BRENDA - Reference to 1.13.11.72; Id = 745178

recombinant expression of wild-type and mutant enzymes in Escherichia coli

Streptomyces viridochromogenes

E176A

site-directed mutagenesis, the mutant enzyme shows similar activity as the wild-type enzyme. Like the wild-type enzyme, the mutant HEPD-E176A produces hydroxymethylphosphonate and formate as its only detectable products upon incubation with Fe(II), hydroxyethylphosphonate, and O2

Streptomyces viridochromogenes

additional information

-

additional information

stopped-flow and multi-turnover steady-state kinetics

Streptomyces viridochromogenes

Fe2+

required for catalysis, the mechanism involves activation of an O-H bond by the ferryl complex

Streptomyces viridochromogenes

2-hydroxyethylphosphonate + O2

Streptomyces viridochromogenes

-

hydroxymethylphosphonate + formate

-

?

2-hydroxyethylphosphonate + O2

Streptomyces viridochromogenes DSM 40736

-

hydroxymethylphosphonate + formate

-

?

Streptomyces viridochromogenes

Q5IW40

-

Streptomyces viridochromogenes DSM 40736

Q5IW40

-

recombinant wild-type and mutant enzymes from Escherichia coli

Streptomyces viridochromogenes

2-hydroxyethylphosphonate + O2 = hydroxymethylphosphonate + formate

the reaction proceeds via a transient iron(IV)-oxo (ferryl) complex, a mechanism that involves activation of an O-H bond by the ferryl complex is proposed. The isotope-sensitive C-H-cleavage step is not primarily rate-limiting for the overall catalytic cycle. The reaction does exhibit a significant 2H-kinetic isotope effect on kcat/Km for O2, which implies reversible formation of the C-H-cleaving intermediate

Streptomyces viridochromogenes

2-hydroxyethylphosphonate + O2

-

745178

Streptomyces viridochromogenes

hydroxymethylphosphonate + formate

-

?

2-hydroxyethylphosphonate + O2

the reaction proceeds via a transient iron(IV)-oxo (ferryl) complex, the mechanism involves activation of an O-H bond by the ferryl complex. Maximal accumulation of the intermediate requires both the presence of deuterium in the substrate and, importantly, the use of 2H2O as solvent

745178

Streptomyces viridochromogenes

hydroxymethylphosphonate + formate

-

?

2-hydroxyethylphosphonate + O2

-

745178

Streptomyces viridochromogenes DSM 40736

hydroxymethylphosphonate + formate

-

?

2-hydroxyethylphosphonate + O2

the reaction proceeds via a transient iron(IV)-oxo (ferryl) complex, the mechanism involves activation of an O-H bond by the ferryl complex. Maximal accumulation of the intermediate requires both the presence of deuterium in the substrate and, importantly, the use of 2H2O as solvent

745178

Streptomyces viridochromogenes DSM 40736

hydroxymethylphosphonate + formate

-

?

x * 30000, recombinant enzyme, SDS-PAGE

Streptomyces viridochromogenes

recombinant expression of wild-type and mutant enzymes in Escherichia coli

Streptomyces viridochromogenes

E176A

site-directed mutagenesis, the mutant enzyme shows similar activity as the wild-type enzyme. Like the wild-type enzyme, the mutant HEPD-E176A produces hydroxymethylphosphonate and formate as its only detectable products upon incubation with Fe(II), hydroxyethylphosphonate, and O2

Streptomyces viridochromogenes

additional information

-

additional information

stopped-flow and multi-turnover steady-state kinetics

Streptomyces viridochromogenes

Fe2+

required for catalysis, the mechanism involves activation of an O-H bond by the ferryl complex

Streptomyces viridochromogenes

2-hydroxyethylphosphonate + O2

Streptomyces viridochromogenes

-

hydroxymethylphosphonate + formate

-

?

