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Cloning and characterization of three epoxide hydrolases from a marine bacterium, Erythrobacter litoralis HTCC2594

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Abstract

Previously, we reported that ten strains belonging to Erythrobacter showed epoxide hydrolase (EHase) activities toward various epoxide substrates. Three genes encoding putative EHases were identified by analyzing open reading frames of Erythrobacter litoralis HTCC2594. Despite low similarities to reported EHases, the phylogenetic analysis of the three genes showed that eeh1 was similar to microsomal EHase, while eeh2 and eeh3 could be grouped with soluble EHases. The three EHase genes were cloned, and the recombinant proteins (rEEH1, rEEH2, and rEEH3) were purified. The functionality of purified proteins was proved by hydrolytic activities toward styrene oxide. EEH1 preferentially hydrolyzed (R)-styrene oxide, whereas EEH3 preferred to hydrolyze (S)-styrene oxide, representing enantioselective hydrolysis of styrene oxide. On the other hand, EEH2 could hydrolyze (R)- and (S)-styrene oxide at an equal rate. The optimal pH and temperature for the EHases occurred largely at neutral pHs and 40–55 °C. The substrate selectivity of rEEH1, rEEH2, and rEEH3 toward various epoxide substrates were also investigated. This is the first representation that a strict marine microorganism possessed three EHases with different enantioselectivity toward styrene oxide.

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Acknowledgement

This work was supported by KORDI in-house program (PE97803) and the Marine and Extreme Genome Research Center Program, Ministry of Marine Affairs and Fisheries, Republic of Korea. We would like to appreciate the Gordon and Betty Moore Foundation for the great effort on Microbial Genome Sequencing Project (Moore 155).

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Authors and Affiliations

  1. Marine Biotechnology Research Centre, Korea Ocean Research and Development Institute, Ansan, P.O. Box 29, 425-600, South Korea

    Jung-Hee Woo, Young-Ok Hwang, Sung Gyun Kang, Hyun Sook Lee & Sang-Jin Kim

  2. Division of Biology and Ocean Sciences, Inha University, Incheon, 402-751, South Korea

    Jang-Cheon Cho

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  1. Jung-Hee Woo
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Correspondence to Sang-Jin Kim.

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Woo, JH., Hwang, YO., Kang, S.G. et al. Cloning and characterization of three epoxide hydrolases from a marine bacterium, Erythrobacter litoralis HTCC2594. Appl Microbiol Biotechnol 76, 365–375 (2007). https://doi.org/10.1007/s00253-007-1011-z

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