Cloning, purification, and characterization of the organic solvent tolerant β-glucosidase, OaBGL84, from Olleya aquimaris DAU311

다국어 초록 (Multilingual Abstract)

A marine bacterium, Olleya aquimaris DAU311, was isolated from Goraebul beach in the Republic of Korea. This strain had β-glucosidase activity on Luria–Bertani esculin plates. The β-glucosidase, oabgl84, was isolated, cloned, and sequenced, based on fosmid library. The gene encoded novel β-glucosidase and consisted of an open reading frame of 2304 bp, which encodes 768 amino acids. The deduced amino acid sequence had 99% identity to Olleya sp. VCSM12, 84% identity to Olleya marilimosa, and 78% similarity to Lacinutrix sp. Hel_I_90. OaBGL84 belongs to the glycoside hydrolase family 3, and it was visualized using SDS-PAGE, approximately 84 kDa. The optimal temperature and pH of OaBGL84 were analyzed as 40 °C and 6.0, respectively, using pNPG as substrate. The Km and Vmax values for OaBGL84 were 1.35 mM and 25.3 μM/s, respectively. Furthermore, OaBGL84 activity was completely inhibited by Cu2+ and Hg2+ ions. OaBGL84 demonstrated extraordinary stability until 50% (v/v) benzene, n-hexane, or toluene. These results indicate that OaBGL84 is useful candidate to degrade cellulose or soy isoflavone in the organic solvents for various biotechnological applications.

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