范亚苇-南昌大学-响应面法优化粗壮脉纹孢菌CGMCC 3088固态发酵产纤维素酶培养基.pdf

响应面法优化粗壮脉纹孢菌CGMCC 3088 固态发酵产纤维素 酶培养基  张玮 范亚苇 邓泽元 李红艳 李静 （南昌大学食品科学与技术国家重点实验室，江西南昌 330047 ） 摘要：采用Plackett-Burman 设计和响应面法对粗壮脉纹孢菌固态发酵产纤维素酶的培养基组成进行优化。 结果表明：豆皮和酒糟是粗壮脉纹孢菌发酵产纤维素酶的最优碳源。通过 PB 试验筛选出豆皮、酒糟和硫 酸铵的添加量是影响产酶的主要因子，最后通过最陡爬坡试验和中心组合试验优化得出产纤维素酶的最优 培养基组合为：豆皮 64%，酒糟34%，硫酸铵2%，在此培养基下粗壮脉纹孢菌发酵产纤维素酶的酶活为 2.83IU/g ，较优化前的1.17IU/g 提高了141.88%。 关键词：粗壮脉纹孢菌；纤维素酶；Plackett-Burman 设计；响应面；培养基 Optimization of solid state fermentation medium of Neurospora crassa CGMCC 3088 for cellulases by response surface analysis Abstract: Plackett-Burman design and response surface methodology were applied to optimize the components in solid state fermentation medium for cellulases production of Neurospora crassa CGMCC 3088. The result shows that bean hull and distillers ’ grains are the best carbon source on solid state fermentation medium. Plackett-Burman design was applied to determine the specific medium components affecting cellulases activity and found that bean hull ，distillers ’grains and ammonia sulfate were the key factors. Based on these results, steepest ascent experiments and central composite design were used for further optimization. Finally, the optimal medium was: 64% bean hull, 34% distillers ’grains and 2% ammonia sulfate. Under these conditions, the activity of cellulases significantly improved 141.88%, from 1.17IU/g to 2.83IU/g. Key words: Neurospora crassa ; cellulases; Plackett-Burman design; RSM; medium 纤维素酶是一组能降解木质纤维素的复杂酶系，纤维素酶的来源广泛，利用微生物发酵 产纤维素酶是目前纤维素酶生产的主流。 本实验使用的粗壮脉纹孢菌，是由实验室自行筛选并诱变育种的一株高产纤维素酶的菌 [1] 株 。它不仅具有健全的纤维素酶系，而且在高纤维含量的培养基上生长旺盛。由于纤维素 [2] 酶的调节因 ace2 只受纤维素的影响 ，所以不同纤维原料的纤维组成和结构不同诱导生产 [3-5] 纤维素酶的活性差异较大 。本实验选取豆皮、豆渣、米糠、啤酒糟、酒糟5 种不同农副 产品为纤维底物，对比不同纤维对真菌发酵产纤维素酶的影响，寻找最适