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Expression and Characterization of the Dictyoglomus thermophilum Rt46B.1 Xylanase Gene (xynB) in Bacillus subtilis

文献类型： 外文期刊

作者： Zhang, Wei 1 ; Lou, Kai 2 ; Li, Guan 1 ;

作者机构： 1.Xinjiang Univ, Coll Life Sci & Technol, Urumqi 830046, Peoples R China

2.Xinjiang Acad Agr Sci, Inst Microbe, Urumqi 830000, Peoples R China

关键词： Thermostable xylanase;Purification;Characterization;Bacillus subtilis

期刊名称：APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY （ 影响因子：2.926； 五年影响因子：2.685 ）

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收录情况： SCI

摘要： To obtain extracellular and high-level expression of the Dictyoglomus thermophilum Rt46B.1 xylanase B gene, this gene was integrated into the α-amylase gene site of a host strain of Bacillus subtilis WB800. The extreme thermophile xylanase gene was successfully integrated and expressed in the host, measured at 24±0.4 XUs/mL in the Luria broth medium supernatant. The recombinant enzyme was purified by ammonium sulfate precipitation, anion exchange chromatography, and gel filtration. The molecular mass and pI value of xylanase were estimated to be 24 kDa and 4.3, respectively. The optimal pH level and temperature of the purified enzyme were 6.5 and 85 °C, respectively. Xylanase showed reasonable activity at temperatures up to 95 °C and remained stable at 4 °C for 1 week. The purified enzyme retained most of its activity in 1 mM ethylenediaminetetraacetic acid or dithiothreitol and 0.1% Tween-20 or Triton X-100. However, strong inhibition was observed in the presence of 5 mM Mn2+, 0.5% sodium dodecyl sulfate, Tween-20, or Triton X-100; a strong stimulating effect was also observed in the presence of Fe2+. The Km and Vmax values of the recombinant xylanase for birchwood xylan were calculated to be 2.417±0.36 mg/mL and 325±41 μmol/min mg, respectively. Xylanase was found to be useful in the prebleaching process of paper pulps.

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