Heterologous Expression and Characterization of a Glycoside Hydrolase Family 45 endo-β-1,4-Glucanase from a Symbiotic Protist of the Lower Termite, Reticulitermes speratus

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作者
Masato Otagiri
Crisanto M. Lopez
Katsuhiko Kitamoto
Manabu Arioka
Toshiaki Kudo
Shigeharu Moriya
机构
[1] Biomass Research Platform Team,Laboratory of Environmental Molecular Biology, Graduate School of Nanobioscience
[2] Biomass Engineering Program,Department of Biotechnology
[3] Research Cluster for Innovation,Graduate school of Fisheries Science and Environmental Studies
[4] RIKEN,Antibiotics Laboratory
[5] Yokohama City University,Department of Infection Biology, Leibniz Institute for Natural Product Research and Infection Biology (Hans Knöll Institute)
[6] The University of Tokyo,undefined
[7] Nagasaki University,undefined
[8] RIKEN Advanced Science Institute,undefined
[9] Friedrich-Schiller-Universität Jena,undefined
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关键词
Cellulase; Biomass; Termite; Protist;
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摘要
The termite symbiotic system is one of the efficient lignocellulose degradation systems. We tried to express and characterize a novel cellulolytic enzyme from this system. Here, we report the isolation of an endo-β-1,4-glucanase gene homolog of glycoside hydrolase family 45 from a symbiotic protistan community of Reticulitermes speratus. Heterologous expression of this gene was performed using the expression system of Aspergillus oryzae. Analysis of enzymatic properties revealed 786 μmol/min/mg protein in specific activity, a Vmax of 833.0 units/mg protein, and a Km value of 2.58 mg/ml with carboxymethyl cellulose as the substrate. Thin-layer chromatography analysis showed that RsSymEG2 produces cellobiose from cellodextrins larger than cellohexaose. This enzyme showed high specific activity like other endo-β-1,4-glucanases from the symbiotic system of termites. It means that the termite symbiotic system is a good resource for highly active endo-β-1,4-glucanases.
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页码:1910 / 1918
页数:8
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