Purification and characterization of 2-haloacid dehalogenase from marine bacterium Paracoccus sp DEH99, isolated from marine sponge Hymeniacidon perlevis

Jinyou Zhang, Yanjuan Xin, Xu-Peng Cao, Song Xue, Wei Zhang

    Research output: Contribution to journalArticle

    13 Citations (Scopus)

    Abstract

    2-haloacid dehalogenases constitute a group of dehalogenases which are capable of dehalogenating the halogenated organic compounds. So far, the 2-haloacid dehalogenases have been found in many bacteria, but not in Paracoccus genus. In the present study, one enzyme 2-haloacid dehalogenase (designated as Deh99), induced by DL-2-chloropropionate (DL-2-CPA), was purified from the marine bacterium Paracoccus sp. DEH99, isolated from marine sponge Hymeniacidon perlevis. The enzyme of Deh99 was purified to homogeneity by ammonium sulfate precipitation, ion exchange chromatography (Q-Sepharose HP), and Superdex 200 gel filtration chromatography. The molecular weight of Deh99 was estimated to be 25.0 kDa by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE), and 50.0 kDa natively by gel filtration chromatography. The enzyme of Deh99 stereospecifically dehalogenated L-2-CPA to produce D-lactate, with an apparent Michaelis-Menten constant (K m) value of 0.21 mmol L-1 for L-2-CPA. The optimal pH and temperature for Deh99 activity were 10.0 and 40°C, respectively. The enzyme of Deh99 acted on short-carbon-chain 2-haloacids, with the highest activity towards monochloroacetate. The activity of Deh99 was slightly affected by DTT and EDTA, but strongly inhibited by Cu2+ and Zn2+. The enzyme of Deh99 shows unique substrate specificity and inhibitor sensitivities compared to previously characterized 2-haloacid dehalogenases and is the reported one about purified 2-haloacid dehalogenase isolated from the bacteria of Paracoccus genus.

    Original languageEnglish
    Pages (from-to)91-96
    Number of pages6
    JournalJournal Ocean University China
    Volume13
    Issue number1
    DOIs
    Publication statusPublished - 2014

    Keywords

    • 2-Haloacid dehalogenase
    • marine bacterium
    • marine sponge
    • Paracoccus sp.
    • purification

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