Peptide synthesis cooperatively achieved by peptide ligase and ribosomes

2014 Fiscal Year Final Research Report

• Project/Area Number: 25560397
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biomolecular chemistry
• Research Institution: Hokkaido University
• Principal Investigator: DAIRI Tohru 北海道大学, 工学(系)研究科(研究院), 教授 (70264679)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: フェガノマイシン / ペプチド抗生物質 / 放線菌 / ペプチドライゲース

Outline of Final Research Achievements

Pheganomycin (PGM) (1) consists of a nonproteinogenic amino acid, (S)-2-(3,5-dihydroxy-4-methoxyphenyl)-2-guanidinoacetic acid (2) at the N-terminus and a proteinogenic core peptide derived from NVKDGPT or NVKDR. The biosynthetic gene cluster was identified in Streptomyces cirratus to contain a gene encoding a precursor peptide, which included both the core peptides, and several genes plausibly encoding enzymes for 2 biosynthesis. We identified a gene (pgm1) responsible for the peptide bond formation between 2 and the peptides in the cluster. A pgm1-disruptant lost 1 productivity and recombinant PGM1 catalyzed the ATP-dependent peptide bond formation. This is the first example of cooperative peptide synthesis achieved by ribosome and peptide ligase using a peptide as a nucleophile. PGM1 accepted a variety of peptides as the nucleophile and the flexibility was comprehended by the crystal structure of PGM1 and the mutagenesis analyses.

Free Research Field

生合成工学