Information on EC 3.5.1.137 - N-methylcarbamate hydrolase

-

-

?

3,5-dimethylphenyl N-methylcarbamate + H2O

?

3,5-dimethylphenyl N-methylcarbamate + H2O

? + methylamine + CO2

4-nitrophenyl acetate + H2O

4-nitrophenol + acetate

9 entries

acetamide + H2O

?

aldicarb + H2O

?

7 entries

aldicarb + H2O

? + methylamine + CO2

an N-methyl carbamate ester + H2O

an alcohol + methylamine + CO2

-

-

?

carbaryl + H2O

1-naphthol + methylamine + CO2

21 entries

carbofuran + H2O

2,3-dihydro-2,2-dimethyl-7-benzofuranol + methylamine + CO2

-

i.e. 2,3-dihydro-2,2-dimethyl-7-benzofuranyl N-methylcarbamate

-

?

carbofuran + H2O

7-phenol + methylamine + CO2

carbofuran + H2O

?

carbofuran + H2O

? + methylamine + CO2

carbofuran + H2O

carbofuran phenol + methylamine + CO2

dimethyl umbelliferyl phosphate + H2O

umbelliferone + dimethyl phosphate

-

-

?

fenobucarb + H2O

?

4 entries

fenobucarb + H2O

? + methylamine + CO2

formamide + H2O

?

iprovalicarb + H2O

? + methylamine + CO2

-

-

?

isoprocarb + H2O

?

9 entries

isoprocarb + H2O

? + methylamine + CO2

methomyl + H2O

methyl (1E)-N-hydroxyethanimidothioate + methylamine + CO2

methylparathion + H2O

?

i.e. O,O-dimethyl-O-4-nitrophenylthiophosphate

-

?

metolcarb + H2O

?

metolcarb + H2O

? + methylamine + CO2

oxamyl + H2O

?

oxamyl + H2O

? + methylamine + CO2

oxamyl oxime + H2O

? + methylamine + CO2

-

-

?

pirimicarb + H2O

? + methylamine + CO2

-

-

?

propoxur + H2O

?

9 entries

propoxur + H2O

? + methylamine + CO2

prosulfocarb + H2O

? + methylamine + CO2

-

-

?

xylylcarb + H2O

?

xylylcarb + H2O

? + methylamine + CO2

additional information

?

-

16 entries

4-nitrophenyl acetate + H2O

4-nitrophenol + acetate

-

-

?

aldicarb + H2O

?

-

34.74% activity compared to carbaryl

-

?

an N-methyl carbamate ester + H2O

an alcohol + methylamine + CO2

-

-

?

carbaryl + H2O

1-naphthol + methylamine + CO2

carbofuran + H2O

7-phenol + methylamine + CO2

-

51.24% activity compared to carbaryl

-

?

fenobucarb + H2O

?

-

69.84% activity compared to carbaryl

-

?

isoprocarb + H2O

?

-

i.e. 2-isopropylphenyl N-methylcarbamate, 2% activity compared to carbaryl

-

?

oxamyl + H2O

?

-

31.25% activity compared to carbaryl

-

?

propoxur + H2O

?

-

66.01% activity compared to carbaryl

-

?

additional information

?

-

-

the enzyme does not hydrolyze N-acetyl-4-aminophenol or methomyl

-

additional information

-

the enzyme does not require any cofactor

-

Ca2+

122.13% activity at 1 mM

Co2+

Cu2+

Sphingobium sp. CFD-1

-

activates

Fe2+

117.99% activity at 1 mM

Mn2+

Ni2+

Zn2+

additional information

2-chloromercuribenzoate

-

21% residual activity at 1 mM

-

2-mercaptoethanol

Pseudomonas sp. 50432

-

the enzyme shows 10% activity in the presence of 0.1% (v/v) 2-mercaptoethanol. Higher concentrations completely inactivate the enzyme. However, the activity of the enzyme is recovered when 2-mercaptoethanol was removed by dialysis

acetone

-

20% residual activity in 25% (v/v) acetone

acetonitrile

-

20% residual activity in 14% (v/v) acetonitrile

Ag+

-

complete inhibition at 1 mM

Cu2+

4 entries

diisopropylfluorophosphate

dithiothreitol

Pseudomonas sp. 50432

-

the enzyme shows 10% activity in the presence of 0.1% (v/v) dithiothreitol. Higher concentrations completely inactivate the enzyme

