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2 S-adenosyl-L-methionine + cytidine 5'-{[hydroxy(2-hydroxyethyl)phosphonoyl]phosphate} + reduced acceptor = S-adenosyl-L-homocysteine + 5'-deoxyadenosine + L-methionine + cytidine 5'-{[hydroxy(2-hydroxypropyl)phosphonoyl]phosphate} + oxidized acceptor
2 S-adenosyl-L-methionine + cytidine 5'-{[hydroxy(2-hydroxyethyl)phosphonoyl]phosphate} + reduced acceptor = S-adenosyl-L-homocysteine + 5'-deoxyadenosine + L-methionine + cytidine 5'-{[hydroxy(2-hydroxypropyl)phosphonoyl]phosphate} + oxidized acceptor
proposed mechanism: a 5'-deoxyadenosine radical generated from reductive cleavage of S-adenosyl-L-methionine is used to abstract the hydrogen atom from the C-H bond of the substrate resulting in a substrate radical that reacts with methylcobalamin yielding (S)-2-hydroxypropylphosphonate and cob(II)alamin
2 S-adenosyl-L-methionine + cytidine 5'-{[hydroxy(2-hydroxyethyl)phosphonoyl]phosphate} + reduced acceptor = S-adenosyl-L-homocysteine + 5'-deoxyadenosine + L-methionine + cytidine 5'-{[hydroxy(2-hydroxypropyl)phosphonoyl]phosphate} + oxidized acceptor
proposed mechanism: one electron is transferred from the reduced ironsulfur cluster to S-adenosyl-L-methionine to form an adenosyl radical and methionine. The adenosyl radical abstracts the pro-R hydrogen atom from C2 of (S)-2-hydroxyethylphosphonate, and the resulting substrate radical reacts with methylcobalamin yielding (S)-2-hydroxypropylphosphonate and cob(II)alamin. The enzyme is then returned to the active state by reduction of the 4Fe4S cluster back to the +1 state and binding of S-adenosyl-L-methionine and methylcobalamin. Alternatively, cob(II)alamin might be reduced to cob(I)alamin and methylated while bound to the enzyme
2 S-adenosyl-L-methionine + cytidine 5'-{[hydroxy(2-hydroxyethyl)phosphonoyl]phosphate} + reduced acceptor = S-adenosyl-L-homocysteine + 5'-deoxyadenosine + L-methionine + cytidine 5'-{[hydroxy(2-hydroxypropyl)phosphonoyl]phosphate} + oxidized acceptor
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S-adenosyl-L-methionine:cytidine 5'-{[hydroxy(2-hydroxyethyl)phosphonoyl]phosphate} C-methyltransferase
Requires cobalamin. The enzyme, isolated from the bacterium Streptomyces wedmorensis, is involved in fosfomycin biosynthesis. It is a radical S-adenosyl-L-methionine (SAM) enzyme that contains a [4Fe-4S] center and a methylcob(III)alamin cofactor. The enzyme uses two molecues of SAM for the reaction. One molecule forms a 5'-deoxyadenosyl radical, while the other is used to methylate the cobalamin cofactor. The 5'-deoxyadenosyl radical abstracts a hydrogen from the C2 position of cytidine 5'-{[(2-hydroxyethyl)phosphonoyl]phosphate} forming a free radical that reacts with the methyl group on methylcob(III)alamin at the opposite side from SAM and the [4Fe-4S] cluster to produce a racemic mix of methylated products and cob(II)alamin. Both the [4Fe-4S] cluster and the cob(II)alamin need to be reduced by an unknown factor(s) before the enzyme could catalyse another cycle.
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(2-hydroxyethyl)phosphonate + S-adenosyl-L-methionine + methylcobalamin
(S)-2-hydroxypropylphosphonate + L-methionine + 5'-deoxyadenosine + cob(II)alamin
S-adenosyl-L-methionine + methylcob(III)alamin + 2-hydroxyethylphosphonate
5'-deoxyadenosine + L-methionine + cob(III)alamin + (2S)-2-hydroxypropylphosphonate
(2-hydroxyethyl)phosphonate + S-adenosyl-L-methionine + methylcobalamin
(S)-2-hydroxypropylphosphonate + L-methionine + 5'-deoxyadenosine + cob(II)alamin
Substrates: Fom3 catalyzes the penultimate step in the biosynthesis of fosfomycin, an antibiotic produced by several species of Streptomyces and Pseudomonas
Products: -
?
(2-hydroxyethyl)phosphonate + S-adenosyl-L-methionine + methylcobalamin
(S)-2-hydroxypropylphosphonate + L-methionine + 5'-deoxyadenosine + cob(II)alamin
Substrates: the enzyme is required for fosfomycin production
Products: -
?
