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Characterization of a GH family 8 β-1,3-1,4-glucanase with distinctive broad substrate specificity from Paenibacillus sp. X4

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An Erratum to this article was published on 12 December 2014

Abstract

A β-1,3-1,4 glucanase gene of Paenibacillus sp. X4, bglc8H, was cloned and characterized. BGlc8H was predicted to be a protein of 409 amino acid residues, including a signal peptide of 31 amino acids. The mature enzyme was predicted to have 378 amino acid residues; ITS molecular mass and pI were estimated as 41,561 Da and 7.61, respectively. BGlc8H belongs to glycoside hydrolase family 8 (GH8). Site-directed mutants of Glu95 and Asp156 of BGlc8H showed a near-complete loss of activity, indicating that they are catalytically-active residues. Unlike other GH8 members, BGlc8H had broad substrate specificity and hydrolyzed barley-β-D-glucan > chitosan > carboxymethyl-cellulose > and lichenan. BGlc8H had a lower ratio of lichenase/barley-β-d-glucanase activities compared to GH16 enzymes. BGlc8H was optimally active at pH 5 and 50 °C, except for barley-β-d-glucanase (40 °C) and chitosanase (pH 7) activities. BGlc8H hydrolyzed cello-oligosaccharides (G3–G6) to G3 and G2 but not to G1. Ca2+ increased the activity and thermostability of BGlc8H for lichenan suggesting its use for the saccharification of cellulosic biomass.

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Acknowledgments

This work was supported by the Mid-Career Researcher Program through NRF (NRF-2008-0061310), by the Cooperation Research Program through the Rural Development Administration (PJ007449201006), and partially by the Basic Research Program through NRF (NRF-2013R1A1A4A01013394), Korea.

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Authors and Affiliations

  1. Department of Agricultural Chemistry, Sunchon National University, Suncheon, 540-950, Republic of Korea

    Han Beur Na, Sung Kyum Kim, Jungho Kim & Hoon Kim

  2. Research Institute of Life Pharmaceutical Sciences, Sunchon National University, Suncheon, 540-950, Republic of Korea

    Won Kyeong Jung & Hoon Kim

  3. Department of Pharmacy, Sunchon National University, Suncheon, 540-950, Republic of Korea

    Yu Seok Jeong, Hee Jung Kim & Hoon Kim

  4. Division of Applied Life Science, Gyeongsang National University, Chinju, 660-701, Republic of Korea

    Han Dae Yun

  5. Division of Chemical & Bioengineering, Konkuk University, Seoul, 143-701, Republic of Korea

    Jung-Kul Lee

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  1. Han Beur Na
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Correspondence to Hoon Kim.

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Na, H.B., Jung, W.K., Jeong, Y.S. et al. Characterization of a GH family 8 β-1,3-1,4-glucanase with distinctive broad substrate specificity from Paenibacillus sp. X4. Biotechnol Lett 37, 643–655 (2015). https://doi.org/10.1007/s10529-014-1724-x

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