Biosynthetic Characterization of Bacillibactin in Thermophilic Bacillaceae and its Potential for in Vitro Mutational Synthesis

Izumi, Momona; Tomita, Hiroya; Miyazaki, Kentaro et al.

ChemBioChem, 2025

Number of Access:555（2025-09-17 08:09 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/100254

Link to primary information 

File Format Terms of use Size Views Date.Available Description information
ChemBioChem_cbic.202400836 pdf None 5.98 MB   2026.01.27  

Item Information

Output File Export EndNote Basic Export Mendeley

Title
Biosynthetic Characterization of Bacillibactin in Thermophilic Bacillaceae and its Potential for in Vitro Mutational Synthesis
Creator
CreatorName
Izumi, Momona
NRID
1000000814229 1000000814229
ORCID
0000-0001-7967-921X 0000-0001-7967-921X
researchmap permalink
htomita htomita
CreatorName
Tomita, Hiroya
NRID
1000060344123 1000060344123
ORCID
0000-0002-4349-566X 0000-0002-4349-566X
researchmap permalink
myzkkntr myzkkntr
CreatorName
Miyazaki, Kentaro
CreatorName
Otsuka, Ryo
NRID
1000090403162 1000090403162
ORCID
0000-0001-9069-7574 0000-0001-9069-7574
researchmap permalink
read0070175 read0070175
Scopus Author ID
7401678792 7401678792
CreatorName
Honda, Kohsuke
Subject
bacillibactin
biosynthesis
mutational synthesis
nonribosomal peptide synthetase
thermophile
Description
This is the accepted version of the following article: Izumi M., Tomita H., Miyazaki K., et al. Biosynthetic Characterization of Bacillibactin in Thermophilic Bacillaceae and its Potential for in Vitro Mutational Synthesis. ChemBioChem, (2025), which has been published in final form at https://doi.org/10.1002/cbic.202400836. This article may be used for non-commercialpurposes in accordance with the Wiley Self-ArchivingPolicy [https://authorservices.wiley.com/author-resources/Journal-Authors/licensing/self-archiving.html]
Abstract
Bacillibactin (BB) is a microbial siderophore produced by Bacillus species. BB is biosynthesized from 2,3-dihydroxybenzoic acid (2,3-DHB), Gly, and L-Thr by nonribosomal peptide synthetase (NRPS) enzymes DhbE, DhbB, and DhbF. The biosynthetic gene cluster (dhb) is also conserved in some strains of thermophilic genera, Geobacillus, Anoxybacillus and Parageobacillus. However, the production of BB from these thermophilic bacteria has not been characterized. Here, we report in vivo and in vitro characterization of BB biosynthesis in Parageobacillus sp. KH3-4 which grows at 65 °C. We confirmed BB production in this thermophilic bacterium and the gene cluster active. In vitro enzymatic analysis revealed that 4′-phosphopantetheinyltransferase (PPTase) encoded in the same gene cluster is responsible for the post-translational maturation of carrier proteins. DhbE and DhbF showed substrate preference to 2,3-DHB and Gly and L-Thr, respectively, consistent with the chemical structure of BB. With the purified enzymes, we successfully reconstituted the NRPS assembly line in vitro. In addition, using chemically synthesized acyl-N-acetylcysteamine substrate analogues, BB analogues possessing methylbenzoyl groups instead of 2,3-DHB were detected. This study provides a new insight into secondary metabolism in thermophiles, and it expands the temperature limitation of NRPS enzymes.
Publisher
John Wiley and Sons Inc
Source Title
ChemBioChem
Date of Issued
2025-01-27
Language
English
Handle URL
https://hdl.handle.net/11094/100254
PISSN
14394227
eISSN
14397633
Relation.isVersionOf
DOI (Publisher Version)
https://doi.org/10.1002/cbic.202400836
Access Rights
embargoed access
CopyRight
The full-text file will be made open to the public on 27 January 2026 in accordance with the publisher's policy.
oaire:version
Accepted Manuscript
Category
学術雑誌論文 Journal Article