文献类型：期刊文献

英文题名：Modeling of protein refolding from inclusion bodies

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

机构：[1]Natl Engn Ctr Biochip Shanghai, Shanghai 201203, Peoples R China;[2]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[3]Shandong Econ Univ, Sch Finance, Jinan 250014, Peoples R China;[4]S China Univ Technol, Sch Biosci & Bioengn, Guangzhou 510006, Guangdong, Peoples R China

年份：2009

卷号：41

期号：12

起止页码：1044

外文期刊名：ACTA BIOCHIMICA ET BIOPHYSICA SINICA

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000272463800009)】；

基金：This work was supported by the grants from the National High Technology Research and Development Program of China (No. 2007AA021702), National Basic Research Program of China (No. 2007CB512402), Guangzhou Science and technology Project (No. 2007J1-C0131), Shanghai Rising-star Program (No. 07QB14026), and Shanghai Climbing- Mountain Program (No. 064319039).

语种：英文

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 proteins following dilution refolding 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 properties in the protein refolding. Our results indicated that under the experimental conditions, isoelectric point of proteins might be mostly contributing to the high efficacy 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 (R-2 = 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 refolding developed here could be used to predict the refolding yields of proteins from IBs through a simple method. Our study will be suggestive to optimize the methods for protein refolding from IBs according to their intrinsic properties.