产木聚糖酶芽胞杆菌的筛选及产酶条件优化.docVIP

产木聚糖酶芽胞杆菌的筛选及产酶条件优化 刘程程1,2，刘波1*(，蓝江林2，王凯3 （1.福建农林大学生命科学院，福州 350003；2福建省农业科学院农业生物资源研究所，福州 350003；3.福州大学生物工程学院，福州350002） 摘要：利用木聚糖酶选择培养基，从120株标准芽孢杆菌中筛选出37株能够形成明显透明圈的芽孢杆菌。经产酶发酵液培养基复筛，得到一株木聚糖酶活性较高的菌株FJAT-14260（沙福芽胞杆菌 B. safensis）。采用单因素实验设计和正交实验设计进行优化，在培养基优化中对产酶影响最大的因素是底物浓度，；在发酵条件优化中对产酶影响最大的因素是培养温度。优化得到FJAT-14260的最佳产酶发酵条件为：碳源(酵母粉)7 g/L,氮源(蛋白胨)9 g/L,底物（木聚糖）浓度为10 g/L，培养温度30℃，初始PH值为8，装液量为30mL/250mL.在此条件下FJAT-14260在发酵32 h时酶活力达到113585.78（U/mL)，比优化前提高了4.62倍。 关键词：芽胞杆菌 木聚糖酶 发酵条件 Screning of a xylanase producing Bacillus sp and the optical condition for xylanase production Abstract：Selective medium for xylanase was applied to screening the xylanase producing Bacillus strains and 37 of 120 type strains of Genus Bacillus were found to form transparent?circle on the plate. After secondary screening with the enzyme fermentation medium Bacillus safensis FJAT-14260 was found to be the best strain for xylanase production. Single factor experiment and orthogonal experiment were used to optimize the xylanase production conditions.The results showed that concentration of inducer was the most important factor wich affect enzyme producing in process of the medium components optimization，and cultural temperature was the most important factor in the process of fermentation conditions optimization. The best enzyme production medium after optimization was consisted of nitrogen source (yeast extract)7g/L, nitrogen source(peptone)9g/L,inducer(xylan)10g/L，then with the best fermentation temperature 30℃，pH8.0 and liquid volume 30mL/250mL. Under the conditions above, xylanase activity reached 113585.78（U/mL) when FJAT-14260 was fermented for 32h，which was 4.62 folds of that befor optimization. Key words:Bacillus sp；xylanase；fermentation conditions 木聚糖是多聚五碳糖其主链是由 β-1, 4糖苷键将多个D-木糖残基连接起来的[1]，是主要存在于植物细胞壁中的半纤维素成分，在自然界中的含量丰富来源广泛，除纤维素以外，是自然界中最主要的可再生资源多糖[]。木聚糖广泛的存在与植物体内，然而在动物的消化道内基本不能被消化吸收，这样会让一些营养物质无法被吸收利用，很强的抗营养性就表现出来，因此限制了很多含有木聚糖植物饲料在饲料工业和畜牧业生产中的应用[]。 木聚糖酶是一类重要的木聚糖甘键水解酶，也是一种诱导酶，主要包括1,4-β-木聚糖内切酶、1,4-β-D木糖