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

Izumi, Momona; Tomita, Hiroya; Miyazaki, Kentaro; Otsuka, Ryo; Honda, Kohsuke

doi:https://doi.org/10.1002/cbic.202400836

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

アクセス数:536件（2025-08-14 18:37 集計）

固定URL: https://hdl.handle.net/11094/100254

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タイトル	Biosynthetic Characterization of Bacillibactin in Thermophilic Bacillaceae and its Potential for in Vitro Mutational Synthesis
著者	著者名 Izumi, Momona
	NRID 1000000814229 1000000814229 ORCID 0000-0001-7967-921X 0000-0001-7967-921X researchmap permalink htomita htomita 著者名 Tomita, Hiroya
	NRID 1000060344123 1000060344123 ORCID 0000-0002-4349-566X 0000-0002-4349-566X researchmap permalink myzkkntr myzkkntr 著者名 Miyazaki, Kentaro
	著者名 Otsuka, Ryo
	NRID 1000090403162 1000090403162 ORCID 0000-0001-9069-7574 0000-0001-9069-7574 researchmap permalink read0070175 read0070175 Scopus Author ID 7401678792 7401678792 著者名 Honda, Kohsuke
キーワード等	bacillibactin
	biosynthesis
	mutational synthesis
	nonribosomal peptide synthetase
	thermophile
内容	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]
抄録	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.
出版者	John Wiley and Sons Inc
収録物名	ChemBioChem
刊行年月	2025-01-27
言語	英語
ハンドルURL	https://hdl.handle.net/11094/100254
PISSN	14394227
eISSN	14397633
関連情報 (isVersionOf)	DOI (出版社版) https://doi.org/10.1002/cbic.202400836
アクセス権	embargoed access
権利情報	The full-text file will be made open to the public on 27 January 2026 in accordance with the publisher's policy.
出版タイプ	Accepted Manuscript
カテゴリ	学術雑誌論文 Journal Article

資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
DCTERMS.bibliographicCitation	ChemBioChem. 2025
DC.title	Biosynthetic Characterization of Bacillibactin in Thermophilic Bacillaceae and its Potential for in Vitro Mutational Synthesis
DC.creator	Izumi, Momona
	Tomita, Hiroya
	Miyazaki, Kentaro
	Otsuka, Ryo
	Honda, Kohsuke
DC.publisher	John Wiley and Sons Inc
DC.language' scheme='DCTERMS.RFC1766	英語
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2025-01-27
DC.identifier	https://doi.org/10.1002/cbic.202400836
DC.subject	bacillibactin
	biosynthesis
	mutational synthesis
	nonribosomal peptide synthetase
	thermophile
DC.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]
DCTERMS.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.
DC.rights	The full-text file will be made open to the public on 27 January 2026 in accordance with the publisher's policy.
DC.format	100254
citation_title	Biosynthetic Characterization of Bacillibactin in Thermophilic Bacillaceae and its Potential for in Vitro Mutational Synthesis
citation_author	Izumi, Momona
	Tomita, Hiroya
	Miyazaki, Kentaro
	Otsuka, Ryo
	Honda, Kohsuke
citation_publisher	John Wiley and Sons Inc
citation_language	英語
citation_date	2025-01-27
citation_journal_title	ChemBioChem
citation_doi	https://doi.org/10.1002/cbic.202400836
citation_public_url	https://hdl.handle.net/11094/100254

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

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