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Characterization and its Potential Application of Two Esterases Derived from the Arctic Sediment Metagenome

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Abstract

Two esterase genes (designated as estAT1 and estAT11, respectively) were cloned by activity-based screening of a fosmid library constructed with seashore sediment sample of the Arctic. The sequence analysis of the genes revealed that these esterase genes encoded proteins of 303 and 312 amino acids, respectively, and showed 40–50% identities to members of the hormone-sensitive lipase (HSL) family retaining a catalytic triad with a conserved GDSAG sequence and an oxyanion hole (HGGG). The esterases genes were overexpressed in Escherichia coli by co-expressing GroEL-GroES chaperonine, and the recombinant proteins (rEstAT1 and rEstAT11) were purified to homogeneity. The purified EstAT1 and EstAT11 were active in a broad range of temperature from 20 to 40°C with an optimum temperature at 30°C. The activation energies of rEstAT1 and rEstAT11 to hydrolyze p-nitrophenyl esters of butyrate were determined to be 12.65 kcal/mol and 11.26 kcal/mol, respectively, indicating that they are cold-adapted esterases. The purified EstAT1 and EstAT11 could hydrolyze racemic ofloxacin esters, and further rEstAT11 hydrolyzed preferentially (S)-racemic ofloxacin butyl ester with an enantiomeric excess (eep) value of 70.3%. This work represents an example that develops enzymes from the Arctic using metagenomic approach, potentially applicable to chiral resolution of heat-labile substrates.

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Acknowledgments

This work was supported by KORDI in-house program (PE98230) and the Marine & Extreme Genome Research Center Program, Ministry of Land, Transport and Maritime Affairs, Republic of Korea. We are very grateful to the Norwegian community at Ny-Ålesund and Dr. Hong Kum Lee at Korea Polar Research Institute for the support during the Arctic field study on July 2003.

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

    Jeong Ho Jeon, Jun-Tae Kim, Sung Gyun Kang, Jung-Hyun Lee & Sang-Jin Kim

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  1. Jeong Ho Jeon
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  2. Jun-Tae Kim
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  3. Sung Gyun Kang
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  4. Jung-Hyun Lee
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  5. Sang-Jin Kim
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Correspondence to Sang-Jin Kim.

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Jeong Ho Jeon and Jun-Tae Kim equally contributed to this study.

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Jeon, J.H., Kim, JT., Kang, S.G. et al. Characterization and its Potential Application of Two Esterases Derived from the Arctic Sediment Metagenome. Mar Biotechnol 11, 307–316 (2009). https://doi.org/10.1007/s10126-008-9145-2

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