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A new esterase showing similarity to putative dienelactone hydrolase from a strict marine bacterium, Vibrio sp. GMD509

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Abstract

Vibrio sp. GMD509, a marine bacterium isolated from eggs of the sea hare, exhibited lipolytic activity on tributyrin (TBN) plate, and the gene representing lipolytic activity was cloned. As a result, an open reading frame (ORF) consisting of 1,017 bp (338 aa) was found, and the deduced amino acid sequence of the ORF showed low similarity (<20%) to α/β hydrolases such as dienelactone hydrolases and esterase/lipase with G–X1–S–X2–G sequence conserved. Phylogenetic analysis suggested that the protein belonged to a new family of esterase/lipase together with various hypothetical proteins. The enzyme was overexpressed in Escherichia coli and purified to homogeneity. The purified enzyme (Vlip509) showed the best hydrolyzing activity toward p-nitrophenyl butyrate (C4) among various p-nitrophenyl esters (C2 to C18), and optimal activity of Vlip509 occurred at 30°C and pH 8.5, respectively. Kinetic parameters toward p-nitrophenyl butyrate were determined as Km (307 μM), kcat (5.72 s−1), and kcat/Km (18.61 s−1 mM−1). Furthermore, Vlip509 preferentially hydrolyzed the S-enantiomer of racemic ofloxacin ester. Despite its sequence homology to dienelactone hydrolase, Vlip509 showed no dienelactone hydrolase activity. This study represents the identification of a novel lipolytic enzyme from marine environment.

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Acknowledgment

We thank Mr. H. S. Na in Enzychem for the helpful discussion. 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.

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Authors and Affiliations

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

    Sang-Yi Park, Jun-Tae Kim, Sung Gyun Kang, Jung-Hee Woo, Jung-Hyun Lee & Sang-Jin Kim

  2. Division of Biochemistry, Kangwon National University, Chuncheon, 200-701, South Korea

    Sang-Yi Park & Hyoung-Tae Choi

  3. Korea Ocean Research and Development Institute, P.O. Box 29, Ansan, 425-600, South Korea

    Sang-Jin Kim

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  1. Sang-Yi Park
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  2. Jun-Tae Kim
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Correspondence to Sang-Jin Kim.

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Park, SY., Kim, JT., Kang, S.G. et al. A new esterase showing similarity to putative dienelactone hydrolase from a strict marine bacterium, Vibrio sp. GMD509. Appl Microbiol Biotechnol 77, 107–115 (2007). https://doi.org/10.1007/s00253-007-1134-2

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  • DOI: https://doi.org/10.1007/s00253-007-1134-2

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