Abstract
To develop an enantioselective epoxide hydrolase (EHase) from marine microorganisms, marine samples were collected from a variety of marine environments. Strains isolated by the capability of living on styrene oxide (SO) were screened for retaining enantioselective EHase activities toward SO by combining spectrophotometric, GC, and HPLC analysis. Consequently, one strain, JCS358, was selected, and the sequence analysis of 16S rRNA gene showed that the strain belonged to Erythrobacter cluster. Twelve additional Erythrobacter strains from this study or acquired from culture collections were thereby tested for displaying EHase activities, and most of tested strains showed enantioselective hydrolysis toward SO and glycidyl phenyl ether. Kinetic resolution of racemic SO using whole cell of Erythrobacter sp. JCS358 was performed. Enantiopure (S)-SO could be obtained with an enantiomeric excess (ee) higher than 99% after 15 h incubation. The determination of 1-phenyl-1,2-ethanediol configuration derived from racemic SO confirmed the enantioselective hydrolyzing activity of Erythrobacter sp. JCS358.
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Acknowledgement
The authors thank the Hyper-Dolphin operation team and the R/V Natsushima team at Japan Agency for Marine-Earth Science and Technology (JAMSTEC) for sample collection in Kagoshima Bay. This work was supported by KORDI in-house program (PE97803) and the Marine & Extreme Genome Research Center Program, Ministry of Marine Affairs & Fisheries, Republic of Korea.
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Three different assays were used to determine the enantioselective hydrolysis of epoxides
1) spectrophotometric diol assay – nonstereoselective assay
2) GC analysis – measuring remaining epoxides with enantioselective resolution
3) HPLC diol analysis – measuring produced diols with enantioselective resolution
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Hwang, YO., Kang, S.G., Woo, JH. et al. Screening Enantioselective Epoxide Hydrolase Activities from Marine Microorganisms: Detection of Activities in Erythrobacter spp.. Mar Biotechnol 10, 366–373 (2008). https://doi.org/10.1007/s10126-007-9070-9
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DOI: https://doi.org/10.1007/s10126-007-9070-9