Abstract
Objectives
To acquire a thermostable xylanase, that is suitable for xylooligosaccharide production from pretreated corncobs, the metagenomic method was used to obtain the gene from an uncultured environmental microorganism.
Results
A thermostable xylanase-encoding gene (xyn10CD18) was cloned directly from the metagenomic DNA of cow dung compost. When xyn10CD18 was expressed in Bacillus megaterium MS941, extracellular xylansae activity at 106 IU/ml was achieved. The purified recombinant Xyn10CD18 was optimally active at pH 7 and 75 °C as measured over 10 min. It retained over 55 % of its initial activity at 70 °C and pH 7 after 24 h. Its action on birchwood xylan for 18 h liberated xylooligosaccharides with 2°–4° of polymerization, with xylobiose and xylotetraose as the main products. When pretreated corncobs were hydrolyzed by Xyn10CD18 for 18 h, the xylooligosaccharides (DP 2–4) products increased to 80 % and the xylose was just increased by 3 %.
Conclusion
Xyn10CD18 is a thermostable endoxylanase and is a promising candidate for biomass conversion and xylooligosaccharide production.
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Acknowledgments
Financial supports from the National Basic Research Program of China (2011CB710802), the National High-Tech Research and Development Plan (2011AA100905-4), the Program for Changjiang Scholars and Innovative Research Team in University (IRT1166), the National Natural Science Fund (21076159), the China postdoctoral science fund (Grant 2013M540076) and the Tianjin Science and Technology Support Plan (12ZCZDNC01700) were gratefully acknowledged.
Supporting information
Supplementary Table 1—The specific primers used for mTAIL-PCR reaction of xylanase gene.
Supplementary Fig. 1—Chromatograms of HPLC analysis of acidic electrolyzed water pretreated corncob (AEWP-corncob) hydrolysates by Xyn10CD18. The AEWP-corncob was hydrolysed by the purified Xyn10CD18 (5000 IU/g dry corncob) in 250 ml reaction system at 75 °C and pH 3-5 for 3-24 h. The Chromatograms of HPLC of the hydrolysates withdrawed at 0 h (a), 3 h (b), 12 h (c) and 24h (d) were supplied, respectively.
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Ming-zhe Sun and Hong-chen Zheng have contributed equally to this work.
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Sun, Mz., Zheng, Hc., Meng, Lc. et al. Direct cloning, expression of a thermostable xylanase gene from the metagenomic DNA of cow dung compost and enzymatic production of xylooligosaccharides from corncob. Biotechnol Lett 37, 1877–1886 (2015). https://doi.org/10.1007/s10529-015-1857-6
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DOI: https://doi.org/10.1007/s10529-015-1857-6