open access

https://hdl.handle.net/2115/62912

A Novel Eliminase from a Marine Bacterium That Degrades Hyaluronan and Chondroitin Sulfate

英語

Marine Bacterium

2014-10-03

American Society for Biochemistry and Molecular Biology (ASBMB)

Journal of Biological Chemistry

289(40)

27886-27898

Lyases cleave glycosaminoglycans (GAGs) in an eliminative mechanism and are important tools for the structural analysis and oligosaccharide preparation of GAGs. Various GAG lyases have been identified from terrestrial but not marine organisms even though marine animals are rich in GAGs with unique structures and functions. Herein we isolated a novel GAG lyase for the first time from the marine bacterium Vibrio sp. FC509 and then recombinantly expressed and characterized it. It showed strong lyase activity toward hyaluronan (HA) and chondroitin sulfate (CS) and was designated as HA and CS lyase (HCLase). It exhibited the highest activities to both substrates at pH 8.0 and 0.5 m NaCl at 30 °C. Its activity toward HA was less sensitive to pH than its CS lyase activity. As with most other marine enzymes, HCLase is a halophilic enzyme and very stable at temperatures from 0 to 40 °C for up to 24 h, but its activity is independent of divalent metal ions. The specific activity of HCLase against HA and CS reached a markedly high level of hundreds of thousands units/mg of protein under optimum conditions. The HCLase-resistant tetrasaccharide Δ4,5HexUAα1-3GalNAc(6-O-sulfate)β1-4GlcUA(2-O-sulfate)β1-3GalNAc(6-O-sulfate) was isolated from CS-D, the structure of which indicated that HCLase could not cleave the galactosaminidic linkage bound to 2-O-sulfated d-glucuronic acid (GlcUA) in CS chains. Site-directed mutagenesis indicated that HCLase may work via a catalytic mechanism in which Tyr-His acts as the Brønsted base and acid. Thus, the identification of HCLase provides a useful tool for HA- and CS-related research and applications.

学術雑誌論文

Version of Record

This research was originally published in Journal of Biological Chemistry. Wenjun Han, Wenshuang Wang, Mei Zhao, Kazuyuki Sugahara and Fuchuan Li. A Novel Eliminase from a Marine Bacterium That Degrades Hyaluronan and Chondroitin Sulfate. Journal of Biological Chemistry. 2014; Vol:289(40) p.27886-27898 © the American Society for Biochemistry and Molecular Biology

0021-9258

1083-351X