文献类型：期刊文献

中文题名：Modeling of protein refolding from inclusion bodies

英文题名：Modeling of protein refolding from inclusion bodies

作者：Ting Zhang[1];Xiaojing Xu[2];Liang Shen[3];Yanye Feng[1];Zhong Yang[1];Yaling Shen[1];Jufang Wang[4];Weirong Jin[2];Xiaoning Wang[1,4]

机构：[1]The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China;[2]National Engineering Center for Biochip at Shanghai, Shanghai 201203, China;[3]School of Finance, Shandong Economic University, Jinan 250014, China;[4]School of Biosciences and Bioengineering, South China University of Technology, Guangzhou 510006, China

年份：2009

卷号：41

期号：12

起止页码：1044

中文期刊名：Acta Biochimica et Biophysica Sinica

外文期刊名：生物化学与生物物理学报（英文版）

收录：CSTPCD;;Scopus;CSCD:【CSCD2011_2012】；PubMed;

语种：英文

中文关键词：Escherichia coli; inclusion body;statistical modeling; protein refolding

外文关键词：Escherichia coli; inclusion body;statistical modeling; protein refolding

摘要：Overexpression of foreign proteins in Escherichia coli often leads to the formation of inclusion bodies （IBs）, which becomes the major bottleneck in the preparation of recombinant proteins and their applications. In the present study, 36 proteins from IBs were refolded using a simple refolding method. Refolding yields of these proteins were defined as the percentage of soluble pro- teins following dilution refoiding in the amount of denatured proteins in the samples before diluting into refolding buffer. Furthermore, a mathematical model was deduced to evaluate the role of biochemical proper- ties in the protein refolding. Our results indicated that under the experimental conditions, isoelectric point of proteins might be mostly contributing to the high effi- cacy of protein refolding since the increment of one unit resulted in a decrease of 14.83% in the refolding yield. Other important mediators were components of protein secondary structure and the molecular weight （R2= 0.98, P = 0.000, F-test）. Six proteins with low efficiency in the protein refolding possessed relatively low isoelectric points. Furthermore, refolding yields of six additional proteins from IBs were predicted and further validated by refolding the proteins under the same conditions. Therefore, the model of protein refold- ing developed here could be used to predict the refold- ing yields of proteins from IBs through a simple method. Our study will be suggestive to optimize the methods for protein refoiding from IBs according to their intrinsic properties.
Overexpression of foreign proteins in Escherichia coli often leads to the formation of inclusion bodies （IBs）, which becomes the major bottleneck in the preparation of recombinant proteins and their applications. In the present study, 36 proteins from IBs were refolded using a simple refolding method. Refolding yields of these proteins were defined as the percentage of soluble pro- teins following dilution refoiding in the amount of denatured proteins in the samples before diluting into refolding buffer. Furthermore, a mathematical model was deduced to evaluate the role of biochemical proper- ties in the protein refolding. Our results indicated that under the experimental conditions, isoelectric point of proteins might be mostly contributing to the high effi- cacy of protein refolding since the increment of one unit resulted in a decrease of 14.83% in the refolding yield. Other important mediators were components of protein secondary structure and the molecular weight （R2= 0.98, P = 0.000, F-test）. Six proteins with low efficiency in the protein refolding possessed relatively low isoelectric points. Furthermore, refolding yields of six additional proteins from IBs were predicted and further validated by refolding the proteins under the same conditions. Therefore, the model of protein refold- ing developed here could be used to predict the refold- ing yields of proteins from IBs through a simple method. Our study will be suggestive to optimize the methods for protein refoiding from IBs according to their intrinsic properties.