2-hydroxyethylphosphonate + O2

Streptomyces viridochromogenes DSM 40736

-

hydroxymethylphosphonate + formate

-

?

recombinant wild-type and mutant enzymes from Escherichia coli

Streptomyces viridochromogenes

2-hydroxyethylphosphonate + O2

-

745178

Streptomyces viridochromogenes

hydroxymethylphosphonate + formate

-

?

2-hydroxyethylphosphonate + O2

the reaction proceeds via a transient iron(IV)-oxo (ferryl) complex, the mechanism involves activation of an O-H bond by the ferryl complex. Maximal accumulation of the intermediate requires both the presence of deuterium in the substrate and, importantly, the use of 2H2O as solvent

745178

Streptomyces viridochromogenes

hydroxymethylphosphonate + formate

-

?

2-hydroxyethylphosphonate + O2

-

745178

Streptomyces viridochromogenes DSM 40736

hydroxymethylphosphonate + formate

-

?

2-hydroxyethylphosphonate + O2

the reaction proceeds via a transient iron(IV)-oxo (ferryl) complex, the mechanism involves activation of an O-H bond by the ferryl complex. Maximal accumulation of the intermediate requires both the presence of deuterium in the substrate and, importantly, the use of 2H2O as solvent

745178

Streptomyces viridochromogenes DSM 40736

hydroxymethylphosphonate + formate

-

?

x * 30000, recombinant enzyme, SDS-PAGE

Streptomyces viridochromogenes

physiological function

2-hydroxyethylphosphonate dioxygenase (HEPD) cleaves the C1-C2 bond of its substrate to afford hydroxymethylphosphonate on the biosynthetic pathway to the commercial herbicide phosphinothricin

Streptomyces viridochromogenes

physiological function

2-hydroxyethylphosphonate dioxygenase (HEPD) cleaves the C1-C2 bond of its substrate to afford hydroxymethylphosphonate on the biosynthetic pathway to the commercial herbicide phosphinothricin

Streptomyces viridochromogenes

745178

Peck

O-H activation by an unexpect ...

Streptomyces viridochromogenes, Streptomyces viridochromogenes DSM 40736

J. Am. Chem. Soc.

139

2045-2052

2017

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1

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1

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1

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1

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2

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4

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1

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1

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1

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1

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1

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1

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-

1

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4

1

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1

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745390

Peck

Go it alone four-electron oxi ...

Streptomyces viridochromogenes, Streptomyces viridochromogenes DSM 40736

J. Biol. Inorg. Chem.

22

381-394

2017

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1

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1

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1

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6

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2

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745162

Peck

A common late-stage intermedi ...

Streptomyces viridochromogenes, Streptomyces viridochromogenes DSM 40736

J. Am. Chem. Soc.

137

3217-3220

2015

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1

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1

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1

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4

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1

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1

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2

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1

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720308

Du

Water-dependent reaction pathw ...

Streptomyces viridochromogenes

J. Phys. Chem. B

116

11837-11844

2012

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1

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2

-

2

-

718907

Peck

Mechanism and substrate recogn ...

Streptomyces viridochromogenes

Biochemistry

50

6598-6605

2011

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4

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6

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1

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6

719655

Whitteck

On the stereochemistry of 2-hy ...

Streptomyces viridochromogenes

J. Am. Chem. Soc.

133

4236-4239

2011

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1

-

3

-

3

-

719648

Hirao

Ferric superoxide and ferric h ...

Streptomyces viridochromogenes

J. Am. Chem. Soc.

132

17901-17909

2010

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1

-

1

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1

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1

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1

-

719645

Whitteck

Hydroperoxylation by hydroxyet ...

Streptomyces viridochromogenes

J. Am. Chem. Soc.

131

16225-16232

2009

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1

-

1

-

6

-

1

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6

-

720539

Cicchillo

An unusual carbon-carbon bond ...

Streptomyces viridochromogenes

Nature

459

871-874

2009

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1

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1

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1

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1

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1

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1

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O-H activation by an unexpected ferryl intermediate during catalysis by 2-hydroxyethylphosphonate dioxygenase

Data extracted from this reference: