Cloning and Expression of a Putative Chitinase-Encoding Gene from a Thermophilic Marine Bacterium Rhodothermaceae sp. MEBiC09517

Abstract

Rhodothermaceae sp. MEBiC09517, a thermophilic marine bacteria with considerable evolutionary distance from previously reported, was isolated from a sediment near an wood processing company in Incheon City. Based on the completed genome data, a gene(ROT0835) among glycoside hydrolases affiliated with variety of GH families was selected. The ROT0835 has 1,140 nucleotides encoding 379 amino acids, and Mmolecular weight was predicted to be 42,354Da. The amino acid sequence showed identity with chitinase from Salinibacter ruber (56%) and glycoside hydrolase from Rhodothermus marinus (54%). ROT0835 was cloned and expressed through E.coli system using T-easy vector and pET-24a vector. An expressed protein was purified by affinity chromatography & size exclusion, and optimal condition for enzyme activity was estimated.a, a gene(ROT0835) among glycoside hydrolases affiliated with variety of GH families was selected. The ROT0835 has 1,140 nucleotides encoding 379 amino acids, and Mmolecular weight was predicted to be 42,354Da. The amino acid sequence showed identity with chitinase from Salinibacter ruber (56%) and glycoside hydrolase from Rhodothermus marinus (54%). ROT0835 was cloned and expressed through E.coli system using T-easy vector and pET-24a vector. An expressed protein was purified by affinity chromatography & size exclusion, and optimal condition for enzyme activity was estimated.