BRENDA - Enzyme Database show
show all sequences of 1.11.1.23

Purification and characterization of the epoxidase catalyzing the formation of fosfomycin from Pseudomonas syringae

Munos, J.W.; Moon, S.J.; Mansoorabadi, S.O.; Chang, W.; Hong, L.; Yan, F.; Liu, A.; Liu, H.W.; Biochemistry 47, 8726-8735 (2008)

Data extracted from this reference:

Application
Application
Commentary
Organism
medicine
fosfomycin is a clinically useful antibiotic for the treatment of limb-threatening diabetic foot infections and lower urinary tract infections. It is effective against ciprofloxacin-resistant Escherichia coli, as well as methicillin-resistant and vancomycin-resistant strains of Staphylococcus aureus. The antimicrobial activity of fosfomycin is due to the inactivation of UDP-GlcNAc-3-O-enolpyruvyltransferase, which catalyzes the first committed step in the biosynthesis of peptidoglycan, the main component of the bacterial cell wall
Pseudomonas syringae
Cloned(Commentary)
Commentary
Organism
expression in Escherichia coli
Pseudomonas syringae
Inhibitors
Inhibitors
Commentary
Organism
Structure
EDTA
5 mM, abolishes activity
Pseudomonas syringae
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Iron
mononuclear non-heme iron-containing enzyme. Contains about one iron per monomer. Substrate and NO can bind to the ferrous center of the reduced HppE enzyme to form a stable complex
Pseudomonas syringae
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
21000
-
3 * 21000, SDS-PAGE
Pseudomonas syringae
21315
-
3 * 21315, calculated from sequence
Pseudomonas syringae
73000
-
gel filtration
Pseudomonas syringae
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(S)-2-hydroxypropylphosphonic acid + 2 NADH + O2
Pseudomonas syringae
-
cis-(1R,2S)-epoxypropylphosphonic acid + 2 H2O + 2 NAD+
i.e. fosfomycin
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Pseudomonas syringae
-
-
-
Posttranslational Modification
Posttranslational Modification
Commentary
Organism
side-chain modification
posttranslationally modified by the hydroxylation of Tyr95
Pseudomonas syringae
Purification (Commentary)
Commentary
Organism
-
Pseudomonas syringae
Reaction
Reaction
Commentary
Organism
(S)-2-hydroxypropylphosphonate + H2O2 = (1R,2S)-1,2-epoxypropylphosphonate + 2 H2O
electron transfer is presumed to be the predominant rate-limiting step. Proposed mechanism: the reaction likely begins with hydrogen atom abstraction from the C-1 position by an activated oxygen species. The C-1 centered radical intermediate can then cyclize to form fosfomycin and the reduced iron center
Pseudomonas syringae
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(S)-2-hydroxypropylphosphonic acid + 2 NADH + O2
-
702244
Pseudomonas syringae
cis-(1R,2S)-epoxypropylphosphonic acid + 2 H2O + 2 NAD+
i.e. fosfomycin
-
-
?
(S)-2-hydroxypropylphosphonic acid + 2 NADH + O2
if either iron, FMN, or NADH is omitted from the reaction mixture, no product formation is detected
702244
Pseudomonas syringae
cis-(1R,2S)-epoxypropylphosphonic acid + 2 H2O + 2 NAD+
i.e. fosfomycin
-
-
?
additional information
the enzyme also forms 2-oxopropylphosphonic acid from (R)-2-hydroxypropylphosphonic acid, the rate of this reaction is 1.9times faster than the reaction with (S)-2-hydroxypropylphosphonic acid
702244
Pseudomonas syringae
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
trimer
3 * 21000, SDS-PAGE; 3 * 21315, calculated from sequence
Pseudomonas syringae
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.0028
-
(S)-2-hydroxypropylphosphonic acid
pH 7.5
Pseudomonas syringae
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Pseudomonas syringae
Cofactor
Cofactor
Commentary
Organism
Structure
FMN
if either iron, FMN, or NADH is omitted from the reaction mixture, no product formation is detected. Due to the fact that the reduction of iron requires single electron transfer and NADH is an obligate two-electron donor, FMN is required to mediate the transfer of reducing equivalents from NADH to the active site iron
Pseudomonas syringae
NADH
NADH is essential for catalysis
Pseudomonas syringae
Application (protein specific)
Application
Commentary
Organism
medicine
fosfomycin is a clinically useful antibiotic for the treatment of limb-threatening diabetic foot infections and lower urinary tract infections. It is effective against ciprofloxacin-resistant Escherichia coli, as well as methicillin-resistant and vancomycin-resistant strains of Staphylococcus aureus. The antimicrobial activity of fosfomycin is due to the inactivation of UDP-GlcNAc-3-O-enolpyruvyltransferase, which catalyzes the first committed step in the biosynthesis of peptidoglycan, the main component of the bacterial cell wall
Pseudomonas syringae
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli
Pseudomonas syringae
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FMN
if either iron, FMN, or NADH is omitted from the reaction mixture, no product formation is detected. Due to the fact that the reduction of iron requires single electron transfer and NADH is an obligate two-electron donor, FMN is required to mediate the transfer of reducing equivalents from NADH to the active site iron
Pseudomonas syringae
NADH
NADH is essential for catalysis
Pseudomonas syringae
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
EDTA
5 mM, abolishes activity
Pseudomonas syringae
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Iron