EDTA

Fe2+

-

complete inhibition at 1 mM

Fe3+

Hg2+

iodoacetic acid

-

13% residual activity at 1 mM

methanol

-

20% residual activity in 25% (v/v) methanol

Mg2+

-

93% residual activity at 1 mM

Mn2+

-

96% residual activity at 1 mM

paraoxon

-

less than 1% residual activity at 1 mM

Phenanthroline

-

complete inhibition at 10 mM

phenylmethylsulfonyl fluoride

Zn2+

additional information

-

dithiothreitol

-

110% activity at 1 mM

SDS

Pseudomonas sp. CRL-OK

-

hydrolase activity is slightly enhanced by low concentrations of SDS and retains greater than 70% of its activity in assay mixtures containing up to 0.5% (w/v) SDS

additional information

Pseudomonas sp. CRL-OK

-

hydrolase activity is not affected by the presence or absence of sulfhydryl reagents such as 2-mercaptoethanol (more than 90% activity in assays containing up to 0.2 mM 2-mercaptoethanol). The enzyme activity is unaffected in assays containing up to 10 mM EDTA

-

0.053 - 0.223

1-naphthyl acetate

2.7

2-nitrophenyl dimethylcarbamate

-

at pH 8.5 and 37Â°C

-

0.093

3,5-dimethylphenyl N-methylcarbamate

-

at pH 7.0 and 30Â°C

-

0.036

4-nitrophenyl acetate

-

at pH 7.0 and 30Â°C

0.542 - 2.8

Aldicarb

0.009 - 0.213

Carbaryl

18 entries

0.00053 - 0.88

carbofuran

12 entries

0.167

dimethyl umbelliferyl phosphate

-

at pH 6.8 and 37°C

-

0.587

fenobucarb

-

at pH 7.0 and 30Â°C

0.551

isoprocarb

-

at pH 7.0 and 30Â°C

0.0875

methomyl

-

at pH 7.5 and 30Â°C

0.041

metolcarb

-

at pH 7.0 and 30Â°C

-

0.1472 - 1.5803

oxamyl

0.387

propoxur

-

at pH 7.0 and 30Â°C

0.039 - 0.172

xylylcarb

-

52.7 - 906

1-naphthyl acetate

1581

3,5-dimethylphenyl N-methylcarbamate

-

at pH 7.0 and 30Â°C

-

1070

4-nitrophenyl acetate

-

at pH 7.0 and 30Â°C

548

Aldicarb

-

at pH 7.0 and 30Â°C

0.33 - 1724

Carbaryl

12 entries

0.6 - 1282

carbofuran

8 entries

0.00000051

dimethyl umbelliferyl phosphate

-

at pH 6.8 and 37°C

-

613

fenobucarb

-

at pH 7.0 and 30Â°C

670

isoprocarb

-

at pH 7.0 and 30Â°C

345.2

methomyl

-

at pH 7.5 and 30Â°C

1369

metolcarb

-

at pH 7.0 and 30Â°C

-

0.2 - 5.8

oxamyl

717

propoxur

-

at pH 7.0 and 30Â°C

1633

xylylcarb

-

at pH 7.0 and 30Â°C

-

240 - 17100

1-naphthyl acetate

17000

3,5-dimethylphenyl N-methylcarbamate

-

at pH 7.0 and 30Â°C

-

29700

4-nitrophenyl acetate

-

at pH 7.0 and 30Â°C

530

Aldicarb

-

at pH 7.0 and 30Â°C

8.7 - 144000

Carbaryl

11 entries

0.81 - 19300

carbofuran

3.1

dimethyl umbelliferyl phosphate

-

at pH 6.8 and 37°C

-

1040

fenobucarb

-

at pH 7.0 and 30Â°C

1210

isoprocarb

-

at pH 7.0 and 30Â°C

3900

methomyl

-

at pH 7.5 and 30Â°C

33400

metolcarb

-

at pH 7.0 and 30Â°C

-

0.13 - 28

oxamyl

1850

propoxur

-

at pH 7.0 and 30Â°C

41800

xylylcarb

-

at pH 7.0 and 30Â°C

-

0.013

Pseudomonas sp. CRL-OK

-

crude soluble extract, at pH 7.0, temperature not specified in the publication

0.025

-

crude extract, with 4-nitrophenyl acetate as substrate, at pH 8.3 and 25°C

0.043

-

crude extract, at pH 7.0 and 30Â°C

0.05

crude extract, at pH 7.0 and 30°C

0.14

-

crude enzyme from supernatant, at pH 8.5 and 37Â°C

0.182

-

after 7.3fold purification, with 4-nitrophenyl acetate as substrate, at pH 8.3 and 25°C

