Back to Search Start Over

Structural and Functional Analyses of the Tridomain‐Nonribosomal Peptide Synthetase FmoA3 for 4‐Methyloxazoline Ring Formation

Authors :
Kazuo Shin-ya
Yohei Katsuyama
Yasuo Ohnishi
Kaoru Sone
Toshio Moriya
Seiji Kawai
Ayaka Harada
Naoki Urano
Toshiya Senda
Masato Kawasaki
Naruhiko Adachi
Source :
Angewandte Chemie International Edition. 60:14554-14562
Publication Year :
2021
Publisher :
Wiley, 2021.

Abstract

Nonribosomal peptide synthetases (NRPSs) are attractive targets for bioengineering to generate useful peptides . F moA3 is a single modular NRPS composed of heterocyclization (Cy), adenylation (A), and peptidyl carrier protein (PCP) domains. It use s α - methyl- l -serine to synthesize a 4-methyloxazoline ring, probably with another Cy domain in the preceding module FmoA2. Here, we determined the head-to-tail homodimeric structures of FmoA3 by X-ray crystallography ( apo -form, with adenylyl-imidodiphosphate and α -methyl- l -seryl-AMP) and cryogenic electron microscopy single particle analysis, and performed site-directed mutagenesis experiments. The data revealed that α -methyl- l -serine can be accommodated in the active site because of the extra space around Ala688. The Cy domains of FmoA2 and FmoA3 catalyze peptide bond formation and heterocyclization, respectively. FmoA3's Cy domain seems to lose its donor PCP binding activit y. The collective data support a proposed catalytic cycle of FmoA3.

Details

ISSN :
15213773 and 14337851
Volume :
60
Database :
OpenAIRE
Journal :
Angewandte Chemie International Edition
Accession number :
edsair.doi.dedup.....6baf66acaacf62d25832f6872199d2f2
Full Text :
https://doi.org/10.1002/anie.202102760