Information on EC 1.3.1.10 - enoyl-[acyl-carrier-protein] reductase (NADPH, Si-specific)

1-cyclohexenylcarbonyl-CoA + NADPH

1-cyclohexylcarbonyl-CoA + NADP+

-

reaction in ansatrienin biosynthesis out of shikimic acid

-

?

2-decenoyl-[acyl-carrier protein] + NADPH

decanoyl-[acyl-carrier protein] + NADP+

-

syn addition of hydrogen via a 2-Re,3-Si attack on the double bond

390496

-

390496

?

2-hexenoyl-[acyl-carrier protein] + NADPH

hexanoyl-[acyl-carrier protein] + NADP+

2-trans-hexenoyl-CoA + NADPH

?

-

assay at pH 7.5, 23Â°C

703756

-

?

5-hydroxycyclohex-1-enecarbonyl-CoA + NADPH

5-hydroxycyclohexylcarbonyl-CoA + NADP+

-

reaction in ansatrienin biosynthesis out of shikimic acid

-

?

acyl-[acyl-carrier protein] + NADP+

trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH + H+

an acyl-[acyl-carrier protein] + NADP+

a trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH + H+

5 entries

beta-ketoacyl-[acyl-carrier protein] + NADH + H+

beta-hydroxyacyl-[acyl-carrier protein] + NAD+

-

-

?

crotonyl-CoA + NADPH + H+

?

crotonyl-N-acetyl-cysteamine + NADPH

butyryl-N-acetyl-cysteamine + NADP+

crotonyl-N-acetyl-cysteamine + NADPH + H+

butyryl-N-acetyl-cysteamine + NADP+

-

-

?

crotonyl-[acyl-carrier protein] + NADPH

butyryl-[acyl-carrier protein] + NADP+

7 entries

crotonyl-[acyl-carrier protein] + NADPH + H+

?

crotonyl-[acyl-carrier protein] + NADPH + H+

butyryl-[acyl-carrier protein] + NADP+

crotonyl-[Staphylococcus aureus acyl carrier protein] + NADPH + H+

butyryl-[Staphylococcus aureus acyl carrier protein] + NADP+

-

-

?

dodec-2-enoyl-CoA + NADPH + H+

dodecanoyl-CoA + NADP+

-

-

r

dodecenoyl-CoA + NADPH + H+

dodecanoyl-CoA + NADP+

-

-

?

dodecenoyl-N-acetyl-cysteamine + NADPH + H+

dodecanoyl-N-acetyl-cysteamine + NADP+

-

-

?

dodecenoyl-[Staphylococcus aureus acyl carrier protein] + NADPH + H+

dodecanoyl-[Staphylococcus aureus acyl carrier protein] + NADP+

-

-

?

hexanoyl-CoA + NADH + H+

?

-

-

?

hexanoyl-[acyl-carrier protein] + NADH + H+

?

-

-

?

octenoyl-N-acetyl-cysteamine + NADPH

octanoyl-N-acetyl-cysteamine + NADP+

S-((2E)-oct-2-enoyl)-N-acetylcysteamine + NADPH + H+

S-octanoyl-N-acetylcysteamine + NADP+

trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH

acyl-[acyl-carrier protein] + NADP+

4 entries

trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH + H+

acyl-[acyl-carrier protein] + NADP+

-

prefers acyl carrier protein substrates carrying fatty acids with long acyl chains

-

?

trans-2-hexenoyl-CoA + NADPH

hexanoyl-CoA + NADP+

-

-

?

trans-3,4-dihydroxycyclohexa-1,5-dienecarbonyl-CoA + NADPH

trans-4,5-dihydroxycyclohexa-2-enecarbonyl-CoA + NADP+

-

reaction in ansatrienin biosynthesis out of shikimic acid

-

?

additional information

?

-

32 entries

1-cyclohexenylcarbonyl-CoA + NADPH

1-cyclohexylcarbonyl-CoA + NADP+

-

reaction in ansatrienin biosynthesis out of shikimic acid

-

?

5-hydroxycyclohex-1-enecarbonyl-CoA + NADPH

5-hydroxycyclohexylcarbonyl-CoA + NADP+

-

reaction in ansatrienin biosynthesis out of shikimic acid

-

?

an acyl-[acyl-carrier protein] + NADP+

a trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH + H+

beta-ketoacyl-[acyl-carrier protein] + NADH + H+

beta-hydroxyacyl-[acyl-carrier protein] + NAD+

-

-

?

trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH

acyl-[acyl-carrier protein] + NADP+

trans-3,4-dihydroxycyclohexa-1,5-dienecarbonyl-CoA + NADPH

trans-4,5-dihydroxycyclohexa-2-enecarbonyl-CoA + NADP+

-

reaction in ansatrienin biosynthesis out of shikimic acid

-

?

additional information

?

