响应面法优化黑曲霉产纤维素酶的发酵条件  

Optimization of Fermentation Conditions for Cellulase Production by Aspergillus niger Using Response Surface Methodology

作　　者：刘松[1] 李祝[1] 周礼红[1] 葛永怡[1] 刘吴娟[1] 张传萍[1] 

机构地区：[1]贵州大学生命科学院,贵州贵阳550025

出　　处：《食品科学》2013年第17期225-229,共5页Food Science

基　　金：毕节循环经济研究院专项[毕循专合字(2010)ZK009号];贵州省农业攻关科技计划项目[黔科合NY字(2011)3053];贵州省科技厅社会发展科技攻关计划项目[黔科合SZ字(2009)3005]

摘　　要：采用单因素试验和响应面法对黑曲霉(Aspergillus niger)xj产纤维素酶的液体发酵条件进行优化并以羧甲基纤维素酶活力作为响应值。首先通过单因素试验初步确定葡萄糖2g/100mL、酵母膏1g/100mL和聚乙二醇体积分数0.1%,初始pH7.0,250mL三角瓶装30mL液体培养基、接种种龄为24h、接种量10%。再利用PlackettBurman(PB)设计筛选出影响羧甲基纤维素酶(CMCase)活力的显著因素:接种量和碳源。通过最陡爬坡试验逼近最大酶活力区域。最后用中心复合设计及响应面分析确定产酶的最佳发酵条件为在PDA的基础上,葡萄糖2.7g/100mL、酵母膏1.25g/100mL和聚乙二醇0.1%、种龄30h、装液量37.5mL(250mL三角瓶)、接种量8%(3.3mL)、初始pH7.0、28℃、150r/min培养96h。经过优化,CMCase活力最高为(63.5±1.7)U/mL,比未优化的条件(在PDA的基础上,种龄24h、装液量30mL(250mL三角瓶)、接种量10%(3mL)、葡萄糖2g/100mL、初始pH7.0、28℃、150r/min培养96h)得到的酶活力最高值(49.0±1.7)U/mL提高了29.59%。Response surface methodology was used to optimize the fermentation conditions for cellulase production by Aspergillus niger to obtain the maximum carboxymethyl cellulase （CMCase） activity. By one-factor-at-a-time designs, we found the optimum fermentation conditions to be inoculum age of 24 h and incoculum amount of 10%, and 30 mL of culture medium composed of 2 g/100 mL glucose, 1 g/100 mL yeast extract and 0.1% PEG at initial pH 7.0 filled in a 250-mL triangular flask. Inoculum amount and glucose concentration were identified by Plackett-Burman （PB） design as main factors that influence CMCase activity, and their optimal regions were approached by the steepest ascent path. Finally, using central composite design and response surface analysis, the optimal fermentation medium was determined as 37.5 mL of a modified PDA medium consisting of 2.7 g/100 mL glucose, 1.25 g/100 mL yeast extract and 0.1 g/100 mL PEG in a 250-mL triangular flask, and the optimal fermentation conditions were 8% （3.3 mL）, 7.0, 28 ℃, 150 r/min and 96 h for inoculums size, initial pH, temperature, shaking speed and time, respectively. The maximum CMCase activity under the optimized conditions was （63.5 ± 1.7） U/mL, indicating a 29.59% increase compared to （49.0 ± 1.7） U/mL obtained under the conditions of 30 mL of PDA with 2% glucose at initial pH of 7.0 in a 250-mL triangular flask, 10% （3 mL） inoculum size with an age of 24 h, and culture at 28 ℃ for 96 h with a shaking speed of 150 r/min.

关 键 词：纤维素酶 液体发酵 黑曲霉 优化 响应面法 

分 类 号：S476.1[农业科学—农业昆虫与害虫防治]