.0 and 35 °C. Kinetic parameters showed that CeCgkB has a low Km value (2 mg/mL), indicating high substrate affinity. As an endo-type κ-carrageenase, CeCgkB can degrade κ-carrageenan into products of varying DP, with a preference for DP4, DP6, and DP10. ESI-MS analysis revealed that the major products of CeCgkB consist of κ-carratetraose, κ-carrahexaose, and κ-carradecaose. As a result of CeCgkB's high activity, cold-adapted, and principal products, it is a promising biotechnological tool for the efficient preparation of κ-carrageenan oligosaccharides.
KW - Cold-adapted
KW - Degradation
KW - Enzyme characterization
KW - κ-Carrageenase
UR - http://www.scopus.com/inward/record.url?scp=85171764727&partnerID=8YFLogxK
U2 - 10.1016/j.fbio.2023.103170
DO - 10.1016/j.fbio.2023.103170
M3 - 文章
AN - SCOPUS:85171764727
SN - 2212-4292
VL - 56
JO - Food Bioscience
JF - Food Bioscience
M1 - 103170
ER -