摘要
目的响应面法优化高压均质破碎重组汉逊酵母细胞的工艺条件。方法以均质压力(A)、均质次数(B)、均质酵母质量分数(C)3个因素为影响因素,酵母细胞破碎率(Y)为响应值,应用Design-Expert软件,根据Box-Behnken中心组合试验设计方案,用响应曲面法建立二阶数学模型,确定最佳破碎工艺参数,基于二次多项式回归模型建立设计空间,并进行规模化工艺验证。结果方差分析结果显示,模型P<0.01,且失拟值>0.05,表明模型与检测结果拟合较好。最终获得的操作空间为:均质压力1125~1200 bar,均质次数3~4次,均质酵母质量分数12%。3批重组汉逊酵母细胞发酵液中酵母细胞破碎率均达到65%以上,该值在响应值设计空间范围内。结论响应面法优化的高压均质破碎重组汉逊酵母细胞的工艺具有较好的稳健性和适应性。
Objective To optimize the technological condition for disruption of recombinant Hansenula polymorpha cells with high pressure homogenization by response surface methodology.Methods Serving the homogeneous pressure(A),homogeneous times(B)and homogeneous yeast mass fraction(C)as independent variables,and the yeast cell fragmentation rate(Y)as response value,Box-Benhnken center-united experiment was used for experimental arrangements,and the homogenization process parameters were optimized by response surface methodology.The design space of homogenization process was established based on the quadratic polynomial regression model,and the large-scale process was verified.Results The results of ANOVA showed that the P value of the model was less than 0.01,while the P value of lack of fit was more than 0.05,which indicated that the model was fitted well with the test results.The recommended operation space of parameters was as follows:homogeneous pressure 1125~1200 bar,homogeneous frequency 3~4 times,homogeneous yeast mass fraction 12%.All the disruption rates of cells in three batches of fermentation liquids of H.polymorpha were more than 65%,which were within the designed space of response value.Conclusion The high homogenization process for disruption of H.polymorpha cells optimized by response surface methodology showed high robustness and flexibility.
作者
张生琰
王玢
邵志伟
陈卓涛
杨林鹏
谢忆
李薇
雷清
杨俊杰
ZHANG Sheng-yan;WANG Bin;SHAO Zhi-wei;CHEN Zhuo-tao;YANG Lin-peng;XIE Yi;LI Wei;LEI Qing;YANG Jun-jie(Pilot Plant Test Lab,Lanzhou Institute of Biological Products Co.,Ltd.,Center for Gansu Provincial Vaccine Engineering Research,Lanzhou730046,Gansu Province,China)
出处
《中国生物制品学杂志》
CAS
CSCD
北大核心
2022年第9期1112-1117,共6页
Chinese Journal of Biologicals
关键词
响应面法
工艺优化
高压均质破碎
重组汉逊酵母细胞
Response surface methodology
Process optimization
High pressure homogenization
Recombinant Hansenula polymorpha cells