Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Schofield, D.A.; Dinovo, A.A.
    Generation of a mutagenized organophosphorus hydrolase for the biodegradation of the organophosphate pesticides malathion and demeton-S (2010), J. Appl. Microbiol., 109, 548-557.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.8.1 expressed in Escherichia coli BL21(DE3)pLysS cells Flavobacterium sp.

Protein Variants

EC Number Protein Variants Comment Organism
3.1.8.1 A80V/F132C/K185R/D208G/H257W/I274N/R319S the mutant shows increased specific activity with demethon-S methyl and malathion compared to the wild type enzyme Flavobacterium sp.
3.1.8.1 A80V/F132D/K185R/D208G/H257W/I274N/R319S the mutant shows increased specific activity with demethon-S methyl and malathion compared to the wild type enzyme Flavobacterium sp.
3.1.8.1 A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S the mutant shows increased specific activity with VX, demethon-S methyl and malathion compared to the wild type enzyme Flavobacterium sp.
3.1.8.1 A80V/I106V/F132D/K185R/D208G/H257W/I274N/S308L/R319S the specific activity of the mutant is improved 26, 35 and 42fold against VX, malathion and demeton-S methyl, respectively, compared to the wild type Flavobacterium sp.
3.1.8.1 A80V/K185R/D208G/H257W/I274N/R319S the mutant shows increased specific activity with demethon-S methyl and malathion compared to the wild type enzyme Flavobacterium sp.
3.1.8.1 A80V/K185R/D208G/I274N/R319S the mutant shows increased specific activity with demethon-S methyl and malathion compared to the wild type enzyme Flavobacterium sp.
3.1.8.1 G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/F306V/R319S the mutant displays a 77fold improvement in the specific activity against malathion, and exhibits only about 10% of the specific activity against VX compared to the wild type enzyme Flavobacterium sp.
3.1.8.1 G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S the mutant shows increased specific activity with demethon-S methyl and malathion compared to the wild type enzyme Flavobacterium sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.8.1 0.29
-
 malathion mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/F306V/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.41
-
 malathion wild type enzyme, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.67
-
 malathion mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/S308L/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.82
-
 malathion mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 1.16
-
 malathion mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 1.2
-
 demeton-S methyl mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/S308L/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 3.64
-
 demeton-S methyl mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 4.18
-
 demeton-S methyl mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/F306V/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 8.09
-
 demeton-S methyl wild type enzyme, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 23.31
-
 demeton-S methyl mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.1.8.1 40000
-
 x * 40000, SDS-PAGE Flavobacterium sp.

