摘要
目的:筛选合成培养基中影响重组毕赤酵母高效表达木聚糖酶的关键成分。方法:首先通过单因素实验和t检验筛选重组毕赤酵母表达木聚糖酶的最佳初始诱导培养基,其次利用Plackett-Burman设计及逐步回归建立培养基成分和响应值间的多元线性回归模型,并筛选出关键培养基成分,最后利用相关系数分析,对非关键成分含量的选取做出合理的取舍。结果:筛选出甘油磷酸钠、硫酸钙、PTM1和硫酸铵是影响重组酵母表达木聚糖酶的关键培养基成分,且当培养基组合为甘油磷酸钠20 g/L,(NH4)2 SO42.0 g/L,CaSO4·2H2O 2.0 g/L,K2 SO420.0 g/L,MgSO4·7H2O 6.0 g/L,PTM18.0 mL/L,甲醇10.0 mL/L,吐温802.0 g/L时,木聚糖酶的酶活力和比酶活分别达到2298.4 U/mL和9926.3 U/mg,分别是优化前的3.055倍和3.889倍。结论:Plackett-Burman设计不仅可以显著提升培养基优化目标,还能筛选出关键培养基成分,从而为进一步优化奠定了基础。
Objectives:The aim of this study was to screen the key components in synthetic medium that affect the efficient expression of xylanase in recombinant Pichia pastoris.Methods:Firstly,the best original defined medium was selected by employing single factor method and t-test.Secondly,the key components were screened via establishing the multivariate linear regression models between medium components and responses with Plackett-Burman design and stepwise regression.Finally,the concentration of non-critical medium components were determined with correlation coefficient analysis.Results:Glycerophosphate,calcium sulfate,PTM1 and ammonium sulphate were screened as key components.Furthermore,it was found that the enzyme activity and specific enzyme activity of xylanase reached maximum values of 2298.4 U/mL and 9926.3 U/mg in Plackett-Burman design,respectively(i.e.when glycerophosphate 20 g/L,(NH4)2 SO42.0g/L,CaSO4·2H2O 2.0 g/L,K2 SO420.0 g/L,MgSO4·7H2O 6.0 g/L,PTM18.0 mL/L,Methanol 10.0 mL/L,Tween 802.0 g/L),which were 3.055 and 3.889 times as many as before optimization,respectively.Conclusion:Plackett-Burman design not only significantly enhance the optimization objectives,but also screen the key components,and thus lay the foundation for further optimization.
作者
朱新术
赵文慧
张晓蕾
林波
刘超
张苗
陈菲
ZHU Xin-shu;ZHAO Wen-hui;ZHANG Xiao-lei;LIN Bo;LIU Chao;ZHANG Miao;CHEN Fei(College of Medical Technology,Jiangsu College of Nursing,Huai’an 223005,China;Department of Basic Medicine,Jiangsu College of nursing,Huai’an 223005,China)
出处
《食品工业科技》
CAS
北大核心
2020年第7期109-114,共6页
Science and Technology of Food Industry
基金
江苏护理职业学院高层次人才科研启动基金(SHKYQD2018003)
淮安市科技局自然科学研究计划(HAB201844)。
作者简介
朱新术(1980-),男,博士,副教授,研究方向:微生物发酵工艺优化与代谢组学,E-mail:xinshu0518@163.com;通讯作者:赵文慧(1980-),女,博士,讲师,研究方向:神经损伤与保护,E-mail:wenhui8088@gmail.com。
