Molecular cloning and characterization of AlgL17, a new exo-oligoalginate lyase from Microbulbifer sp. ALW1

Zedong Jiang; Yuxi Guo; Xinxia Wang; Hebin Li; Hui Ni; Lijun Li; Anfeng Xiao; Yanbing Zhu

doi:10.1016/j.pep.2019.03.015

Molecular cloning and characterization of AlgL17, a new exo-oligoalginate lyase from Microbulbifer sp. ALW1

Protein Expr Purif. 2019 Sep:161:17-27. doi: 10.1016/j.pep.2019.03.015. Epub 2019 Mar 27.

Authors

Zedong Jiang¹, Yuxi Guo², Xinxia Wang², Hebin Li³, Hui Ni¹, Lijun Li¹, Anfeng Xiao¹, Yanbing Zhu⁴

Affiliations

¹ College of Food and Biological Engineering, Jimei University, Xiamen, 361021, China; Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen, 361021, China; Research Center of Food Biotechnology of Xiamen City, Xiamen, 361021, China; Key Laboratory of Systemic Utilization and In-depth Processing of Economic Seaweed, Xiamen Southern Ocean Technology Center of China, Xiamen, 361021, China.
² College of Food and Biological Engineering, Jimei University, Xiamen, 361021, China.
³ Xiamen Medical College, Xiamen, 361008, China.
⁴ College of Food and Biological Engineering, Jimei University, Xiamen, 361021, China; Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen, 361021, China; Research Center of Food Biotechnology of Xiamen City, Xiamen, 361021, China; Key Laboratory of Systemic Utilization and In-depth Processing of Economic Seaweed, Xiamen Southern Ocean Technology Center of China, Xiamen, 361021, China. Electronic address: yanbingzhu@jmu.edu.cn.

PMID: 30926463
DOI: 10.1016/j.pep.2019.03.015

Abstract

A new alginate lyase gene of algl17 was cloned from an alginate-degrading marine bacterium Microbulbifer sp. ALW1. The gene contained 2220 bp and encoded a 739-amino acid protein classified into the PL-17 family. The recombinant alginate lyase AlgL17 was overexpressed and purified from Escherichia coli BL21 (DE3) with a molecular mass of 84.9 kDa. This enzyme showed activities towards sodium alginate, polyM and alginate oligosaccharide, but very low activity towards polyG. These results indicate that AlgL17 is a polyM-specific oligoalginate lyase. When sodium alginate was used as a substrate, the optimum reaction temperature and pH for the enzyme were 35 °C and pH 8.0, respectively. Recombinant AlgL17 was stable at 25 °C, but not stable at 30 °C and 35 °C. It showed good stability over a pH range of 5.0-8.0. The enzyme activity was increased to 1.7 times by adding NaCl to a final concentration of 0.7 M. The ability of the recombinant AlgL17 producing 4-deoxy-L-erythro-5-hexoseulose uronic acid (DEH) from sodium alginate and polyM block indicates that AlgL17 is an exo-type alginate lyase.

Keywords: Biochemical characterization; Cloning; Exo-oligoalginate lyase; Microbulbifer.

Publication types

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

MeSH terms

Alginates / metabolism
Bacterial Proteins / chemistry*
Bacterial Proteins / genetics*
Bacterial Proteins / metabolism
Cloning, Molecular*
Enzyme Stability
Escherichia coli / genetics
Escherichia coli / metabolism
Gammaproteobacteria / enzymology*
Gammaproteobacteria / genetics
Hydrogen-Ion Concentration
Kinetics
Molecular Weight
Polysaccharide-Lyases / chemistry*
Polysaccharide-Lyases / genetics*
Polysaccharide-Lyases / metabolism
Substrate Specificity
Temperature

Substances

Alginates
Bacterial Proteins
Polysaccharide-Lyases
poly(beta-D-mannuronate) lyase