李春梅‘何江波2陈惠2‘王红宁3韩学易2吴琦2
1四川农业人学农学院，成都610064;2四川农业人学生命科学与理学院，雅安625014;3四川人学生命科学学院动物疾病防控生物工程研究
中心，成都610064
*通讯作者，chen62hui@yahoo.com.cn
摘要为了加快纤维素酶土业化应用的步伐，解决纤维素酶发酵土业产量低、生产成本高等问题，本研究
采用正交设计法对高产中性纤维素酶的巨大芽胞杆菌姆acallus megateraum)基因土程菌的培养基和发酵条件
进行优化。结果表明，最佳发酵培养基成分为:麦鼓2.5%、玉米浆1.5%,磷酸二氢钾0.75%、硫酸镁0.04%,氯
化钠0.25% >最佳发酵条件为:温度37 0C , pH 7.0,罐压0.030.05 Mpa，转速600 r/min，装液量3L，接种量
10%，培养4h后以0.25 g/L/h的流速流加木糖lOh，发酵16h后，以恒溶氧方式流加补料8h，共补加料320
mL，发酵过程控制溶氧浓度>_ 30%。该土程菌在此最佳发酵条件下培养，纤维素酶活力可达3846.48 U/mL}
是优化前摇瓶发酵的4.3倍。本研究可为土业化生产纤维素酶提供依据。
关键词纤维素酶，巨大芽胞杆菌，培养基，发酵条件，正交设计
Optimization of Fermentation Conditions for Producing Neutral Cellulase
from a High-yield Bacillus megaterium Genetic Engineering Bacteria
Li Chunmei' He Jiangbo' Chen Hui'-} Wang Hongning3 Han Xueyi'- Wu Qi'-
1 College of Agronomy, Sichuan Agricultural University, Chengdu 610064, China; 2 College of Life and Physical Sciences, Sichuan Agricultural
University, Ya'an 625014, China; 3 Bioengineering Research Center for Animal Disease Prevention and Control, College of Life Sciences, Sichuan
University, Chengdu 610064, China
*Corresponding author, chen62hui@yahoo.com.cn
DOI: 10.3969/j.issn.1674-7968.2011.03.023
Abstract To speed up the pace of industrial application of cellulose, and solve the low yield, high production
cost issues in fermentation Indus抑，orthogonal design methodology was applied to optimize the fermentation of
Bacallus megazeraum genetic engineering bacteria for neutral cellulase. Results indicated that optimal medium
compositions were 2.5% wheat bran, 1.5% corn thick liquid, 0.75% KH}P04, 0.04% MgS04, and 0.25% NaCl; The
optimal fermentation conditions were fermentation temperature of 370C，pH 7.0, tank pressure 0.030.05 Mpa,
rotation speed of shaker 600 r/min, 3 L of packing volume, 10% inoculation amount, fed 10 h xylose场0.25 g/L/h
flow rate after 4 h cultured, constant dissolved oxygen fed-batch 8 h after 16 h of fermentation, dissolved oxygen>_
30% during the fermentation process. The activity of neutral cellulase obtained at this optimal fermentation
condition was 3 846.48 U/mL, which was 4.3 times of the shaking flask culture. This research can provide the
basis for the cellulase manufacture.
Keywords Cellulase, Bacallus ma}-teraum, Medium, Fermentation conditions, Orthogonal design
