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[1]夏玉林 冯 娟 李 荷.响应面法优化漆酶基因lac1338表达漆酶的发酵条件[J].应用与环境生物学报,2016,22(02):219-223.[doi:10.3724/SP.J.1145.2015.06023]
 XIA Yulin,FENG Juan & LI He**.Optimizing fermentation conditions for the expression of laccase gene lac1338by response surface methodology*[J].Chinese Journal of Applied & Environmental Biology,2016,22(02):219-223.[doi:10.3724/SP.J.1145.2015.06023]
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响应面法优化漆酶基因lac1338表达漆酶的发酵条件()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
22卷
期数:
2016年02期
页码:
219-223
栏目:
研究论文
出版日期:
2016-04-25

文章信息/Info

Title:
Optimizing fermentation conditions for the expression of laccase gene lac1338by response surface methodology*
作者:
夏玉林 冯 娟 李 荷
广东药科大学基础学院生物化学与分子生物学系 广州 510006
Author(s):
XIA Yulin FENG Juan & LI He**
Department of Biochemistry &Molecular Biology, Guangdong Pharmaceutical University, Guangzhou 510006, China
关键词:
漆酶Plackett-Burman响应面发酵条件优化回归方程
Keywords:
laccase Plackett-Burman response surface methodology optimization of fermentation condition regressionequation
分类号:
TQ925
DOI:
10.3724/SP.J.1145.2015.06023
摘要:
为提高Escherichia coli BL21(DE3)中漆酶基因lac1338的表达水平,采用响应面法对其发酵条件进行优化. 首先用Plackett-Burman法筛选出3个影响较大的重要因素,分别为温度(Q1)、诱导时间(Q3)以及诱导前菌体浓度OD600 nm(Q4). 继而结合响应面分析,建立以漆酶酶活为响应值的二次回归方程模型,即Y = 75.33 + 3.30A + 9.35B + 5.35C -1.69AB + 0.80AC + 11.50BC - 25.97A2 + 1.83B2 - 4.66C2,从中获得最优的发酵条件:诱导温度为31 ℃,诱导时间为20 h,诱导前的菌体浓度OD600 nm值为1.8. 采用该发酵条件,供试菌株的漆酶比酶活达到22.8 U/mg,较优化前漆酶的表达量(10.7 U/mg)提高了2.13倍,其试验值与预测值基本相符. 说明预测模型可靠性高,可应用于漆酶发酵条件的优化.(图4 表5 参25)
Abstract:
In order to enhance the production of laccase in Escherichia coli BL21 (DE3), response surface methodology wasapplied to optimize the fermentation conditions. Plackett-Burman design was used to investigate the main factors affectinglaccase yield of E. coli BL21 (DE3). The results showed that three factors played important roles in the fermentation conditions:the culture temperature (Q1), culture time (Q3) and the initial bacteria density of OD600 nm (Q4). The second-order regressionequation model for the yield of laccase was established by response surface analysis as Y=75.33 + 3.30A + 9.35B + 5.35C- 1.69AB + 0.80AC + 11.50BC - 25.97A2 + 1.83B2 - 4.66C2. Based on this model, the optimal fermentation conditions wereobtained as 20 h of culture time, 31 °C of culture temperature, and OD600 nm 1.8 of initial bacteria density. The ferment conditionoptimization increased the laccase production by nearly 2.13 times to 22.8 U/mg, with the experimental values in agreementwith the predicted ones. These results suggested that the prediction model is reliable and suitable for the optimization of laccasefermentation conditions.

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更新日期/Last Update: 2016-04-25