Optimization of fermentation condition of Pichia pastoris for xylanase production by response analysis
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摘要: 通过单因素实验研究了诱导温度、种龄、p H、甲醇浓度以及诱导时间对毕赤酵母工程菌产木聚糖酶的影响。在此基础上进行响应面优化设计实验,并根据结果拟合小二乘二次项回归方程,探讨了各因素对木聚糖酶比酶活力的影响。确定了最优的发酵培养条件:种龄为31 h,诱导时间104 h,甲醇诱导浓度1%,发酵起始p H4.0,诱导温度为30℃,在此条件下对实验结果进行验证,得到木聚糖酶比酶活力为43526.3 U/mg。Abstract: The influence of the temperature,inoculum time,p H,methanol concentration and induction time on the xylanase production of pichia recombinant bacteria were studied. The response surface optimazation was designed on the basis of the single factor experiment,and the small squares quadratic regression equation was fitted out according to the results to discuss the influence of the various factors on the xylanase enzyme activity. The optimum fermentation culture conditions were as follows:inoculum age 31 h,induction time 104 h,methanol concentration induced by 1%,fermentation starting p H4.0,induction temperature 30 ℃. On this condition,the result of the experiment for xylanase enzyme activity was 43526.3 U/mg.
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Keywords:
- Pichia pastoris /
- xylanase /
- fermentation conditions /
- response surface
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[1] 吴萍,李正鹏,何庆元,等.金针菇固态发酵产木聚糖酶的纯化及其性质研究[J].药用生物技术,2012,19(1):31-34. [2] 符丹丹,李爱江,谢慧,等.宇佐美曲霉木聚糖酶的纯化及其性质[J].食品科学,2006,27(2):116-120. [3] Coughlan M P,Hazlewood G P.β-1,4-D-xylan-degrading enzyme system:biochemistry,molecularbiology a nd applications[J].Biotechnol Apply Bioche,1993,17:259.
[4] 孙振涛,赵祥颖,刘建军,等.微生物木聚糖酶及其应用[J].生物技术,2007,17(2):93-97. [5] 刘波,邬应龙,张霞,等.红曲霉固态发酵产木聚糖酶培养基的响应面优化[J].食品工业技,2014,35(1):254-258. [6] Bajaj B K,Manhas K.Production and characterization of xylanase from Bacillus licheniformis P11(C)with potential for fruit juice and bakery industry[J].Biocatalysis and Agricultural Biotechnology,2011,1(4):330-337.
[7] Ko C H,Tsai C H,Tu J,et al.Identification of Paenibacillus sp.2S-6 and application of its xylanase on bio-bleaching[J].International Biodeterioration&Biodegradation,2011,65(2):334-339.
[8] Jiang Z Q,Cong Q Q,Yan Q J,et al.Characterisation of a thermostable xylanase from Chaetomium sp.and its application in Chinese steamed bread[J].Food Chemistry,2010,120(2):457-462.
[9] 曹云鹤,陈小玲,贺平丽,等.硫色曲霉木聚糖酶基因xyn A的克隆、表达及酶学性质分析[J].生物技术通讯,2006,17(6):878-881. [10] Shi H L,Yin X,Wu M C,et al.Cloning and bioinformatics analysis of an endoglucanase gene(Aucel12A)from Aspergillus usamii and its functional expression in Pichia pastoris[J].J Ind Microbiol Biotechnol,2012,39:347-357.
[11] 周薇薇,尹亚辉,赵长新.响应面法优化黑曲霉固态发酵产木聚糖酶工艺[J].大连工业大学学报,2013,32(3):176-179. [12] 汤文晶,谢志皓,张庆庆,等.匍枝根霉液态发酵产木聚糖酶培养基优化研究[J].安徽工程大学学报,2014,29(1):1-5. [13] Zhang H L,Yao B,Wang Y R.Expression of xylanase gene xyn A from Stretomyces olivaceoviridis A1 in Escherichia coli and Pichia pastoris[J].China Biotechnology,2003,19(1):76-80.
[14] Yao D,Wang Y,Yang J,et al.Optimization of extraction process for xylan from corncob by response surface methodology[J].Food Science,2011,32(8):111-115.
[15] Bailey M J,Biely P,Poutanen K.Interlaboratory testing of methods for assay of xylanase activity[J].Journal of Biotechnology,1992,23:257-270.
[16] 董悦安.温度变化对地下水中微生物影响的研究[J].勘察科学技术,2008(2):15-18. [17] 王涛,万红贵,蔡恒,等.毕赤酵母发酵产木聚糖酶条件的研究[J].中国酿造,2009,20(8):86-89. [18] 周晨妍.宇佐美曲霉木聚糖酶基因的克隆、表达及定向诱变研究[D].无锡:江南大学,2008.
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