第27卷第1期高校化学工程学报No.1 V ol.27 2013 年2 月 Journal of Chemical Engineering of Chinese Universities Feb. 2013文章编号：1003-9015(2013)01-0113-06
产纤维素酶增效蛋白菌株的筛选及培养条件优化
覃益民1,2, 陶恒1, 刘幽燕1,2, 李青云1,2
(1. 广西大学化学化工学院, 广西南宁530004; 2. 广西生物炼制重点实验室, 广西南宁530003)
摘要：为了寻找高效的产纤维素酶增效蛋白菌株，建立一种筛选方法从土壤中筛选得到一株产纤维素酶增效蛋白的
细菌POEP1，其所产增效蛋白对纤维素酶(来源于黑曲霉Aspergillus niger)的滤纸酶活有明显的增效作用，通过对其进行
16SrDNA分类鉴定，确定该菌株为栖稻假单胞菌(Pseudomonas oryzihabitans)。为进一步提高POEP1产纤维素酶增效蛋
白的水平，对其培养条件进行优化。采用单因子试验筛选出最佳碳源为麦麸，最佳氮源为黄豆粉。再通过Plackett-Burman
设计对与发酵相关的11个因子进行试验，筛选出麦麸、黄豆粉和KH2PO4为3个主要显著因子；由Box-Behnken 实验
结合响应面分析确定主要因子的最优水平为：麦麸质量浓度32.3 g?L?1，黄豆粉质量浓度24.7 g?L?1，KH2PO4质量浓
度 4.36 g?L?1。优化后纤维素酶增效蛋白的产量提高1.7倍，增效活力由15.79 U?mL?1增至43.31 U?mL?1。
关键词：纤维素酶增效蛋白；Plackett-Burman 设计；响应曲面；发酵优化
中图分类号：TQ925.9；Q556.9 文献标识码：A DOI：10.3969/j.issn.1003-9015.2013.01.019 Screening of the Cellulase-Enhancing-Protein Producing Strain and
Optimization of its Culture Conditions
QIN Yi-min1,2, TAO Heng1, LIU You-yan1,2 , LI Qing-yun1,2
(1. School of Chemistry and Chemical Engineering , Guangxi University, Nanning 530004, China;
2. Guangxi key laboratory of biorefinery, Nanning 530003, China)
Abstract: A self-designed screening model was built for the screening of the high-efficient cellulase- enhancing-protein producing strains. A strain (named POEP1), having the ability of producing cellulase-activity-enhancing-protein, was isolated from the soil, and then according to the 16S rDNA gene sequencing analysis, the strain was identified to be Pseudomonas oryzihabitans. Some fermentation conditions were optimized to increase the protein productivity. The most suitable carbon and nitrogen sources were determined to be wheat bran and soybeans meal in the single-factor experiments, respectively. Three key factors such as mass concentrations of wheat bran, soybeans meal and KH2PO4 were selected from eleven factors affecting the protein productivity by Plaekett-Burman design and were further optimized by Box-Behnken design and response surface methodology. The optimal concentrations of these three important factors were obtained as follows 32.3 g?L?1 of wheat bran, 24.7 g?L?1 of soybeans meal and 4.36 g?L?1of KH2PO4. Under the optimal conditions，the cellulase-enhancing activity of 43.31 U?mL?1 was obtained, which is 1.7 times higher than that before optimization (15.79 U?mL?1) .
Key words: cellulase-enhancing-protein; Plackett-Burman design; response surface methodology;
fermentation optimization
1引言
纤维素是最丰富的天然可再生生物资源，利用纤维素酶水解纤维素生产燃料乙醇越来越引起人们的关注[1~3]。然而纤维素酶的水解效率低是制约纤维素生产燃料乙醇的瓶颈。
收稿日期：2011-11-22；修订日期：2012-04-17。
基金项目：国家自然科学基金(21066001)；广西科学基金资助项目(桂科自0991031)。
作者简介：覃益民(1962-)，男，广西贵港人，广西大学副教授，博士。通讯联系人：刘幽燕，E-mail：liuyouyangx@https://www.doczj.com/doc/f08464546.html,