{"title":"Targeted Peptide Modification Using an Engineered Bacterial N-Glycosyltransferase","authors":"Ayoola B. Smith, Jonathan R. Chekan","doi":"10.1021/acscatal.4c01958","DOIUrl":null,"url":null,"abstract":"Many biosynthetic enzymes involved in the production of ribosomally synthesized and post-translationally modified peptides (RiPPs) in prokaryotic organisms often possess two distinct functional parts: (1) the RiPP precursor peptide recognition element (RRE) and (2) the catalytic domain. The former binds to the conserved leader portion of bipartite precursor peptides, while the latter modifies a specific sequence in the core peptide. This makes the enzymes specific yet promiscuously act on precursor peptides with diverse core sequences. Herein, we engineered fusion enzymes using this biological principle by recombinant tethering of a bacterial <i>N</i>-glycosyltransferase (NGT) to RREs. Also, we designed guided (with a leader peptide) and unguided (no leader peptide) substrates for evaluation of the enzymes. The chimeric system improved the catalytic efficiency as well as the in vitro and in vivo selectivity of the fusion enzyme for some guided substrates compared to the wild-type enzyme. Furthermore, we successfully demonstrated the utility of the engineered enzymes through the production of an <i>N</i>-glycosylated analogue of the glycocin sublancin. Altogether, this work illustrates the viability of our approach for transforming protein post-translational modification enzymes into designer tools for introducing new chemical modifications to RiPP natural products and other peptides.","PeriodicalId":9,"journal":{"name":"ACS Catalysis ","volume":null,"pages":null},"PeriodicalIF":11.3000,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Catalysis ","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1021/acscatal.4c01958","RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, PHYSICAL","Score":null,"Total":0}
引用次数: 0
Abstract
Many biosynthetic enzymes involved in the production of ribosomally synthesized and post-translationally modified peptides (RiPPs) in prokaryotic organisms often possess two distinct functional parts: (1) the RiPP precursor peptide recognition element (RRE) and (2) the catalytic domain. The former binds to the conserved leader portion of bipartite precursor peptides, while the latter modifies a specific sequence in the core peptide. This makes the enzymes specific yet promiscuously act on precursor peptides with diverse core sequences. Herein, we engineered fusion enzymes using this biological principle by recombinant tethering of a bacterial N-glycosyltransferase (NGT) to RREs. Also, we designed guided (with a leader peptide) and unguided (no leader peptide) substrates for evaluation of the enzymes. The chimeric system improved the catalytic efficiency as well as the in vitro and in vivo selectivity of the fusion enzyme for some guided substrates compared to the wild-type enzyme. Furthermore, we successfully demonstrated the utility of the engineered enzymes through the production of an N-glycosylated analogue of the glycocin sublancin. Altogether, this work illustrates the viability of our approach for transforming protein post-translational modification enzymes into designer tools for introducing new chemical modifications to RiPP natural products and other peptides.
期刊介绍:
ACS Catalysis is an esteemed journal that publishes original research in the fields of heterogeneous catalysis, molecular catalysis, and biocatalysis. It offers broad coverage across diverse areas such as life sciences, organometallics and synthesis, photochemistry and electrochemistry, drug discovery and synthesis, materials science, environmental protection, polymer discovery and synthesis, and energy and fuels.
The scope of the journal is to showcase innovative work in various aspects of catalysis. This includes new reactions and novel synthetic approaches utilizing known catalysts, the discovery or modification of new catalysts, elucidation of catalytic mechanisms through cutting-edge investigations, practical enhancements of existing processes, as well as conceptual advances in the field. Contributions to ACS Catalysis can encompass both experimental and theoretical research focused on catalytic molecules, macromolecules, and materials that exhibit catalytic turnover.