绿色木霉复合木聚糖酶固态发酵条件优化.docVIP

绿色木霉复合木聚糖酶固态发酵条件优化摘要：目的研究不同发酵条件对绿色木霉产复合木聚糖酶性能的影响。方法采用固体发酵培养方式，通过改变发酵条件，包括碳源、氮源的种类、浓度及接种量，测定相应条件下发酵产物的酶活性来确定发酵工艺。结果绿色木霉产酶的最佳碳源为玉米芯和麸皮（4∶6）；氮源为硫酸铵（4％）；接种量为1×107个孢子/g培养基。30 ℃固体培养70 h，发酵后用无菌水（pH 5.0）28 ℃浸提3 h测定酶活性。结论在优化的培养条件下，木聚糖酶的最高酶活性达到 267 U/g。 关键词：绿色木霉；复合木聚糖酶；固体发酵 中图分类号：Q556+.2文献标识码：A文章编号：1672-979X(2007)03-0005-03 Condition Optimization on Complex Xylanase by Trichoderma viride in Solid-state Fermentation SU Yi-shan, ZHANG Xiao-yuan, ZHU Xi-qiang*, GUO Xue-ping, LING Pei-xue (Institute of Biopharmaceuticals of Shandong Province, Jinan 250108, China) Abstract：Objective To study the effects of fermentation conditions on the production of complex xylanase by Trichoderma viride. Methods An optimized incubation condition in solid-state fermentation was achieved by determining the enzyme activity when altering culture conditions, such as types and concentrations of carbon source, nitrogen source and inoculum volume. Results The optimized condition in solid-state fermentation ofTrichoderma viride was as follows：corncob and wheat bran（mass ratio 4:6） as carbon sources，4％ (NH4)2SO4 as nitrogen source, inoculum volume of 1×107 spores/g, incubation at 30 ℃ for 70h, respectively. The enzyme extraction obtained with sterile water at 28 ℃ for 3h was used for the activity determination. Conclusion Under the optimal condition, the activity of xylanase by Trichoderma viride reaches up to 267 U/g. Key words：Trichoderma viride；complex xylanase; solid-state fermentation 复合木聚糖酶通常包括β-1,4-木聚糖酶（EC 3.2.1.8）和木糖苷酶（EC3.2.1.37）。前者又称内切木聚糖酶，它从主链内部作用于木糖苷键，将大分子木聚糖降解成低聚木糖、木二糖及少量的木糖，而后者作用于木寡糖或木二糖的末端，释放出木糖 [1-2]。用于低聚木糖生产，需尽可能获得内切木聚糖酶活性高、木糖苷酶活性低的复合木聚糖酶。该酶可以由细菌、放线菌和霉菌产生。近年研究表明，该酶在造纸工业中可部分替代含氯的化合物用于纸浆漂白，可改善纸浆性能，减少漂白工艺中化学物质的用量，大大减少环境污染[3]。复合木聚糖酶还可用作饲料添加剂，去除戊聚糖类的抗营养因子，提高饲料的利用率和能量值，可用于农业有机废弃物的深加工处理，生产高附加值的低聚木糖产品[4]。木聚糖酶作为工业酶制剂在我国尚未大规模利用。我们研究了绿色木霉木聚糖酶固体发酵的优化条件。 1材料 1.1菌种、试剂等 绿色木霉（中科院微生物所）；玉米芯，麸皮（济南当地工厂）； 桦木木聚糖、木糖、胰蛋白胨、酵母提取物均系Sigma产品；其他试剂为分析纯；Gel Plate F254薄层层析板（Merck Germa