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

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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.

Keywords

β-Glucosidase Glycoside hydrolase family 3 Olleya aquimaris Organic solvent tolerance OaBGL84 

Notes

Acknowledgments

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2011-0008619), the Ministry of Education (2017-0434), and the Ministry of Science and ICT (2017-0182).

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Copyright information

© The Korean Society for Applied Biological Chemistry 2018

Authors and Affiliations

  1. 1.Department of BiotechnologyDong-A UniversityBusanRepublic of Korea

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