0.388

-

after 9.1fold purification, at pH 7.0 and 30Â°C

0.5

Pseudomonas sp. 50432

-

crude cell extract, at pH 7.0 and 25°C

0.68

-

crude extract, at pH 7.0 and 30°C

1016

Pseudomonas sp. 50432

-

after 2032fold purification, at pH 7.0 and 25°C

1201.8

-

after 1767fold purification, at pH 7.0 and 30°C

14.1

Pseudomonas sp. CRL-OK

-

after 1085fold purification, at pH 7.0, temperature not specified in the publication

2.3

Blastobacter sp. M501

-

crude extract, at pH 7.0 and 30°C

3.7

-

cell extract, at pH 7.0 and 30Â°C

30.7

after 614fold purification, at pH 7.0 and 30°C

7.85

-

after 58.1fold purification, at pH 8.5 and 37Â°C

704.3

Blastobacter sp. M501

-

after 306.2fold purification, at pH 7.0 and 30°C

968

-

after 262fold purification, at pH 7.0 and 30Â°C

7

7.5

-

8

-

8 - 8.5

Pseudomonas sp. CRL-OK

-

8.5

9

9 - 10.5

-

5.5 - 9

-

more than 75% activity between pH 5.5 and 9.0

6 - 10

-

the enzyme retains over 80% of its maximum activity between pH 6.0 and 10.0

6 - 11

6 - 8

the enzyme retains more than 80% activity after incubation at pH 6.0-8.0

6.5 - 9

-

the enzyme exhibits high activity levels at a pH range of 6.5 to 9.0

30

-

37

Pseudomonas sp. 50432

-

40

45

4 entries

45 - 53

-

50

60

Pseudomonas sp. CRL-OK

-

20 - 40

-

more than 50% activity between 20 and 40°C

30 - 50

more than 60% activity between 30 and 50°C

30 - 55

-

more than 50% activity between 30 and 55 C. The enzyme has 56% of its full activity at 55 C and is completely inactivated at 65 C

4.6

-

isoelectric focusing

4.8

-

isoelectric focusing

6.9

Pseudomonas sp. 50432

-

isoelectric focusing

-

763166, 763167, 763194, 763289

-

-

-

-

-

-

-

-

-

-

-

Aspergillus niger PY168

-

-

Blastobacter sp. M501

-

-

isolate 50581

-

Pseudomonas sp. 50432

-

-

-

UniProt

-

-

Pseudomonas sp. CRL-OK

-

-

Pseudomonas sp. OXA20

-

-

-

UniProt

-

762631, 763336, 763709

UniProt

Rhizobium sp. X9

-

763176

-

Sphingobium sp. CFD-1

-

762640, 763336

-

-

UniProt

cytosol

membrane

-

100000

-

gel filtration

105000

-

gel filtration

150000

-

density gradient centrifugation

160000

gel filtration

166000

Blastobacter sp. M501

-

gel filtration

170000

-

gel filtration

187000

Pseudomonas sp. CRL-OK

-

sucrose density gradient centrifugation

48000

-

gel filtration

65000

70000

88000

Pseudomonas sp. 50432

-

gel filtration

?

homodimer

monomer

D437A

-

inactive

H341A

-

inactive

H343A

-

inactive

H346A

-

inactive

H416A

-

inactive

D191A

-

the activity of the mutant enzyme is completely abolished

E207A

-

the activity of the mutant enzyme is completely abolished

G162A

-

the activity of the mutant enzyme is completely abolished

H193A

-

the mutant retains 10% of hydrolysis activity against methomyl as compared to the wild type enzyme

N164A

-

the activity of the mutant enzyme is completely abolished

D191A

-

the activity of the mutant enzyme is completely abolished

-

E207A

-

the activity of the mutant enzyme is completely abolished

-

G162A

-

the activity of the mutant enzyme is completely abolished

-

H193A

-

the mutant retains 10% of hydrolysis activity against methomyl as compared to the wild type enzyme