(2-hydroxyethyl)phosphonate + S-adenosyl-L-methionine + methylcobalamin
(S)-2-hydroxypropylphosphonate + L-methionine + 5'-deoxyadenosine + cob(II)alamin
Substrates: (S)- and (R)-(2-hydroxy[1-2H1]ethyl)phosphonic acids are fed to Streptomyces fradiae. The deuterium of (R)-(2-hydroxy[1-2H1]ethyl)phosphonic acid is lost, and the deuterium of (S)-(2-hydroxy[1-2H1]ethyl)phosphonic acid is retained on incorporation into fosfomycin
Products: -
?
(2-hydroxyethyl)phosphonate + S-adenosyl-L-methionine + methylcobalamin
(S)-2-hydroxypropylphosphonate + L-methionine + 5'-deoxyadenosine + cob(II)alamin
Substrates: C-2 methylation of hydroxyethyl phosphonate
Products: -
?
(2-hydroxyethyl)phosphonate + S-adenosyl-L-methionine + methylcobalamin
(S)-2-hydroxypropylphosphonate + L-methionine + 5'-deoxyadenosine + cob(II)alamin
Substrates: incorporation of 18O in the alcohol function of 18O-labeled HEP into the FM molecule but not of 18O2 gas
Products: -
?
(2-hydroxyethyl)phosphonate + S-adenosyl-L-methionine + methylcobalamin
(S)-2-hydroxypropylphosphonate + L-methionine + 5'-deoxyadenosine + cob(II)alamin
Substrates: no activity with (R)-2-hydroxyethylphosphonate
Products: -
?
(2-hydroxyethyl)phosphonate + S-adenosyl-L-methionine + methylcobalamin
(S)-2-hydroxypropylphosphonate + L-methionine + 5'-deoxyadenosine + cob(II)alamin
Substrates: when (R)- and (S)-2-hydroxyethylphosphonate are fed to Streptomyces fradiae producing fosfomycin, only (S)-2-hydroxyethylphosphonate is efficiently incorporated into fosfomycin (37% deuterium at C-1 of the aminophosphonic acid)
Products: -
?
(2-hydroxyethyl)phosphonate + S-adenosyl-L-methionine + methylcobalamin
(S)-2-hydroxypropylphosphonate + L-methionine + 5'-deoxyadenosine + cob(II)alamin
Substrates: -
Products: -
?
S-adenosyl-L-methionine + methylcob(III)alamin + 2-hydroxyethylphosphonate
5'-deoxyadenosine + L-methionine + cob(III)alamin + (2S)-2-hydroxypropylphosphonate
Substrates: -
Products: -
?
S-adenosyl-L-methionine + methylcob(III)alamin + 2-hydroxyethylphosphonate
5'-deoxyadenosine + L-methionine + cob(III)alamin + (2S)-2-hydroxypropylphosphonate
Substrates: the enzyme is involved in the biosynthesis of the broad-spectrum antibiotic fosfomycin
Products: -
?
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(2-hydroxyethyl)phosphonate + S-adenosyl-L-methionine + methylcobalamin
(S)-2-hydroxypropylphosphonate + L-methionine + 5'-deoxyadenosine + cob(II)alamin
S-adenosyl-L-methionine + methylcob(III)alamin + 2-hydroxyethylphosphonate
5'-deoxyadenosine + L-methionine + cob(III)alamin + (2S)-2-hydroxypropylphosphonate
Substrates: the enzyme is involved in the biosynthesis of the broad-spectrum antibiotic fosfomycin
Products: -
?
(2-hydroxyethyl)phosphonate + S-adenosyl-L-methionine + methylcobalamin
(S)-2-hydroxypropylphosphonate + L-methionine + 5'-deoxyadenosine + cob(II)alamin
Substrates: Fom3 catalyzes the penultimate step in the biosynthesis of fosfomycin, an antibiotic produced by several species of Streptomyces and Pseudomonas
Products: -
?
(2-hydroxyethyl)phosphonate + S-adenosyl-L-methionine + methylcobalamin
(S)-2-hydroxypropylphosphonate + L-methionine + 5'-deoxyadenosine + cob(II)alamin
Substrates: the enzyme is required for fosfomycin production
Products: -
?
