Work

Investigation of Amide Bond Formation during Dehydrophos Biosynthesis

Year: 2018

Type: article

Abstract: Dehydrophos is a tripeptide phosphonate antibiotic produced by Streptomyces luridus . Its biosynthetic pathway involves the use of aminoacyl-tRNA (aa-tRNA) for amide bond formation. The first amide bond during biosynthesis is formed by DhpH-C, a peptidyltransferase that utilizes Leu-tRNA Leu . DhpH-C is a member of a burgeoning family of natural product biosynthetic enzymes that make use of aa-tRNA outside of canonical translation activities in the cell. Here, we used site-directed mutagenesis of both DhpH-C and tRNA Leu to investigate the enzyme mechanism and substrate specificity, respectively, and analyzed the substrate scope for the production of a set of dipeptides. DhpH-C appears to recognize both the amino acyl group on the tRNA and the tRNA acceptor stem, and the enzyme can accept other hydrophobic residues, in addition to leucine. These results contribute to a better understanding of enzyme-aa-tRNA interactions and the growing exploration of aa-tRNA usage beyond translation.

Source: ACS Chemical Biology

DOI: 10.1021/acschembio.7b00949

Authors: Emily C. Ulrich, Despina J. Bougioukou, Wilfred A. van der Donk

Institutions: University of Illinois Urbana-Champaign, Carl R. Woese Institute for Genomic Biology, Howard Hughes Medical Institute

Language: English

Topic: Microbial Natural Products and Biosynthesis

Subfield: Applied Microbiology and Biotechnology

Field: Immunology and Microbiology

Domain: Life Sciences

Open Access status: green

Funders: National Institutes of Health, National Institute of General Medical Sciences, National Center for Research Resources, Carl R. Woese Institute for Genomic Biology

Awards: G1554975086, Discovery, Design, and Development of Phosphonic Acid Antibiotics, Request for a Varian 600 MHz NMR Instrument