摘要
目的采用人工神经网络(artificial neural network,ANN)优化一种可获得高生物量及菌体活力大肠埃希菌(E.coli)的摇瓶培养基。方法以葡萄糖(glucose,Glu)、酵母浸出物(yeast extract,YE)、酵母蛋白胨(yeast peptone,YP)、大豆蛋白胨(soy peptone,SP)和酵母基础氮源(yeast nitrogen base,YNB)占比为混料分量,菌体悬浮液A_(600)(A1)、培养物湿菌重(G,g/L)和菌体活力指标值(A2,A_(460))为响应值,采用混料设计试验筛选对响应值具有显著影响的混料分量。以混料设计试验结果为训练和验证数据样本构建ANN模型,输入变量为混料分量且约束同混料分量上下限,输出变量为混料设计响应值。通过所构建ANN获得的优化后培养基配方和参考值,应用蒙特卡洛模拟进一步调整培养基配方,并获得E.coli摇瓶培养基配方,再对培养基配方进行10次验证试验。结果E.coli摇瓶培养基配方:Glu 26 g/L、SP26 g/L、YNB 13 g/L,培养基总浓度为65 g/L。验证结果为:培养可获得A1≥14的批次几率为60%,G≥77 g/L的批次几率为50%,A2≥11的批次几率为40%。培养结果数据均值与参考值均等价。结论本研究优化获得的E.coli摇瓶培养基可培养获得高生物量及菌体活力的E.coli。
Objective To optimize a shake flask culture medium for Escherichia coli(E.coli)with high biomass and viability using artificial neural networks(ANN).Methods Using the proportion of glucose(Glu),yeast extract(YE),yeast peptone(YP),soy peptone(SP)and yeast nitrogen base(YNB)as the mixture component,and the A_(600)(A1)value of cell suspension,wet bacterial weight(G,g/L)of culture and cell viability(A2,A_(460))as the response values,the mixture design was used to screen the mixture components that had a significant effect on the response value.The ANN model was constructed with the test results of mixture design as training and verification data samples.The input variables were mixture components and restricted the upper and lower limits of the mixture components,and the output variables were mixture design response values.The optimized medium formula and reference values were obtained by the constructed ANN.The medium formula was further adjusted by Monte Carlo simulation to obtain the shake flask medium formula of E.coli,which was then verified for 10 times.Results The shake flask culture medium of E.coli was composed of Glu 26 g/L,SP 26 g/L,YNB13 g/L with the total concentration of 65 g/L.The verification results showed that the probability of A1≥14 was 60%,the probability of G≥77 g/L was 50% and the probability of A2≥11 was 40%.The mean values of the incubation result data were equivalent to the reference values.Conclusion The shake flask culture medium of E.coli optimized in this study can obtain E.coli with high biomass and bacterial activity.
作者
徐世友
易中恒
张婷婷
郭济中
张旭东
焦磊
雷清
XU Shiyou;YI Zhongheng;ZHANG Tingting;GUO Jizhong;ZHANG Xudong;JIAO Lei;LEI Qing(Lanzhou Institute of Biological Products Co.,Ltd,Center for Gansu Provincial Vaccine Engineering Research,Lanzhou 730046,Gansu Province,China)
出处
《中国生物制品学杂志》
CAS
CSCD
北大核心
2023年第11期1378-1382,1390,共6页
Chinese Journal of Biologicals
基金
国家重点研发计划(2018YFC1200602)。
关键词
人工神经网络
大肠埃希菌
摇瓶
培养基
Artificial neural network(ANN)
Escherichia coli(E.coli)
Shake flask
Culture medium
作者简介
通信作者:雷清,E-mail:hevlei@sohu.com。
