粗糙脉孢菌(<em>Neurospora crassa</em>)纤维素酶液体发酵培养基的优化

中国生物工程杂志

2012, Vol. 32

Issue (03): 76-82

技术与方法

粗糙脉孢菌(Neurospora crassa)纤维素酶液体发酵培养基的优化

顾芮萌^1,2, 李勇昊^2,3, 田朝光²

1. 天津科技大学食品工程与生物技术学院天津 300457;
2. 中国科学院天津工业生物技术研究所天津 300308;
3. 吉林大学生物与农业工程学院长春 130022

The Medium Optimization of Cellulases Fermentation of Neurospora crassa by Response Surface Methodology

GU Rui-meng^1,2, LI Yong-hao^2,3, TIAN Chao-guang²

1. College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China;
2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China;
3. Bioengineering and Agricultural Engineering College, Jilin University, Changchun 130022, China

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摘要：

粗糙脉孢菌是天然纤维素降解真菌,具有产纤维素酶能力,国内外对其纤维素降解机理和发酵产酶有一定的研究,但对其产酶的条件优化研究得不多,其产酶潜力需要进一步挖掘。以粗糙脉孢菌基因组测序菌株FGSC 2489为对象,采用响应面分析法对Neurospora crassa摇瓶发酵产纤维素酶进行培养基优化。采用Plackett-Burman(PB)实验设计考察发酵培养基中关键参数对产酶条件的影响,进而采用最陡爬坡实验逼近最大响应区域,并结合中心组合实验(central composite design, CCD)和响应面分析法对两个显著因素进行分析。PB实验结果显示:Peptone、Yeast extract对产纤维素酶有显著影响。通过响应面分析得到一元二次方程,对方程求解得到Peptone 7.27g/L、Yeast extract 5.51g/L。采用该优化培养基,最大纤维素酶活可达1.27FPU/ml, 较优化前提高了2.03倍;CMC酶活 14.15IU/ml,比优化前提高1.88倍;木聚糖酶活24.13IU/ml,比优化前提高1.86倍;葡萄糖苷酶酶活1.22IU/ml 比优化前提高2.08倍。

关键词： 粗糙脉孢菌; 响应面; 纤维素酶

Abstract:

Neurospora crassa is the native degrader of cellulose, it has capability to produce the mainly components of cellulases, such as CBHs, endoglucanses and β-glucosidases. It has some studies focus on the mechanism or cellulases productivity in Neurospora crassa, but the cellulases fermentation by this fungus is far from optimized. Here, The Response Surface Methodology(RSM) to optimize the medium for cellulased production in FGSC.2489. Combined the Plackett-Burman and Central Composite Design design, the peptone, yeast extract have significant effects on enzyme activity were found. The optimum amount of the Peptone is 7.27g/L and Yeast extract is 5.51g/L in medium. Based on the optimized medium, the cellulases production have significantly improved near 2 folds, from 0.64FPU to 1.27 FPU.

Key words: Neurospora crassa Response surface methodology Cellulases

收稿日期: 2011-11-03 出版日期: 2012-03-25

ZTFLH:

Q815

基金资助:

国家"973"计划资助项目(2011707403)

通讯作者: 田朝光,tian_cg@tib.cas.cn E-mail: tian_cg@tib.cas.cn

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引用本文:

顾芮萌, 李勇昊, 田朝光. 粗糙脉孢菌(Neurospora crassa)纤维素酶液体发酵培养基的优化[J]. 中国生物工程杂志, 2012, 32(03): 76-82.

GU Rui-meng, LI Yong-hao, TIAN Chao-guang. The Medium Optimization of Cellulases Fermentation of Neurospora crassa by Response Surface Methodology. China Biotechnology, 2012, 32(03): 76-82.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I03/76

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