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Information on EC 1.14.99.65 - 4-amino-L-phenylalanyl-[CmlP-peptidyl-carrier-protein] 3-hydroxylase and Organism(s) Streptomyces venezuelae and UniProt Accession F2RB80

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IUBMB Comments
The enzyme, characterized from the bacterium Streptomyces venezuelae, participates in the biosynthesis of the antibiotic chloramphenicol. It carries an oxygen-bridged dinuclear iron cluster. The native electron donor remains unknown, and the enzyme was assayed in vitro using sodium dithionite. The enzyme only acts on its substrate when it is loaded onto the peptidyl-carrier domain of the CmlP non-ribosomal peptide synthase.
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Streptomyces venezuelae
UNIPROT: F2RB80
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The taxonomic range for the selected organisms is: Streptomyces venezuelae
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Reaction Schemes
4-amino-L-phenylalanyl-[CmlP-peptidyl-carrier-protein]
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2-(4-aminophenyl)-L-seryl-[CmlP-peptidyl-carrier-protein]
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Synonyms
cmlA, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
cmlA
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PATHWAY SOURCE
PATHWAYS
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SYSTEMATIC NAME
IUBMB Comments
4-amino-L-phenylalanyl-[CmlP-peptidyl-carrier-protein],acceptor:oxygen 3-oxidoreductase
The enzyme, characterized from the bacterium Streptomyces venezuelae, participates in the biosynthesis of the antibiotic chloramphenicol. It carries an oxygen-bridged dinuclear iron cluster. The native electron donor remains unknown, and the enzyme was assayed in vitro using sodium dithionite. The enzyme only acts on its substrate when it is loaded onto the peptidyl-carrier domain of the CmlP non-ribosomal peptide synthase.
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-amino-L-phenylalanyl-[CmlP-peptidyl-carrier-protein] + reduced acceptor + O2
2-(4-aminophenyl)-L-seryl-[CmlP-peptidyl-carrier-protein] + acceptor + H2O
show the reaction diagram
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additional information
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CmlA contains an oxo-bridged dinuclear iron cluster. CmlA is functional in the diferrous state, and substrate L-PAPA is covalently bound to CmlP. The product is hydroxylated L-PAPA and O2 is the oxygen source
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
structure in the diferrous state. Formation of the active diferrous state CmlA-CmlP-L-p-aminophenylalanine complex converts at least one iron of the clusters from six- to five-coordinate by changing a bidentately bound amino acid carboxylate to monodentate on Fe1. The only bidentate carboxylate is residue E377
to 2.7 A resolution. The C-terminal domain has the alphabetabetaalpha fold of the metallo-beta-lactamase superfamily. The N-terminal domain facilitates dimerization, and likely serves additional roles in nonribosomal peptide synthetase recognition and the regulation of O2 activation
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E377D
variant E377D shows only monodentate carboxylate coordination. Reduced E377D reacts rapidly with O2, showing loss of regulation. The mutant fails to catalyze hydroxylation
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Makris, T.M.; Knoot, C.J.; Wilmot, C.M.; Lipscomb, J.D.
Structure of a dinuclear iron cluster-containing beta-hydroxylase active in antibiotic biosynthesis
Biochemistry
52
6662-6671
2013
Streptomyces venezuelae (F2RB80), Streptomyces venezuelae DSM 40230 (F2RB80)
Manually annotated by BRENDA team
Jasniewski, A.J.; Knoot, C.J.; Lipscomb, J.D.; Que, L.
A carboxylate shift regulates dioxygen activation by the diiron nonheme beta-hydroxylase CmlA upon binding of a substrate-loaded nonribosomal peptide synthetase
Biochemistry
55
5818-5831
2016
Streptomyces venezuelae (F2RB80), Streptomyces venezuelae DSM 40230 (F2RB80)
Manually annotated by BRENDA team
Vu, V.V.; Makris, T.M.; Lipscomb, J.D.; Que, L.
Active-site structure of a beta-hydroxylase in antibiotic biosynthesis
J. Am. Chem. Soc.
133
6938-6941
2011
Streptomyces venezuelae (F2RB80), Streptomyces venezuelae DSM 40230 (F2RB80)
Manually annotated by BRENDA team
Makris, T.M.; Chakrabarti, M.; Muenck, E.; Lipscomb, J.D.
A family of diiron monooxygenases catalyzing amino acid beta-hydroxylation in antibiotic biosynthesis
Proc. Natl. Acad. Sci. USA
107
15391-15396
2010
Streptomyces venezuelae (F2RB80), Streptomyces venezuelae DSM 40230 (F2RB80)
Manually annotated by BRENDA team