-

22 entries

FAD

-

enzyme contains 0.8 mol FAD/enzyme, but functional role is not investigated

FMN

-

NADH

NADP+

NADPH

additional information

-

(E)-N-(1,2-dimethyl-1H-indol-3-ylmethyl)-N-methyl-3-(7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl)acrylamide

-

5-chloro-2-phenoxyphenol

-

5-ethyl-2-phenoxyphenol

-

6-methyl-2-(propane-1-sulfonyl)-4a,7a-dihydro-2H-thieno[3,2-d][1,2,3]diazaborinin-1-ol

-

diazaborine derivative 2b18, binds only in presence of NADH or NADPH, non-competitive inhibition, 0.2 mM lead to 25% inhibition, 0.52 mM to 50%, 1.56 mM to 80%. Increasing the inhibitor concentrations lead to preference for shorter acyl chain lengths

AFN-1252

-

aquastatin A

-

a natural inhibitor from the fungus Sporothrix sp. strain FN611, mixed-type inhibition against FabI with respect to the substrate trans-2-octenoyl N-acetylcysteamine and NADPH, prevents the growth of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus with minimum inhibitory concentration of 0.016-0.032 mg/ml, overview

degalactosylated aquastatin A

-

a natural inhibitor from the fungus Sporothrix sp. strain FN611

Hexachlorophene

Q9RMI3

half-maximal inhibition at 0.004 mM

iodoacetate

-

N-ethylmaleimide

-

NADPH

-

p-hydroxymercuribenzoate

-

390499, 390500

palmitoyl-CoA

-

0.0016 mM lead to 50% inhibition, 0.002 mM to 60%, 0.005 mM to 20%, 0.01 mM to nearly total inhibition

triclosan

4 entries

additional information

-

not inhibited by: triclosan

-

NH4Cl

required for stability and activity. Assay in presence of 0.1 M NH4Cl

0.025

1-cyclohexenylcarbonyl-CoA

-

vmax 0.0075 mmol/mg/min

0.03

5-hydroxycyclohex-1-enecarbonyl-CoA

-

vmax 0.0053 mmol/mg/min

0.008 - 8

Crotonyl-N-acetyl-cysteamine

0.0478

crotonyl-[acyl-carrier protein]

pH 7.5, 25Â°C

0.0115

crotonyl-[Staphylococcus aureus acyl carrier protein]

-

wild type enzyme, at 25Â°C, in 100 mM Na2HPO4 buffer (pH 7.8)

0.0241

dodec-2-enoyl-CoA

-

wild type enzyme, at 25Â°C, in 100 mM Na2HPO4 buffer (pH 7.8)

0.0233

dodecenoyl-N-acetyl-cysteamine

-

wild type enzyme, at 25Â°C, in 100 mM Na2HPO4 buffer (pH 7.8)

0.0045 - 0.0184

dodecenoyl-[Staphylococcus aureus acyl carrier protein]

0.016 - 1

NADPH

6 entries

additional information

NADPH

6 entries

1.5

Crotonyl-N-acetyl-cysteamine

-

wild type enzyme, at 25Â°C, in 100 mM Na2HPO4 buffer (pH 7.8)

0.038 - 0.51

crotonyl-[acyl-carrier protein]

11.6

crotonyl-[Staphylococcus aureus acyl carrier protein]

-

wild type enzyme, at 25Â°C, in 100 mM Na2HPO4 buffer (pH 7.8)

18

dodec-2-enoyl-CoA

-

wild type enzyme, at 25Â°C, in 100 mM Na2HPO4 buffer (pH 7.8)

30.4

dodecenoyl-N-acetyl-cysteamine

-

wild type enzyme, at 25Â°C, in 100 mM Na2HPO4 buffer (pH 7.8)

18.4 - 157.3

dodecenoyl-[Staphylococcus aureus acyl carrier protein]

130.2

NADPH

-

wild type enzyme, at 25Â°C, in 100 mM Na2HPO4 buffer (pH 7.8)

0.00006

(E)-N-(1,2-dimethyl-1H-indol-3-ylmethyl)-N-methyl-3-(7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl)acrylamide

pH 7.0, 30Â°C

0.000004 - 0.000069

AFN-1252

0.0014 - 0.00161

aquastatin A

0.109

NADPH

pH 7.5, 25Â°C

0.0032

aquastatin A

Staphylococcus aureus

-

FabI, pH 6.5, 30Â°C

0.0034

degalactosylated aquastatin A

Staphylococcus aureus

-

FabI, pH 6.5, 30Â°C

0.016

triclosan

Bacillus subtilis

P71079

-

390505

0.008

-

crude cell extract

0.066

-

0.18

0.2

-

purified recombinant mutant Y79N

0.3

0.4

-

purified by chromatography on DEAE-cellulose and Blue sepharose

0.882

-

crude Escherichia coli cell extract after overexpression

2.31

-

20

-

purified recombinant wild-type enzyme

3.667

-

purified, overexpressed enzyme

6

6.5

-

assay at

7.5

6

23

-

assay at

25

assay at

30

-

assay at

-

-

Bacillus cereus DSM 31

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SwissProt

Bacillus subtilis 168

-

-

enzyme Etr1p

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safflower, enoyl-ACP reductase II, as there is no information concerning the stereochemistry of hydrogen transfer from NADPH to substrate an appointment to EC 1.3.1.10 or EC 1.3.1.39 is impossible

285669

-

-

390496, 390498, 390499, 390500, 390501, 390503, 390504, 390506, 390507

-

-

-

-

Porphyromonas gingivalis W83

UniProt

-

UniProt

Salmonella enterica subsp. enterica serovar Typhimurium

4 entries

-

-

-

-

-

-

Escherichia coli BL21(DE3)

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malfunction

metabolism

Drosophila melanogaster

Staphylococcus aureus subsp. aureus MRSA252

13 entries

Staphylococcus aureus subsp. aureus N315

Thermus thermophilus HB8

27770

Salmonella enterica subsp. enterica serovar Typhimurium

-

EnvM, gene sequence

390506

29700

-

ChcA, gene sequence

30000

-

SDS-PAGE

33000

-

FabI, SDS-PAGE

33090

x * 33090, calculated

34000

-

FabK, gene sequence

37000

-

ChcA, SDS-PAGE, +/-2000

75000

-

ChcA, gel filtration on superose 12, +/-10000

?

dimer

tetramer

FabL in apo form and in the ternary complex with NADP+ and an indole naphthyridinone inhibitor, X-ray diffraction structure determination and analysis

FabL in apo form and in the ternary complex with NADP+ and inhibitor (E)-N-(1,2-dimethyl-1-H-indol-3-ylmethyl)-N-methyl-3-(7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-3-yl) acrylamide, 8 mg/ml protein in 20 mM Tris, pH 8.0, 100 mM NaCl, and 1 mM DTT, is mixed with an equal volume of reservoir solution containing 0.1 M sodium citrate pH 5.6, 8% w/v PEG 10,000 and 0.4 M magnesium acetate, hanging-drop methods, for the ternary BcFabL-NADP+-INH complex, NADP+ and inhibitor are added at the molar ratio of 1:1.5 and 1:5, respectively, equilibration in a stabilizing solution containing 0.1 M sodium citrate, pH 5.6, 8% w/v PEG 10,000, 0.4 M magnesium acetate with 30% v/v ethylene glycol as the cryoprotectant, X-ray diffraction structure determination and analysis at 2.2-3.0 A resolution

hanging drop vapour diffusion method with 0.04 M MgCl2, 0.05 M sodium cacodylate pH 6.0 and 11% (v/v) 2-methyl-2,4-pentanediol at 4Â°C

-

684170

purified recombinant wild-type enzyme free and in complex with NADPH, mutant enzyme Y79N, hanging drop vapour diffusion method, 22Â°C, 15-25 mg/ml free enzyme in 50 mM sodium phosphate, pH 7.0, 150 mM NaCl with or without 10 mM NADPH, in a 1:1 ratio with reservoir solution containing 1.9 M ammonium sulfate, and 0.1 M N-(2-acetamido)-2-iminodiacetic acid/NaOH, at pH 6.5 for the free enzyme and pH 7.0 for the cofactor-complexed enzyme, mutant Y79N is crystallized using 15 mg/ml protein and reservoir solution at pH 7.0, labeling by soaking in heavy-atom-solutions, X-ray diffraction structure determination and analysis at 1.7 A, 2.25 A, and 2.6 A, respectively

-

Y79N

-

site-directed mutagenesis, mutant shows native fold, but only 0.1% of the wild-type enzyme activity, no rescue of the enzyme-deficient mutant strain mrf1DELTA

G93S

-

leads to diazaborine resistance

390503, 390506, 390507

G93V

-

leads to triclosan resistance

390504

S241F

-

leads to temperature sensitive growth and abolished activity

390503, 390507

Y73N

P38071

site-directed mutagenesis, mutant shows native fold, but only residual wild-type enzyme activity, no rescue of the enzyme-deficient mutant strain mrf1DELTA

Salmonella enterica subsp. enterica serovar Typhimurium

G93S

-

leads to diazaborine resistance

390503, 390506

A95V

-

the mutant correlates with resistance to diphenyl ethers and causes a significant reduction in the affinity of the inhibitors for the enzyme

F204S

-

inactive, the mutant correlates with resistance to diphenyl ethers and causes a significant reduction in the affinity of the inhibitors for the enzyme

I193S

-

the mutant correlates with resistance to diphenyl ethers and causes a significant reduction in the affinity of the inhibitors for the enzyme

K41N

-

strongly decreased activity for NADPH and increased activity with NADH

M99T

-

the mutant is resistant toward inhibition by AFN-1252

R40Q

-

strongly decreased activity for NADPH and increased activity with NADH

R40Q/K41N

-

strongly decreased activity for NADPH and increased activity with NADH

Y147F

-

the mutant is catalytically defective but retains its sensitivity to inhibitor AFN-1252

Y147H

-

the mutant is catalytically defective but retains its sensitivity to inhibitor AFN-1252

M99T

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the mutant is resistant toward inhibition by AFN-1252

-

Y147F

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the mutant is catalytically defective but retains its sensitivity to inhibitor AFN-1252

-

Y147H

-

the mutant is catalytically defective but retains its sensitivity to inhibitor AFN-1252

-

additional information

7.6

-

unstable at

390499, 390500

55

-

5 min stable

60

-

5 min, inactivation

enzyme can be concentrated to 20 mg/ml without precipitation

dimethyl sulfoxide

-20Â°C, 10 mM phosphate buffer, pH 6.5, 5 mM 2-mercaptoethanol, inactivation in 2-3 months

-

-20Â°C, 30% glycerol, no significant loss of activity for at least 1 month

-20Â°C, rapidly inactivated

-

chromatography of protein overexpressed in Escherichia coli by using Sephadex G-100, DEAE, Mono-Q HR 5/5

-

in BisTris puffer pH 6.5 in order to retain NADPH dependent activity which is lost at pH 7.5

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Ni-NTA column chromatography and Superdex 200 gel filtration

-

Ni2+ affinity chromatography on his-tagged protein

Ni2+ chelation chromatography

P71079

390505

Ni2+-NTA resin chromatography

-

nickel-chelated Hi-Trap column chromatography, Hi-Trap Blue HP column chromatography, Q-Sepharose column chromatography, and Superdex-75 gel filtration

-

684170

no separation from NADH specific enzyme

-

390499, 390500

recombinant His6-tagged BcFabL from Escherichia coli strain BL21(DE3) by nickel affinity and adsorption chromatography and gel filtration

recombinant protein

recombinant wild-type enzyme and mutant Y73N from mutant strain BJ1991 mrf1DELTA

P38071

recombinant wild-type enzyme and mutant Y79N from Saccharomyces cerevisiae mutant strain BJ1991 mrf1DELTA

-

sonification and chromatography using DEAE-Cellulose, Sephadex G-100, Hydroxylapatite

-

which also catalyzes the reduction of enoyl-CoA substrates

-

390496

BcFabL from strain 6A5 is expressed in Escherichia coli strain BL21(DE3) as His6-tagged protein

chcA in pUC18 and pET3C

-

envM in bacteriophage P1

Salmonella enterica subsp. enterica serovar Typhimurium

-

390506

envM in plasmid

-

390506

expressed in Escherichia coli BL21(DE3) cells

-

684170

expressed in Escherichia coli BL21(DE3)pLysS cells

-

expressed in Escherichia coli JP1111 cells

-

expression in Escherichia coli

fabI in pUC118

-

fabI of strain RN4220 found by homology and cloned into pET15b for his-tagged overexpression

Q9RMI3

390504

fabK in expression plasmid

-

fabL in pET15b for his-tagged overexpression

P71079

390505

overexpression of wild-type enzyme and mutant Y73N in enzyme-deficient mutant strain BJ1991 mrf1DELTA, using the catalase 1 promotor

P38071

overexpression of wild-type enzyme and mutant Y79N in enzyme-deficient Saccharomyces cerevisiae mutant strain BJ1991 mrf1DELTA, using the catalase 1 promotor

-