-

N164A

-

the activity of the mutant enzyme is completely abolished

-

F152L

A0A158SQ52

the mutation determines the catalytic preference for bicyclic carbamate substrates (carbofuran, carbaryl)

L152F

-

the mutation renders the enzyme completely inactive against carbofuran

762975

T494A

A0A158SQ52

the mutation determines the preference for monocyclic carbamate substrates (isoprocarb, propoxur)

T570I

A0A158SQ52

the mutation determines the preference for linear carbamate substrates (oxamyl, aldicarb)

F152L

Pseudomonas sp. OXA20

-

the mutation determines the catalytic preference for bicyclic carbamate substrates (carbofuran, carbaryl)

T494A

Pseudomonas sp. OXA20

-

the mutation determines the preference for monocyclic carbamate substrates (isoprocarb, propoxur)

T570I

Pseudomonas sp. OXA20

-

the mutation determines the preference for linear carbamate substrates (oxamyl, aldicarb)

H451A

the mutant shows wild type activity

H464A

the mutant shows wild type activity

H467A

the mutant shows 32.68% activity compared to the wild type enzyme

H467A/H477A

the mutant shows 7.28% activity compared to the wild type enzyme

H477A

the mutant shows 78.28% activity compared to the wild type enzyme

H504A

the mutant shows 1.34% activity compared to the wild type enzyme

F152L

the mutation determines the catalytic preference for bicyclic carbamate substrates (carbofuran, carbaryl)

T494A

the mutation determines the preference for monocyclic carbamate substrates (isoprocarb, propoxur)

T570I

the mutation determines the preference for linear carbamate substrates (oxamyl, aldicarb)

F152L

Sphingobium sp. CFD-1

A0A6B9IL91

the mutation determines the catalytic preference for bicyclic carbamate substrates (carbofuran, carbaryl)

H351A

Sphingobium sp. CFD-1

-

the mutant loses hydrolysis activity against carbofuran

H353A

Sphingobium sp. CFD-1

-

the mutant loses hydrolysis activity against carbofuran

H453A

Sphingobium sp. CFD-1

-

the mutant loses hydrolysis activity against carbofuran

H495A

Sphingobium sp. CFD-1

-

the mutant loses hydrolysis activity against carbofuran

T494A

Sphingobium sp. CFD-1

A0A6B9IL91

the mutation determines the preference for monocyclic carbamate substrates (isoprocarb, propoxur)

T570I

Sphingobium sp. CFD-1

A0A6B9IL91

the mutation determines the preference for linear carbamate substrates (oxamyl, aldicarb)

F152L

-

the mutation determines the catalytic preference for bicyclic carbamate substrates (carbofuran, carbaryl)

T494A

-

the mutation determines the preference for monocyclic carbamate substrates (isoprocarb, propoxur)

T570I

-

the mutation determines the preference for linear carbamate substrates (oxamyl, aldicarb)

10 - 11

6 - 8

7 - 8.5

-

the enzyme retains 75% of its activity after storage at pH 7.0 to 8.5 for 1 h at 30Â°C

762651

30 - 50

30 - 55

-

incubation at temperatures in excess of 45°C reduce the carbaryl hydrolase activity of the enzyme, with half maximal activity at approximately 55°C. However, incubation at between 30°C and 45°C cause an overall increase in activity

4 - 40

-

the activity of the enzyme remains above 70% after incubation at 4-40°C for 2 h. When the enzyme is incubated above 60°Cfor 2 h, it loses about 95% of its activity

4 - 50

the enzyme retains more than 80% activity after 6 h incubation at temperatures ranging from 4 to 30ºC. However, nearly 80% of the enzyme activity is lost when the temperature is higher than 50ºC for 6 h

60

Pseudomonas sp. 50432

-

when heated at 60°C for 5 min, the catalytic activity of the enzyme is completely lost

60 - 70

-

40% of the initial activity is retained after 60 min at 60Â°C. The enzyme is completely inactivated by heating for 60 min at 70Â°C

65

-

the enzyme loses its catalytic activity when heated to 65°C for 5 min

70 - 80

-

the enzyme retains more than 80% of its activity at 55Â°C for 1 h. At 70Â°C, the residual enzyme activity falls below 10% in 1 h

enzyme activity is not diminished after repeated cycles of freezing and thawing

Pseudomonas sp. CRL-OK

-

the enzyme is stable in Triton X-100 and other detergents such as CHAPS

Pseudomonas sp. 50432

-

-20 or -70°C, purified enzyme in 10 mM K2HPO4 (pH 7.5) in the presence of 20% (v/v) glycerol, 1 month, no loss of activity

