Purification and characterization of a cold-adapted κ-carrageenase from Pseudoalteromonas sp. ZDY3

Dongying Zhao; Bo Jiang; Yue Zhang; Wenhui Sun; Zhongji Pu; Yongming Bao

doi:10.1016/j.pep.2020.105768

Purification and characterization of a cold-adapted κ-carrageenase from Pseudoalteromonas sp. ZDY3

Protein Expr Purif. 2021 Feb:178:105768. doi: 10.1016/j.pep.2020.105768. Epub 2020 Oct 6.

Authors

Dongying Zhao¹, Bo Jiang², Yue Zhang¹, Wenhui Sun¹, Zhongji Pu¹, Yongming Bao³

Affiliations

¹ School of Bioengineering, Dalian University of Technology, Dalian, 116024, China.
² School of Bioengineering, Dalian University of Technology, Dalian, 116024, China. Electronic address: bojiang@dlut.edu.cn.
³ School of Bioengineering, Dalian University of Technology, Dalian, 116024, China; School of Ocean Science and Technology, Dalian University of Technology, Panjin, 124221, China. Electronic address: biosci@dlut.edu.cn.

PMID: 33035660
DOI: 10.1016/j.pep.2020.105768

Abstract

κ-Carrageenase (EC3.2.1.83) is a class of glycoside hydrolase, which can be used for hydrolysis of κ-carrageenan to κ-carrageenan oligosaccharides. In this study, a bacterium, identified as Pseudoalteromonas sp. ZDY3 isolated from rotten algae, was capable to degrade κ-carrageenan. The κ-carrageenase produced by Pseudoalteromonas sp. ZDY3 was purified to homogeneity and named as CgkZDY3. The accurate molecular mass of CgkZDY3 was determined through LC-HRMS, and a posttranslational removal of C-terminal end of the protein was discovered. CgkZDY3 had strict hydrolysis specificity to κ-carrageenan, the values of K_m and k_cat/K_m of CgkZDY3 were 3.67 mg mL^-1 and 53.0 mL mg^-1 s^-1, respectively. CgkZDY3 was a cold-adapted κ-carrageenase with excellent storage stability of both the temperature below 35 °C and a wide pH range, and was an endo-type κ-carrageenase with high hydrolysis rate, oligosaccharides with different degrees of polymerization can be obtained by controlling the hydrolysis time, and the final products were κ-neocarrabiose and κ-neocarratetraose. These properties are of great significance for production of κ-carrageenan oligosaccharides with different polymerization degrees under process control.

Keywords: Carrageenan; Enzyme characterization; Pseudoalteromonas sp.; κ-Carrageenan oligosaccharides; κ-Carrageenase.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acclimatization*
Bacterial Proteins* / chemistry
Bacterial Proteins* / isolation & purification
Cold Temperature
Glycoside Hydrolases / chemistry
Glycoside Hydrolases / isolation & purification
Pseudoalteromonas / enzymology*

Substances

Bacterial Proteins
Glycoside Hydrolases