An Enzymatic Pathway for the Biosynthesis of the Formylhydroxyornithine Required for Rhodochelin Iron Coordination

Bosello, Mattia; Mielcarek, Andreas; Giessen, Tobias W.; Marahiel, Mohamed A.

doi:10.1021/bi201837f

Cited by 31 publications

(39 citation statements)

References 34 publications

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“…12−23 Here we investigate the l -Orn-specific N -monooxygenase from Kutzneria sp. 744, KtzI, which instead provides a building block for the biosynthesis of a new class of antifungal antimicrobials called kutznerides 7 (Figure 1a).…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Crystallographic Evidence of Drastic Conformational Changes in the Active Site of a Flavin-Dependent N-Hydroxylase

Setser¹,

Heemstra

Walsh

et al. 2014

Biochemistry

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The soil actinomycete Kutzneria sp. 744 produces a class of highly decorated hexadepsipeptides, which represent a new chemical scaffold that has both antimicrobial and antifungal properties. These natural products, known as kutznerides, are created via nonribosomal peptide synthesis using various derivatized amino acids. The piperazic acid moiety contained in the kutzneride scaffold, which is vital for its antibiotic activity, has been shown to derive from the hydroxylated product of l-ornithine, l-N5-hydroxyornithine. The production of this hydroxylated species is catalyzed by the action of an FAD- and NAD(P)H-dependent N-hydroxylase known as KtzI. We have been able to structurally characterize KtzI in several states along its catalytic trajectory, and by pairing these snapshots with the biochemical and structural data already available for this enzyme class, we propose a structurally based reaction mechanism that includes novel conformational changes of both the protein backbone and the flavin cofactor. Further, we were able to recapitulate these conformational changes in the protein crystal, displaying their chemical competence. Our series of structures, with corroborating biochemical and spectroscopic data collected by us and others, affords mechanistic insight into this relatively new class of flavin-dependent hydroxylases and adds another layer to the complexity of flavoenzymes.

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Section: Discussionmentioning

confidence: 99%

“…7 This specificity for the substrate, and the cofactor and cosubstrate usage, is similar to that of previously characterized flavin N -hydroxylases. 12−23 However, KtzI does differ from these other systems in the fate of its hydroxylated product.…”

mentioning

confidence: 99%

Crystallographic Evidence of Drastic Conformational Changes in the Active Site of a Flavin-Dependent N-Hydroxylase

Setser¹,

Heemstra

Walsh

et al. 2014

Biochemistry

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“…genomes. This siderophore BGC was composed of seven open reading frames (ORFs), which were annotated with functions consistent with iron acquisition and siderophore biosynthesis (70–72) ( Fig. 4A ).…”

Section: Resultsmentioning

confidence: 99%

Bacterial competition mediated by siderophore production among the human nasal microbiota

Stubbendieck

May

Chevrette

et al. 2018

Preprint

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19Resources available in the human nasal cavity are limited. Therefore, to successfully 20 colonize the nasal cavity, bacteria must compete for scarce nutrients. Competition may 21 occur directly through interference (e.g., antibiotics) or indirectly by nutrient 22 sequestration. To investigate the nature of nasal bacterial competition, we performed 23 co-culture inhibition assays between nasal Actinobacteria and Staphylococcus spp. We 24 found that Staphylococcus epidermidis isolates were sensitive to growth inhibition by 25 Actinobacteria but Staphylococcus aureus isolates were resistant to inhibition. Among 26 Actinobacteria, we observed that Corynebacterium spp. were variable in their ability to 27 inhibit S. epidermidis. We sequenced the genomes of ten Corynebacterium spp. 28 isolates, including three Corynebacterium propinquum that strongly inhibited S. 29 epidermidis and seven other Corynebacterium spp. isolates that only weakly inhibited S. 30 epidermidis. Using a comparative genomics approach, we found that the C. propinquum 31 genomes were enriched in genes for iron acquisition and encoded a biosynthetic gene 32 cluster (BGC) for siderophore production, absent in the non-inhibitory Corynebacterium 33 spp. genomes. Using a chromeazurol S assay, we confirmed that C. propinquum 34 produced siderophores. We demonstrated that iron supplementation rescued S. 35 epidermidis from inhibition by C. propinquum, suggesting that inhibition was due to iron 36 restriction through siderophore production. Using comparative metabolomics, we 37 identified the siderophore produced by C. propinquum as dehydroxynocardamine. 38

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“…In 2011, two structures of this enzyme, one in its oxidized state and the other in its reduced state, were presented for the first time (Olucha et al, 2011). An L-ornithine hydroxylase was identified in the proteome of R. jostii RHA1 (Bosello et al, 2012) and, its coding sequence was cloned and expressed from a pBAD-based expression vector just recently (Riebel et al, 2013a). …”

Section: N-hydroxylations and N-oxidationsmentioning

confidence: 99%

Flavoprotein monooxygenases for oxidative biocatalysis: recombinant expression in microbial hosts and applications

2014

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External flavoprotein monooxygenases comprise a group of flavin-dependent oxidoreductases that catalyze the insertion of one atom of molecular oxygen into an organic substrate and the second atom is reduced to water. These enzymes are involved in a great number of metabolic pathways both in prokaryotes and eukaryotes. Flavoprotein monooxygenases have attracted the attention of researchers for several decades and the advent of recombinant DNA technology caused a great progress in the field. These enzymes are subjected to detailed biochemical and structural characterization and some of them are also regarded as appealing oxidative biocatalysts for the production of fine chemicals and valuable intermediates toward active pharmaceutical ingredients due to their high chemo-, stereo-, and regioselectivity. Here, we review the most representative reactions catalyzed both in vivo and in vitro by prototype flavoprotein monooxygenases, highlighting the strategies employed to produce them recombinantly, to enhance the yield of soluble proteins, and to improve cofactor regeneration in order to obtain versatile biocatalysts. Although we describe the most outstanding features of flavoprotein monooxygenases, we mainly focus on enzymes that were cloned, expressed and used for biocatalysis during the last years.

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An Enzymatic Pathway for the Biosynthesis of the Formylhydroxyornithine Required for Rhodochelin Iron Coordination

Cited by 31 publications

References 34 publications

Crystallographic Evidence of Drastic Conformational Changes in the Active Site of a Flavin-Dependent N-Hydroxylase

Crystallographic Evidence of Drastic Conformational Changes in the Active Site of a Flavin-Dependent N-Hydroxylase

Bacterial competition mediated by siderophore production among the human nasal microbiota

Flavoprotein monooxygenases for oxidative biocatalysis: recombinant expression in microbial hosts and applications

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