Organism

EC Number Organism UniProt Comment Textmining
3.1.8.1 Flavobacterium sp.
-
-
-
3.1.8.1 Flavobacterium sp. ATCC 27551
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.8.1 nickel agarose affinity column chromatography Flavobacterium sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.1.8.1 0.000008
-
 wild type enzyme, using malathion as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.00015
-
 wild type enzyme, using demeton-S methyl as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.00019
-
 mutant enzyme A80V/F132C/K185R/D208G/H257W/I274N/R319S, using malathion as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.00022
-
 mutant enzyme A80V/K185R/D208G/H257W/I274N/R319S, using malathion as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.00045
-
 mutant enzyme A80V/F132D/K185R/D208G/H257W/I274N/R319S, using malathion as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.00048
-
 mutant enzyme A80V/K185R/D208G/I274N/R319S, using malathion as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.00075
-
 mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, using malathion as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.00151
-
 mutant enzyme A80V/K185R/D208G/I274N/R319S, using demeton-S methyl as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.00414
-
 mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/S308L/R319S, using malathion as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.00429
-
 mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/F306V/R319S, using demeton-S methyl as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.00472
-
 mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, using demeton-S methyl as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.00509
-
 mutant enzyme A80V/K185R/D208G/H257W/I274N/R319S, using demeton-S methyl as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.00617
-
 mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, using malathion as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.007
-
 mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, using VX as substrate, in 10 mM N-Tris (hydroxymethyl-3-amino propanesulfonic acid), pH 8.0 , at 25°C Flavobacterium sp.
3.1.8.1 0.01159
-
 mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/S308L/R319S, using demeton-S methyl as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.01404
-
 mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, using demeton-S methyl as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.01511
-
 mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/F306V/R319S, using malathion as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.0194
-
 mutant enzyme A80V/F132D/K185R/D208G/H257W/I274N/R319S, using demeton-S methyl as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.02677
-
 mutant enzyme A80V/F132C/K185R/D208G/H257W/I274N/R319S, using demeton-S methyl as substrate, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 0.0803
-
 mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, using VX as substrate, in 10 mM N-Tris (hydroxymethyl-3-amino propanesulfonic acid), pH 8.0 , at 25°C Flavobacterium sp.
3.1.8.1 0.097
-
 wild type enzyme, using VX as substrate, in 10 mM N-Tris (hydroxymethyl-3-amino propanesulfonic acid), pH 8.0 , at 25°C Flavobacterium sp.
3.1.8.1 2.5
-
 mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/F306V/R319S, using VX as substrate, in 10 mM N-Tris (hydroxymethyl-3-amino propanesulfonic acid), pH 8.0 , at 25°C Flavobacterium sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.8.1 acephate + H2O
-
 Flavobacterium sp. ?
-
 ?
3.1.8.1 acephate + H2O
-
 Flavobacterium sp. ATCC 27551 ?
-
 ?
3.1.8.1 demeton-S methyl + H2O
-
 Flavobacterium sp. 2-ethylsulfanyl-ethanethiol + dimethyl phosphate
-
 ?
3.1.8.1 malathion + H2O
-
 Flavobacterium sp. O,O-diethyl phosphorothioate + diethyl 2-mercaptosuccinate
-
 ?
3.1.8.1 paraoxon + H2O
-
 Flavobacterium sp. 4-nitrophenol + diethyl phosphate
-
 ?
3.1.8.1 paraoxon + H2O
-
 Flavobacterium sp. ATCC 27551 4-nitrophenol + diethyl phosphate
-
 ?
3.1.8.1 phosalone + H2O
-
 Flavobacterium sp. ?
-
 ?
3.1.8.1 sarin + H2O
-
 Flavobacterium sp. ?
-
 ?
3.1.8.1 sarin + H2O
-
 Flavobacterium sp. ATCC 27551 ?
-
 ?
3.1.8.1 soman + H2O
-
 Flavobacterium sp. ?
-
 ?
3.1.8.1 soman + H2O
-
 Flavobacterium sp. ATCC 27551 ?
-
 ?
3.1.8.1 tabun + H2O
-
 Flavobacterium sp. ?
-
 ?
3.1.8.1 tabun + H2O
-
 Flavobacterium sp. ATCC 27551 ?
-
 ?
3.1.8.1 VX + H2O i.e. O-ethyl S-[2-diisopropylaminoethyl]methylphosphonothiolate Flavobacterium sp. ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.1.8.1 ? x * 40000, SDS-PAGE Flavobacterium sp.

Synonyms

EC Number Synonyms Comment Organism
3.1.8.1 OPH
-
 Flavobacterium sp.
3.1.8.1 organophosphorus hydrolase
-
 Flavobacterium sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.8.1 2.55
-
 malathion wild type enzyme, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 13.16
-
 demeton-S methyl wild type enzyme, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 14.62
-
 demeton-S methyl mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/F306V/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 23.01
-
 malathion mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 40.63
-
 malathion mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 44.92
-
 malathion mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/F306V/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 45.95
-
 demeton-S methyl mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 48.27
-
 demeton-S methyl mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/S308L/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 69.44
-
 demeton-S methyl mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 73.29
-
 malathion mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/S308L/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1.8.1 1.63
-
 demeton-S methyl wild type enzyme, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 1.97
-
 demeton-S methyl mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 3.5
-
 demeton-S methyl mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/F306V/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 6.21
-
 malathion wild type enzyme, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 19.08
-
 demeton-S methyl mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 19.83
-
 malathion mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 40.23
-
 demeton-S methyl mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/S308L/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 49.54
-
 malathion mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 109.4
-
 malathion mutant enzyme A80V/I106V/F132D/K185R/D208G/H257W/I274N/S308L/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.
3.1.8.1 154.9
-
 malathion mutant enzyme G60V/A80V/I106V/F132D/K185R/D208G/H257W/I274N/F306V/R319S, in 50 mM HEPES buffer, pH 7.5, at pH, 25°C Flavobacterium sp.