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methylcobalamin
a B12-derived cofactor. A conserved domain search of the Fom3 sequence shows that it has two conserved domains. The N-terminal domain is identified as a B12-like binding domain, whereas the C-terminal domain shows homology to the radical-SAM protein family, containing three conserved Cys residues that serve as ligands to a [4Fe-4S] cluster
methylcobalamin
proposed mechanism: a 5'-deoxyadenosine radical generated from reductive cleavage of S-adenosyl-L-methionine is used to abstract the hydrogen atom from the C-H bond of the substrate resulting in a substrate radical that reacts with methylcobalamin yielding (S)-2-hydroxypropylphosphonate and cob(II)alamin
S-adenosyl-L-methionine
a conserved domain search of the Fom3 sequence shows it has two conserved domains. The N-terminal domain is identified as a B12-like binding domain, whereas the C-terminal domain shows homology to the radical-SAM protein family, containing three conserved Cys residues that serve as ligands to a [4Fe-4S] cluster
S-adenosyl-L-methionine
proposed mechanism: a 5'-deoxyadenosine radical generated from reductive cleavage of S-adenosyl-L-methionine is used to abstract the hydrogen atom from the C-H bond of the substrate resulting in a substrate radical that reacts with methylcobalamin yielding (S)-2-hydroxypropylphosphonate and cob(II)alamin
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C282A
the mutant enzyme is not able to produce the catalytic [4Fe-4S]+1 cluster
C282A/C286A/C289A
the triple-variant is very unstable and precipitated during purification and subsequent manipulation for experiments. It completely lacks the [4Fe-4S]+1 cluster
C286A
the mutant enzyme is not able to produce the catalytic [4Fe-4S]+1 cluster
C289A
the mutant enzyme is not able to produce the catalytic [4Fe-4S]+1 cluster
additional information
a mutant of Streptomyces wedmorensis (A16), defective in the biosynthesis of cobalamin produces fosfomycin only in the presence of hydroxocobalamin. Incorporation of [14CH3]methylcobalamin into fosfomycin by the mutant A16
additional information
a mutant of Streptomyces wedmorensis (A16), defective in the biosynthesis of cobalamin produces fosfomycin only in the presence of hydroxocobalamin. Incorporation of [14CH3]methylcobalamin into fosfomycin by the mutant A16
additional information
site-directed mutagenesis of the cysteine residues in the radical SAM CxxxCxxC motif indicates that each residue is essential for functional cluster formation
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Woodyer, R.D.; Shao, Z.; Thomas, P.M.; Kelleher, N.L.; Blodgett, J.A.; Metcalf, W.W.; van der Donk, W.A.; Zhao, H.
Heterologous production of fosfomycin and identification of the minimal biosynthetic gene cluster
Chem. Biol.
13
1171-1182
2006
Streptomyces fradiae (D2SNF5)
brenda
Woodyer, R.D.; Li, G.; Zhao, H.; van der Donk, W.A.
New insight into the mechanism of methyl transfer during the biosynthesis of fosfomycin
Chem. Commun. (Camb. )
4
359-361
2007
Streptomyces fradiae (D2SNF5)
brenda
Booker, S.J.
Anaerobic functionalization of unactivated C-H bonds
Curr. Opin. Chem. Biol.
13
58-73
2009
Streptomyces fradiae (D2SNF5)
brenda
Kuzuyama, T.; Hidaka, T.; Kamigiri, K.; Imai, S.; Seto, H.
Studies on the biosynthesis of fosfomycin. 4. The biosynthetic origin of the methyl group of fosfomycin
J. Antibiot.
45
1812-1814
1992
Streptomyces wedmorensis (Q56184)
brenda
Hammerschmidt, F.
Biosynthesis of natural products with a P-C bond. Part 8: on the origin of the oxirane oxygen atom of fosfomycin in Streptomyces fradiae
J. Chem. Soc. Perkin Trans.
1
1993-1996
1991
Streptomyces fradiae (D2SNF5)
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brenda
Hammerschmidt, F.; Khlig, H.
Biosynthesis of natural products with a P-C Bond. 7. Synthesis of [1,1-2H2]-, [2,2-2H2]-, (R)- and (S)-[2 H1](2-hydroxyethyl)phosphonic acid and (R,S)-[1-2H1](1,2-dihydroxyethyl)phosphonic acid and incorporation studies into fosfomycin in Streptomyces fradiae
J. Org. Chem.
56
2364-2370
1991
Streptomyces fradiae (D2SNF5)
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brenda
Hidaka, T.; Goda, M.; Kuzuyama, T.; Takei, N.; Hidaka, M.; Seto, H.
Cloning and nucleotide sequence of fosfomycin biosynthetic genes of Streptomyces wedmorensis
Mol. Gen. Genet.
249
274-280
1995
Streptomyces wedmorensis (Q56184), Streptomyces wedmorensis 144-91 (Q56184)
brenda
Seto, H.; Kuzuyama, T.
Bioactive natural products with carbon-phosphorus bonds and their biosynthesis
Nat. Prod. Rep.
16
589-596
1999
Streptomyces fradiae (D2SNF5), Streptomyces wedmorensis (Q56184)
brenda
Allen, K.D.; Wang, S.C.
Initial characterization of Fom3 from Streptomyces wedmorensis: The methyltransferase in fosfomycin biosynthesis
Arch. Biochem. Biophys.
543
67-73
2014
Streptomyces wedmorensis (Q56184)
brenda