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Molecular cloning, over-expression and enzymatic characterization of an endo-acting β-1,3-glucanase from marine bacterium Mesoflavibacter zeaxanthinifaciens S86 in Escherichia coli

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Abstract

Glucanases are involved in degradation of glucans. Here, we report a new endo-β-1,3-glucanase Mzl86 identified in Mesoflavibacter zeaxanthinifaciens S86. The deduced amino-acid sequence of Mzl86 showed highest similarity (45.1%) with Leeuwenhoekiella blandensi and thus placed in glycosyl hydrolase family 16. Purified recombinant protein (rMz186) showed an optimum enzyme activity against laminarin at 50℃ and pH 8. The enzyme was stable at 50℃ for 1 hour (maintaining 80% of its maximum activity) and was strongly activated (187%) in the presence of 2.5 mM manganese. Substrate-specific activities of rMzl86 against laminarin, barley β-glucan and lichenan were 261, 128 and 115 unit/mg, respectively. rMzl86 degraded laminarioligosaccharides (lager than biose) and laminarin while producing mainly biose and glucose. Molecular and biochemical properties reveal that rMzl86 shares typical features of β-1,3-glucanase (EC 3.2.1.39) and thus is a potential candidate for use in agriculture, drug, chemical and bioethanol industries.

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Authors and Affiliations

  1. Global Bioresources Research Center, KIOST, Ansan, 426-744, Korea

    Youngdeuk Lee, Ji-Hyun Lee, Won-Bo Shim, Su-Jin Lee, Soo-Jin Heo, Do-Hyung Kang & Chulhong Oh

  2. Department of Plant Pathology and Microbiology, Texas A&M University, 2132 TAMU, College Station, TX, 77843-2132, Texas, USA

    Won-Bo Shim

  3. Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju, 690-756, Korea

    Don Anushka Sandaruwan Elvitigala & Jehee Lee

  4. College of Veterinary Medicine, Chungnam National University, Daejeon, 305-764, Korea

    Mahanama De Zoysa

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  1. Youngdeuk Lee
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  2. Ji-Hyun Lee
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  3. Won-Bo Shim
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Correspondence to Chulhong Oh.

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Lee, Y., Lee, JH., Shim, WB. et al. Molecular cloning, over-expression and enzymatic characterization of an endo-acting β-1,3-glucanase from marine bacterium Mesoflavibacter zeaxanthinifaciens S86 in Escherichia coli . Ocean Sci. J. 49, 425–432 (2014). https://doi.org/10.1007/s12601-014-0040-7

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  • DOI: https://doi.org/10.1007/s12601-014-0040-7

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