片烟黑曲霉菌株产纤维素酶的条件优化及酶的分离纯化

›› 2012, Vol. 24 ›› Issue (6): 0-1128.

• 论文 •

片烟黑曲霉菌株产纤维素酶的条件优化及酶的分离纯化

范坚强1，周彬2，宋纪真3，王金华2，*，陈义强1，王永泽2

1福建中烟工业有限责任公司技术中心，福建厦门 361022；2湖北工业大学生物工程学院，湖北武汉 430068； 3郑州烟草研究院，河南郑州 450001

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2012-11-25 发布日期:2012-11-25

Optimization of cellulase production by Aspergillus niger separated from tobacco and its separation and purification

FAN Jianqiang;ZHOU Bin;SONG Jizhen;WANG Jinhua;*;CHEN Yiqiang;WANG Yongze

1 Technology Center， Fujian Branch of China Tobacco Industry Co， Ltd， Xiamen 361022， China； 2 School of Biotechnology， Hubei University of Technology， Wuhan 430068， China；3 Zhengzhou Tobacco Research Institute of CNTC， Zhengzhou 450001， China

Received:1900-01-01 Revised:1900-01-01 Online:2012-11-25 Published:2012-11-25

摘要/Abstract

摘要： 为降低烟叶燃吸的刺激味、改善口感、增加香气、提高烟叶品质，从烟叶表面筛选到一株产纤维素酶活性较高的黑曲霉，应用PlackettBurman试验确定4种显著因子，RSM法对4种显著因子进行了优化和分析，并对酶蛋白的分离纯化作了初步研究。结果显示：发酵产酶的最适碳源为玉米芯，氮源为NH4NO3，当玉米芯用量为13.4%，NH4NO3为0.4%，温度为30.4℃，pH为5.6时产酶比活力最高，可达40.02 U·mL-1。纤维素酶酶蛋白的硫酸铵盐析最佳饱和度为60%，经Sephadex G100凝胶柱层析纯化后的比活力提高了3.5倍，回收率为32.6%，达到较高水平。

关键词: 黑曲霉, 纤维素酶, 响应曲面法, 凝胶电泳

Abstract: In order to improve tobacco taste， decrease the smoking pungent taste， increase the fragrance and improve the smoking quality， a mold strain Aspergillus niger possessing relatively higher cellulase enzymatic activity was isolated from tobacco leaves The PlackettBurman design was used to choose the most important factors， then， these factors were optimized by RSM method， and the purification of enzyme was investigated The results indicated that the optimum carbon source and nitrogen source were 1.4% corn cob and 0.4% NH4NO3， the optimum original pH was 5.6 and the optimum cultural temperature was 30.4℃ Under these conditions， the specific enzyme activity was 40.02 U·mL-1 The crude cellulase was isolated and purified by means of ammonium sulfate precipitation， followed by chromatography on Sephadex G100 The specific activity of the cellulase increased to 3.5 fold of the source enzyme with an activity recovery of 32.6%.

Key words: Aspergillus niger, cellulase, response surface methodology, electrophoresis

范坚强;周彬;宋纪真;王金华;*;陈义强;王永泽. 片烟黑曲霉菌株产纤维素酶的条件优化及酶的分离纯化[J]. , 2012, 24(6): 0-1128.

FAN Jianqiang;ZHOU Bin;SONG Jizhen;WANG Jinhua;*;CHEN Yiqiang;WANG Yongze. Optimization of cellulase production by Aspergillus niger separated from tobacco and its separation and purification[J]. , 2012, 24(6): 0-1128.

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片烟黑曲霉菌株产纤维素酶的条件优化及酶的分离纯化

Optimization of cellulase production by Aspergillus niger separated from tobacco and its separation and purification

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