Information on EC 2.1.1.326 - N-acetyldemethylphosphinothricin P-methyltransferase

BcpD

P-methylase

PhpK

SD_1168

2 S-adenosyl-L-methionine + N-acetyldemethylphosphinothricin + reduced acceptor = S-adenosyl-L-homocysteine + 5'-deoxyadenosine + L-methionine + N-acetylphosphinothricin + oxidized acceptor

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KEGG

Biosynthesis of secondary metabolites, Phosphonate and phosphinate metabolism

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Biosynthesis of secondary metabolites, Biosynthesis of secondary metabolites

S-adenosyl-L-methionine:N-acetyldemethylphosphinothricin P-methyltransferase

The enzyme was originally characterized from bacteria that produce the tripeptides bialaphos and phosalacine, which inhibit plant and bacterial glutamine synthetases. It is a radical S-adenosyl-L-methionine (SAM) enzyme that contains a [4Fe-4S] center and a methylcob(III)alamin cofactor. According to the proposed mechanism, the reduced iron-sulfur center donates an electron to SAM, resulting in homolytic cleavage of the carbon-sulfur bond to form a 5'-deoxyadenosyl radical that abstracts the hydrogen atom from the P-H bond of the substrate, forming a phosphinate-centered radical. This radical reacts with methylcob(III)alamin to produce the methylated product and cob(II)alamin, which is reduced by an unknown donor to cob(I)alamin. A potential route for restoring the latter back to methylcob(III)alamin is a nucleophilic attack on a second SAM molecule. The enzyme acts in vivo on N-acetyldemethylphosphinothricin-L-alanyl-L-alanine or N-acetyl-demethylphosphinothricin-L-alanyl-L-leucine, the intermediates in the biosynthesis of bialaphos and phosalacine, respectively. This transformation produces the only example of a carbon-phosphorus-carbon linkage known to occur in nature.

2 S-adenosyl-L-methionine + demethylphosphinothricin + reduced acceptor

S-adenosyl-L-homocysteine + 5'-deoxyadenosine + L-methionine + phosphinothricin + oxidized acceptor

2 entries

2 S-adenosyl-L-methionine + N-acetyldemethylphosphinothricin + reduced acceptor

S-adenosyl-L-homocysteine + 5'-deoxyadenosine + L-methionine + N-acetylphosphinothricin + oxidized acceptor

6 entries

S-adenosyl-L-methionine + N-acetyldemethyl phosphinothricin tripeptide + reduced acceptor

S-adenosyl-L-homocysteine + 5'-deoxyadenosine + L-methionine + N-acetylphosphinothricin tripeptide + oxidized acceptor

2 entries

S-adenosyl-L-methionine + N-acetyldemethylbialaphos + reduced acceptor

?

2 entries

S-adenosyl-L-methionine + N-acetyldemethylphosphinothricin + reduced acceptor

S-adenosyl-L-homocysteine + 5'-deoxyadenosine + L-methionine + N-acetylphosphinothricin + oxidized acceptor

2 entries

2 S-adenosyl-L-methionine + N-acetyldemethylphosphinothricin + reduced acceptor

S-adenosyl-L-homocysteine + 5'-deoxyadenosine + L-methionine + N-acetylphosphinothricin + oxidized acceptor

Kitasatospora phosalacinea

A0A059U454

N-acetyldemethylphosphinothricin i.e. (2S)-2-acetamido-4-phosphinatobutanoate. In vivo the enzyme acts on N-acetyldemethylphosphinothricin-L-alanyl-L-alanine or N-acetyl-demethylphosphinothricin-L-alanyl-L-leucine, the intermediates in the biosynthesis of the tripeptides bialaphos and phosalacine (which inhibit plant and bacterial glutamine synthetases)

735668

-

?

methylcob(III)alamin

4 entries

[4Fe-4S] center

Kitasatospora phosalacinea

A0A059U454

the enzyme is a radical S-adenosyl-L-methionine enzyme that contains a [4Fe-4S] center and a methylcob(III)alamin cofactor. According to the proposed mechanism, the reduced iron-sulfur center donates an electron to S-adenosyl-L-methionine, resulting in homolytic cleavage of the carbon-sulfur bond to form a 5'-deoxyadenosyl radical that abstracts the hydrogen atom from the P-H bond of the substrate, forming a phosphinate-centered radical. This radical reacts with methylcob(III)alamin to produce the methylated product and cob(II)alamin, which is reduced by an unknown donor to cob(I)alamin. A potential route for restoring the latter back to methylcob(III)alamin is a nucleophilic attack on a second S-adenosyl-L-methionine molecule

735668

[4Fe-4S] cluster

Shewanella denitrificans

-

the enzyme binds 6.1 mol of iron per mol protein and 4.0 mol of sulfid

737755

Kitasatospora phosalacinea

2 entries

Shewanella denitrificans

-

737755

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Shewanella denitrificans OS217

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737755

-

Streptomyces hygroscopicus

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738318, 738468, 739227

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Streptomyces hygroscopicus SF-1293

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738468

-

Streptomyces viridochromogenes

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739227

-

physiological function

2 entries

64800

Shewanella denitrificans

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x * 64800, SDS-PAGE

737755

?

2 entries

-

Kitasatospora phosalacinea

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738957

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Kitasatospora phosalacinea

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738957

expression in Streptomyces lividans

Streptomyces hygroscopicus

-

738468

SD_1168 from Shewanella denitrificans is cloned into pET-30a(+)for protein overexpression. SD_1168 is found exclusively in inclusion bodies when overexpressed in either Escherichia coli or Streptomyces lividans. All efforts to express soluble recombinant protein are unsuccessful. Therefore SD_1168 is refolded using an established refolding procedure

Shewanella denitrificans

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737755

ExplorEnz (official portal for IUBMB Enzyme Nomenclature)

Expasy Enzyme Nomenclature Database

KEGG

MetaCyc

SABIO-RK

NCBI: PubMed, Protein, Nucleotide, Structure, Gene, OMIM

Enzyme Nomenclature (alternative site)

UniProt

PDB

PROSITE Database of protein families and domains

InterPro (database of protein families, domains and functional sites)