文献类型：期刊文献

英文题名：Efficient synthesis of amino acid polymers for protein stabilization

作者：Li, Bing[1,2,3];Wu, Yueming[1,2,3];Zhang, Wenjing[1,2,3];Zhang, Si[1,2,3];Shao, Ning[1,2,3];Zhang, Weiwei[1,2,3];Zhang, Lixin[1];Fei, Jian[4];Dai, Yidong[5];Liu, Runhui[1,2,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Key Lab Ultrafine Mat, Minist Educ, Shanghai 200237, Peoples R China;[3]East China Univ Sci & Technol, Res Ctr Biomed Mat, Minist Educ, Shanghai 200237, Peoples R China;[4]Shanghai Jiao Tong Univ, Dept Gen Surg, Sch Med, Ruijin Hosp, Shanghai 200025, Peoples R China;[5]Shanghai Ruijin Rehabil Hosp, Shanghai 200023, Peoples R China

年份：2019

卷号：7

期号：9

起止页码：3675

外文期刊名：BIOMATERIALS SCIENCE

收录：;EI(收录号：20193507378339);WOS:【SCI-EXPANDED(收录号:WOS:000482087100011)】；

基金：This research was supported by the National Natural Science Foundation of China (No. 21774031, 21574038, 21861162010, 31800801), the National Key Research and Development Program of China (2016YFC1100401), the National Natural Science Foundation of China for Innovative Research Groups (No. 51621002), the Natural Science Foundation of Shanghai (18ZR1410300), the "Eastern Scholar Professorship" from Shanghai local government (TP2014034), the national special fund for State Key Laboratory of Bioreactor Engineering, the Fundamental Research Funds for the Central Universities (22221818014), the authors also thank Research Center of Analysis and Test of East China University of Science and Technology for the help on the characterization.

语种：英文

外文关键词：Biosynthesis - High resolution transmission electron microscopy - Ring opening polymerization - Stabilization - Lithium compounds - Proteins - Dynamic light scattering - Phase locked loops

摘要：Proteins are fragile such that even freezing, drying and dehydration may induce their denaturation, aggregation, and activity loss. To protect proteins from these kinds of damage, we prepared two types of amino acid polymers, poly-(l-glutamate)-r-poly-(l-lysine) (PLG-r-PLL) and poly-l-glutamate (PLG), from the efficient ring-opening polymerization of alpha-amino acid N-carboxyanhydride (NCA) using lithium hexamethyldisilazide (LiHMDS) as the initiator. beta-galactosidase (beta-Gal) was used in this study to examine the protein protecting effect of the synthesized amino acid polymers during lyophilization. The results indicate that both PLG-r-PLL and PLG exert significant protection on beta-Gal during lyophilization and improve the activity of the resulting protein from 40%, without using a protecting agent during lyophilization, to 80% of the original protein activity. Nevertheless, PLG generally performs better than PLG-r-PLL independent of the chain length. Our studies also show that PLG and PLG-r-PLL with a high content of PLG subunits display no observable cytotoxicity and hemolytic effect. Furthermore, dynamic light scattering (DLS) and transmission electron microscopy (TEM) characterization indicate that PLG protects beta-Gal upon lyophilization by preventing the aggregation of beta-Gal. Our studies demonstrate that amino acid polymers, such as PLG, can exert potent activity for protein stabilization. The easy operation of LiHMDS-initiated and efficient NCA polymerization implies the great potential of this strategy to prepare amino acid polymers quickly for the screening of protein stabilization and mechanism study.