Efficient splicing of the CPE intein derived from directed evolution of the <i>Cryptococcus neoformans</i> PRP8 intein.

ABSTRACT: 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 C-extein 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 protei

SUBMITTER: Zhan Q 

PROVIDER: S-EPMC10448054 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature