Efficient splicing of the CPE intein derived from directed evolution of the Cryptococcus neoformans PRP8 intein  

作　　者：Qin Zhan Changhua Shi Yu Jiang Xianling Gao Ying Lin 

机构地区：[1]College of Biological Science and Medical Engineering,Donghua University,Shanghai 201620,China [2]Shandong Guoli Biotechnology Co.,Ltd.,Jinan 250101,China

出　　处：《Acta Biochimica et Biophysica Sinica》2023年第8期1310-1318,共9页生物化学与生物物理学报（英文版）

基　　金：supported by the grants from the Science and Technology Commission of Shanghai Municipality(No.19ZR1471000);the National Natural Science Foundation of China(No.31470836).

摘　　要：Intein-mediated protein splicing has been widely used in protein engineering;however,the splicing efficiency and extein specificity usually limit its further application.Thus,there is a demand for more general inteins that can overcome these limitations.Here,we study the trans-splicing of CPE intein obtained from the directed evolution of Cne PRP8,which shows that its splicing rate is~29-fold higher than that of the wild-type.When the+1 residue of Cextein is changed to cysteine,CPE also shows high splicing activity.Faster association and higher affinity may contribute to the high splicing rate compared with wild-type intein.These findings have important implications for the future engineering of inteins and provide clues for fundamental studies of protein structure and folding.

关 键 词：Cne PRP8 intein C-extein directed evolution 

分 类 号：R563.1[医药卫生—呼吸系统]