Substrate selection of adenylation domains for nonribosomal peptide synthetase (NRPS) in bacillamide C biosynthesis by marine Bacillus atrophaeus C89

• Published in: Journal of industrial microbiology & biotechnology Vol. 45; no. 5; pp. 335 - 344
• Main Authors: Zhang, Fengli, Wang, Yukun, Jiang, Qun, Chen, Qihua, Karthik, Loganathan, Zhao, Yi-Lei, Li, Zhiyong
• Format: Journal Article
• Language: English
• Published: Cham Springer International Publishing 01-05-2018; Oxford University Press
• Subjects: Adenylation; Alanine; Bacillus - genetics; Bacillus - metabolism; Bacillus atrophaeus; Biochemistry; Bioinformatics; Biomedical and Life Sciences; Biosynthesis; Biotechnology; Biotechnology Methods - Original Paper; Clusters; Crystal structure; Cysteine; Cysteine - metabolism; Genetic Engineering; Genomes; Homology; Inorganic Chemistry; Life Sciences; Microbiology; Molecular docking; Molecular Docking Simulation; Natural products; Peptide Synthases - metabolism; Peptides; Protein Domains; Proteins; Structural analysis; Substrate Specificity; Substrates; Thiazoles - metabolism; Tryptamines - metabolism; Nonribosomal peptide synthetases (NRPS); Adenylation domain; Substrates selection; Bacillamide C; Bacillus atrophaeus
• ISSN: 1367-5435; 1476-5535
• DOI: 10.1007/s10295-018-2028-2

Nonribosomal peptide synthetases (NRPSs) are multi-modular enzymes involved in the biosynthesis of natural products. Bacillamide C was synthesized by
Bacillus atrophaeus
C89. A nonribosomal peptide synthetase (NRPS) cluster found in the genome of
B. atrophaeus
C89 was hypothesized to be responsible for the biosynthesis of bacillamide C using alanine and cysteine as substrates. Here, the structure analysis of adenylation domains based on homologous proteins with known crystal structures indicated locations of the substrate-binding pockets. Molecular docking suggested alanine and cysteine as the potential substrates for the two adenylation domains in the NRPS cluster. Furthermore, biochemical characterization of the purified recombinant adenylation domains proved that alanine and cysteine were the optimum substrates for the two adenylation domains. The results provided the in vitro evidence for the hypothesis that the two adenylation domains in the NRPS of
B. atrophaeus
C89 preferentially select alanine and cysteine, respectively, as a substrate to synthesize bacillamide C. Furthermore, this study on substrates selectivity of adenylation domains provided basis for rational design of bacillamide analogs.