黑曲霉固体发酵产木聚糖酶的响应面优化设计及其酶学性质的研究

本研究采用响应面法的中心组合对影响黑曲霉(aspergillus niger JL 15)固体发酵产木聚糖酶的条件进行了优化.以橘皮粉为基质,补充碳源、氮源,以及含水量和发酵时间对木聚糖酶产量具有显著影响(P<0.05).模拟二次多项式回归预测模型,并建立自变量与响应值的回归方程,获得各因素的最佳水平,即:甘油、硫酸铵的添加量分别为4.2%和3.1%,含水量为61%,发酵时间为73.4 h,木聚糖酶活性最大预测值达922.9 U/g干发酵产物,验证值为917.7 U/g干发酵产物,高出基础培养基酶活3.2倍.酶学性质分析研究表明,黑曲霉木聚糖酶(XylA)的最适温度和最适pH分别为55oC和pH 5.0.XylA的K_m和V_(max)值分别为9.24 mg/mL和54.05 μmol/min/mL.Mn~(2+)、Zn~(2+)和斗和Mg~(2+)对XylA活性具有促进作用,而Fe~(3+)和cu~(2+)对XylA活性具有明显抑制作用.HPLC分析结果表明,XylA的水解桦木木聚糖和麸皮不溶性木聚糖的产物均为木糖至木六糖,主要产物为木三糖.

Optimization of Solid State Fermentation Conditions for Xylanase Production by Aspergillus niger Using Response Surface Methodology and the Enzymatic Properties

The production of xylanase(XylA) bya newlyisolated Aspergillus niger JL15 strain in solidstate fermentation(SSF) on orange peel was optimized by response surface methodology(RSM).Results revealed that four factors of concentrations of added glycerin and ammonium sulfate,moisture content,fermentation time had significant effect on the XylA production(P<0.05).The maximum xylanase activity(917.7 U/g dry fermentation product) obtained at 4.2% glycerin,3.1%(NH4)2SO4 by employing orange peel as the solid substrat...