Pseudomonas sp. 50432

-

-70°C, purified enzyme in the presence of 20% (v/v) glycerol, 1 month, no loss of activity

-

-80°C, crude extract and purified enzyme, more than 1 month, no loss of activity

Pseudomonas sp. CRL-OK

-

0°C, purified enzyme, 48 h, no loss of activity

-

25°C, purified enzyme, 10 days, 10% loss of activity

-

25°C, purified enzyme, 12 h, 15% loss of activity

-

25°C, purified enzyme, 25 days, 47% loss of activity

-

25°C, purified enzyme, 7 days, no loss of activity

-

4°C, after partial purification by DEAE column chromatography, more than 1 month, no loss of activity

Pseudomonas sp. CRL-OK

-

4°C, crude extract, 24 h, 30% loss of activity

Pseudomonas sp. CRL-OK

-

4°C, purified enzyme in 10 mM K2HPO4 (pH 7.5), 1 month, 25% loss of activity

Pseudomonas sp. 50432

-

4°C, purified enzyme in 10 mM K2HPO4 (pH 7.5), 1 week, no loss of activity

Pseudomonas sp. 50432

-

ammonium sulfate precipitation, Q Sepharose column chromatography, SP Sepharose column chromatography, and Sephacryl S-200 gel filtration

DEAE column chromatography, TSK phenyl column chromatography and 300SW gel filtration

Pseudomonas sp. CRL-OK

-

DEAE-cellulose column chromatography, hydroxylapatite column chromatography, and Sephacryl S-300 gel filtration

-

DEAE-cellulose column chromatography, Sephadex G-200 gel filtration, and Macro-prep tert-butyl column chromatography

Pseudomonas sp. 50432

-

His-bind resin column chromatography

His-Spin Protein Miniprep kit

-

762975

Ni-NTA agarose resin column chromatography

Ni-NTA agarose resin column chromatography and Superdex 200 gel filtration

Sphingobium sp. CFD-1

-

Ni-NTA column chromatography

-

Ni-NTA resin column chromatography and Superdex-200 gel filtration

-

phenyl Sepharose column chromatography and Superdex 200 gel filtration

-

protamine sulfate precipitation, ammonium sulfate precipitation, phenyl Sepharose column chromatography, and Biogel A0.5m gel filtration

-

protamine sulfate precipitation, ammonium sulfate precipitation, phenyl Sepharose column chromatography, DEAE-Sepharose column chromatography, and Sephacryl 200 gel filtration

-

protamine sulfate treatment, ammonium sulfate precipitation, phenyl Sepharose column chromatography, DEAE-Sepharose column chromatography, and Superdex 200 gel filtration

-

protamine sulfate treatment, ammonium sulfate precipitation, Phenyl-Sepharose CL-4B column chromatography, DEAE-Toyopearl 650 column chromatography, Toyopearl HW-55 gel filtration, and hydroxylapatite column chromatography

Blastobacter sp. M501

-

Q-Sepharose column chromatography, Sepharose CL-4B column chromatography, hydroxyapatite column chromatography and Sephacryl S-200HR gel filtration

Pseudomonas sp. C7

-

762644

Q-Sepharose column chromatography, Sepharose CL-4B column chromatography, hydroxyapatite column chromatography and Sephacryl S-200HR gel filtration, and Ni-NTA column chromatography

Pseudomonas sp. C5pp

A0A140DCH5

762644

TSK-DEAE column chromatography, TSK-phenyl column chromatography, and Bio-Gel A0.5m gel filtration

-

enzyme variant rCHDELTA(Tmd)a is expressed in Escherichia coli BL21(DE3) cells

Pseudomonas sp. C5pp

A0A140DCH5

762644

expressed in Escherichia coli BL21(DE3) cells

10 entries

expressed in Escherichia coli BL21(lambdaDE3) cells

-

expressed in Escherichia coli JM109 cells

expressed in Pseudomonas putida

-

763194

expressed on the cell surface of Stenotrophomonas sp. YC-1

763709

carbofuran stimulates the abundance and expression of the enzyme gene

oxamyl and carbofuran stimulate the abundance and expression of the enzyme gene