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作 者:李冰[1] 丛自豪 肖希勉 武月铭 刘润辉[1] LI Bing;CONG Zihao;XIAO Ximian;WU Yueming;LIU Runhui(State Key Laboratory of Bioreactor Engineering,Key Laboratory for Ultrafine Materials of Ministry of Education,Research Center for Biomedical Materials of Ministry of Education,School of Materials Science and Engineering,East China University of Science and Technology,Shanghai 200237,China)
机构地区:[1]华东理工大学生物反应器工程国家重点实验室,超细材料制备与应用教育部重点实验室,教育部医用生物材料工程研究中心,材料科学与工程学院,上海200237
出 处:《华东理工大学学报(自然科学版)》2020年第4期526-532,共7页Journal of East China University of Science and Technology
基 金:国家自然科学基金(21774031,21574038)。
摘 要:为提高蛋白在冻干过程中的稳定性,采用双(三甲基硅基)氨基锂(LiHMDS)引发的超快速氨基酸环内酸酐(NCA)开环聚合方法,制备了聚L-赖氨酸(PLL)均聚物和聚L-谷氨酸(PLG)均聚物。以β-半乳糖苷酶(β-Gal)为模型蛋白检测氨基酸均聚物在冻干过程中对蛋白的保护效果,结果表明两种氨基酸均聚物的共混物对蛋白有保护效果,冻干后未添加氨基酸共混物保护剂的蛋白活性仅剩38%,添加氨基酸共混物保护剂的蛋白活性明显提升到77%,表明氨基酸共聚物能够在冻干过程中保护蛋白,显著提高蛋白稳定性。本文方法操作简单,可快速合成氨基酸聚合物并高效筛选蛋白稳定剂。Proteins are widely used as reagents in laboratories and as therapeutics for a variety of diseases.However,the native proteins are prone to aggregation and can be denatured under extreme conditions such as freezing,drying and dehydration,leading to a reduced activity,and thus an increased dosage.To achieve protein protection under extreme conditions,we prepared two types of amino acid polymers,poly-(L-lysine)(PLL)and poly-L-glutamate(PLG)from the rapid ring-opening polymerization(ROP)ofα-amino acid N-carboxyanhydride(NCA)using lithium hexamethyldisilazide(LiHMDS)as the initiator.β-galactosidase(β-Gal)was used as a model protein to examine the protein protective effect of the synthesized amino acid polymers during lyophilization.The results showed that the presence of PLL and PLG blending had significant protection effect on β-Gal activity during lyophilization.The enzyrnatic activity of the lyophilized protein was improved from 38%(without protecting agent)to 77%(with protecting agent).The results suggest that amino acid polymers are good candidates for protein stabilization under extreme conditions.The easy operation of LiHMDS-initiated,moisture insensitive and rapid NCA polymerization implies great potential of this strategy to prepare simple amino acid polymers for screening of protein stabilization agents.
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