mononuclear non-heme iron-containing enzyme. Contains about one iron per monomer. Substrate and NO can bind to the ferrous center of the reduced HppE enzyme to form a stable complex
Pseudomonas syringae
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
21000
-
3 * 21000, SDS-PAGE
Pseudomonas syringae
21315
-
3 * 21315, calculated from sequence
Pseudomonas syringae
73000
-
gel filtration
Pseudomonas syringae
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
(S)-2-hydroxypropylphosphonic acid + 2 NADH + O2
Pseudomonas syringae
-
cis-(1R,2S)-epoxypropylphosphonic acid + 2 H2O + 2 NAD+
i.e. fosfomycin
-
?
Posttranslational Modification (protein specific)
Posttranslational Modification
Commentary
Organism
side-chain modification
posttranslationally modified by the hydroxylation of Tyr95
Pseudomonas syringae
Purification (Commentary) (protein specific)
Commentary
Organism
-
Pseudomonas syringae
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
(S)-2-hydroxypropylphosphonic acid + 2 NADH + O2
-
702244
Pseudomonas syringae
cis-(1R,2S)-epoxypropylphosphonic acid + 2 H2O + 2 NAD+
i.e. fosfomycin
-
-
?
(S)-2-hydroxypropylphosphonic acid + 2 NADH + O2
if either iron, FMN, or NADH is omitted from the reaction mixture, no product formation is detected
702244
Pseudomonas syringae
cis-(1R,2S)-epoxypropylphosphonic acid + 2 H2O + 2 NAD+
i.e. fosfomycin
-
-
?
additional information
the enzyme also forms 2-oxopropylphosphonic acid from (R)-2-hydroxypropylphosphonic acid, the rate of this reaction is 1.9times faster than the reaction with (S)-2-hydroxypropylphosphonic acid
702244
Pseudomonas syringae
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
trimer
3 * 21000, SDS-PAGE; 3 * 21315, calculated from sequence
Pseudomonas syringae
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.0028
-
(S)-2-hydroxypropylphosphonic acid
pH 7.5
Pseudomonas syringae
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Pseudomonas syringae
Other publictions for EC 1.11.1.23
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
729898
Huang
Mechanistic consequences of ch ...
Streptomyces wedmorensis
J. Am. Chem. Soc.
136
2944-2947
2014
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727328
Milaczewska
On the catalytic mechanism of ...
Streptomyces wedmorensis
Chemistry
19
771-781
2013
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727696
Chang
Reaction of HppE with substrat ...
Streptomyces wedmorensis
J. Am. Chem. Soc.
135
8153-8156
2013
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730966
Wang
Evidence that the fosfomycin-p ...
Yersinia pseudotuberculosis
Science
342
991-995
2013
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727675
Huang
Evidence for radical-mediated ...
Streptomyces wedmorensis
J. Am. Chem. Soc.
134
16171-16174
2012
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702244
Munos
Purification and characterizat ...
Pseudomonas syringae
Biochemistry
47
8726-8735
2008
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702547
Schweifer
On the conversion of structura ...
Streptomyces fradiae
Bioorg. Med. Chem. Lett.
18
3056-3059
2008
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702211
Yan
Determination of the substrate ...
Streptomyces wedmorensis
Biochemistry
46
12628-12638
2007
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702208
Yan
Biosynthesis of fosfomycin, re ...
Streptomyces wedmorensis
Biochemistry
45
11473-11481
2006
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701481
Cameron
Initiating a crystallographic ...
Streptomyces wedmorensis
Acta Crystallogr. Sect. F
61
534-536
2005
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701900
Yan
Site-directed mutagenesis and ...
Streptomyces wedmorensis
Arch. Biochem. Biophys.
442
82-91
2005
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705876
Higgins
Structural insight into antibi ...
Streptomyces wedmorensis
Nature
437
838-844
2005
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706480
McLuskey
Structure and reactivity of hy ...
Streptomyces wedmorensis
Proc. Natl. Acad. Sci. USA
102
14221-14226
2005
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704157
Liu
Oxygenase activity in the self ...
Streptomyces wedmorensis
J. Am. Chem. Soc.
126
10306-10312
2004
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702205
Liu
Biochemical and spectroscopic ...
Streptomyces wedmorensis
Biochemistry
42
11577-11586
2003
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701694
Zhao
Mechanistic studies of HPP epo ...
Streptomyces wedmorensis
Angew. Chem. Int. Ed. Engl.
41
4529-4532
2002
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704155
Liu
Protein purification and funct ...
Streptomyces wedmorensis
J. Am. Chem. Soc.
123
4619-4620
2001
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701777
Watanabe
A phosphonate-induced gene whi ...
Penicillium decumbens
Appl. Environ. Microbiol.
65
1036-1044
1999
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702728
Kuzuyama
Cloning and expression in Esch ...
Pseudomonas syringae, Pseudomonas syringae PB-5123
Biosci. Biotechnol. Biochem.
63
2222-2224
1999
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5
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705742
Hidaka
Cloning and nucleotide sequenc ...
Streptomyces wedmorensis 144-91, Streptomyces wedmorensis
Mol. Gen. Genet.
249
274-280
1995
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