Functional Identification of Putrescine C- and N-Hydroxylases

Bin Li, Tiffany Lowe-Power, Shin Kurihara, Stephen Gonzales, Jacinth Naidoo, John B. Macmillan, Caitilyn Allen, Anthony J. Michael

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The small polyamine putrescine (1,4-diaminobutane) is ubiquitously and abundantly found in all three domains of life. It is a precursor, through N-aminopropylation or N-aminobutylation, for biosynthesis of the longer polyamines spermidine, sym-homospermidine, spermine, and thermospermine and longer and branched chain polyamines. Putrescine is also biochemically modified for purposes of metabolic regulation and catabolism, e.g. N-acetylation and N-glutamylation, and for incorporation into specialized metabolites, e.g. N-methylation, N-citrylation, N-palmitoylation, N-hydroxylation, and N-hydroxycinnamoylation. Only one example is known where putrescine is modified on a methylene carbon: the formation of 2-hydroxyputrescine by an unknown C-hydroxylase. Here, we report the functional identification of a previously undescribed putrescine 2-hydroxylase, a Rieske-type nonheme iron sulfur protein from the β-proteobacteria Bordetella bronchiseptica and Ralstonia solanacearum. Identification of the putrescine 2-hydroxylase will facilitate investigation of the physiological functions of 2-hydroxyputrescine. One known role of 2-hydroxyputrescine has direct biomedical relevance: its role in the biosynthesis of the cyclic hydroxamate siderophore alcaligin, a potential virulence factor of the causative agent of whooping cough, Bordetella pertussis. We also report the functional identification of a putrescine N-hydroxylase from the γ-proteobacterium Shewanella oneidensis, which is homologous to FAD- and NADPH-dependent ornithine and lysine N-monooxygenases involved in siderophore biosynthesis. Heterologous expression of the putrescine N-hydroxylase in E. coli produced free N-hydroxyputrescine, never detected previously in a biological system. Furthermore, the putrescine C- and N-hydroxylases identified here could contribute new functionality to polyamine structural scaffolds, including C-H bond functionalization in synthetic biology strategies.

Original languageEnglish (US)
Pages (from-to)2782-2789
Number of pages8
JournalACS Chemical Biology
Volume11
Issue number10
DOIs
StatePublished - Oct 21 2016

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Putrescine
Mixed Function Oxygenases
Polyamines
Biosynthesis
Siderophores
Proteobacteria
Nonheme Iron Proteins
Ralstonia solanacearum
Bordetella bronchiseptica
Shewanella
Lipoylation
Iron-Sulfur Proteins
Synthetic Biology
Acetylation
Bordetella pertussis
Hydroxylation
Flavin-Adenine Dinucleotide
Ornithine
Methylation
Spermidine

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

Functional Identification of Putrescine C- and N-Hydroxylases. / Li, Bin; Lowe-Power, Tiffany; Kurihara, Shin; Gonzales, Stephen; Naidoo, Jacinth; Macmillan, John B.; Allen, Caitilyn; Michael, Anthony J.

In: ACS Chemical Biology, Vol. 11, No. 10, 21.10.2016, p. 2782-2789.

Research output: Contribution to journalArticle

Li, B, Lowe-Power, T, Kurihara, S, Gonzales, S, Naidoo, J, Macmillan, JB, Allen, C & Michael, AJ 2016, 'Functional Identification of Putrescine C- and N-Hydroxylases', ACS Chemical Biology, vol. 11, no. 10, pp. 2782-2789. https://doi.org/10.1021/acschembio.6b00629
Li B, Lowe-Power T, Kurihara S, Gonzales S, Naidoo J, Macmillan JB et al. Functional Identification of Putrescine C- and N-Hydroxylases. ACS Chemical Biology. 2016 Oct 21;11(10):2782-2789. https://doi.org/10.1021/acschembio.6b00629
Li, Bin ; Lowe-Power, Tiffany ; Kurihara, Shin ; Gonzales, Stephen ; Naidoo, Jacinth ; Macmillan, John B. ; Allen, Caitilyn ; Michael, Anthony J. / Functional Identification of Putrescine C- and N-Hydroxylases. In: ACS Chemical Biology. 2016 ; Vol. 11, No. 10. pp. 2782-2789.
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