Information on EC 5.3.3.2 - isopentenyl-diphosphate DELTA-isomerase

-

about 2% of the activity with isopenteyl diphosphate

-

(Z)-3-(fluoromethyl)-2-buten-1-yl diphosphate

?

-

about 2% of the activity with isopenteyl diphosphate

-

3-(fluoromethyl)-3-buten-1-yl diphosphate

?

-

competition between isomerization and alkylation of the flavin cofactor

-

?

3-cyclopropyl-3-buten-1-yl diphosphate

(2Z)-3-cyclopropylbut-2-en-1-yl diphosphate

-

746921

-

r

3-Ethylbut-3-enyl diphosphate

trans-3-Methylpent-3-enyl diphosphate

-

ir

-

-

3-methylene-4-penten-1-yl diphosphate

?

-

competition between isomerization and alkylation of the flavin cofactor

-

?

Isopentenyl diphosphate

?

isopentenyl diphosphate

dimethylallyl diphosphate

81 entries

isopentenyl diphosphate

dimethylallyl phosphate

-

-

?

pyruvate

lactate

-

-

?

trans-3-Methylpent-3-enyl diphosphate

trans-3-Methylpent-2-enyl diphosphate

-

r

-

-

additional information

?

-

13 entries

Isopentenyl diphosphate

?

isopentenyl diphosphate

dimethylallyl diphosphate

25 entries

additional information

?

-

6 entries

FMN

16 entries

FMNH2

-

747099, 747530

NAD(P)H

NADH

-

or NADPH required under aerobic, not under anaerobic conditions

NADPH

8 entries

additional information

-

IDI showed almost the same activity in the presence of NADH or dithionite. Data indicate that the reduced form of FMN is sufficient

-

Ca2+

Cd2+

-

reconstitution of metal-free enzyme with Mg2+, Mn2+, Zn2+, Co2+, Ni2+ or Cd2+ generates active enzyme

Co2+

Cu2+

-

10mM, relative activity under 0.005%

Mg2+

Mn2+

Ni2+

Zinc

-

essential cofactor. Type I enzyme contains an atom of tinc. The metal is located in an unusual six-coordinate pocket and may facilitate protonation of the unactivated carbon-carbon double bond in isopentenyl diphosphate

651597

Zn2+

additional information

(+/-)-cis/trans-Chrysanthemyl diphosphate

-

(+/-)-cis/trans-Chrysanthemyl monophosphate

-

(2Z)-3-methylhept-2-en-1-yl trihydrogen diphosphate

(2Z)-3-methylhex-2-en-1-yl trihydrogen diphosphate

(2Z)-3-methyloct-2-en-1-yl trihydrogen diphosphate

(E/Z)-3-cyclopropyl-2-buten-1-yl diphosphate

-

(N,N-dimethylamino)-1-ethyl diphosphate

-

transition state analogue

(Z)-3-difluoromethyl-2-buten-1-yl diphosphate

-

reversible

(Z)-3-fluoromethyl-2-buten-1-yl diphosphate

-

reversible

(Z)-4-Fluoro-3-methyl-2-buten-1-yl diphosphate

-

inactivates through an active-site-directed covalent modification

2943

2,2-Diphenyl-1-(beta-dimethylaminoethoxy)pentane hydrochloride

-

2,3-butadien-1-yl diphosphate

2,3-cis,6,7-trans-farnesyl diphosphate

-

2-(dimethylamino)ethyl diphosphate

-

weak competitive inhibitor

746921

-

2-(dimethylamino)ethyl phosphate

2-[ethyl(methyl)amino]ethyl trihydrogen diphosphate

2-[ethyl(propyl)amino]ethyl trihydrogen diphosphate

3,4-Epoxy-1-butenyl diphosphate

-

active-site directed irreversible inhibitor

2958

3,4-epoxy-3-methyl-1-butyl diphosphate

-

653840

3,4-epoxy-3-methylbutyl diphosphate

3,4-oxido-3-methyl-1-butyl diphosphate

-

inactivation through formation of covalent adducts with the reduced flavin, modification of the isoalloxazine ring at position N5

3-(Fluoromethyl)-3-buten-1-yl diphosphate

3-Bromo-3-butenyl diphosphate

-

competitive

2939

3-butyn-1-yl diphosphate

3-cyclopropyl-3-buten-1-yl diphosphate

3-cyclopropylbut-3-en-1-yl trihydrogen diphosphate

-

cyclopropyl-derivative of isopentenyl diphosphate, mechanism-based inhibitor

680372

3-Methyl-3,4-epoxybutyl diphosphate

3-methyl-5-oxo-hexyl diphosphate

-

3-methyl-5-oxo-hexyl diphosphate ethylene glycol acetal

-

3-methylene-4-penten-1-yl diphosphate

3-oxiran-2-ylbut-3-en-1-yl trihydrogen diphosphate

-

epoxy-derivative of isopentenyl diphosphate, mechanism-based inhibitor. The enzyme catalyzes formation of a stable covalent adduct between the inhibitor and FMNH2 in a reaction catalyzed by protonation of the epoxide moiety followed by nucleophilic addition of the cofactor

680372

3-oxiranyl-3-buten-1-yl diphosphate

4-aminophenyl-2-ethane-1,1-bisphosphonic acid

-

weak. Due to inhibition of more than one enzyme in the mevalonate pathway may lead to an increase in antiresorptive potency compared to bisphosphonates that only inhibit farnesyl diphosphate synthase

649588

5-deaza-FMN

-

ATP

-

partial

beta-Diethylaminoethyldiphenylpropylacetate hydrochloride

-

i.e. SKF-525A

bromohydrin

-

651594

dimethylallyl diphosphate

Dimethylallyl diphosphate containing fluorine, epoxy, and ammonium function groups

Claviceps purpurea

-

2947

-

dimethylammonium diphosphate

diphosphate

EDTA

addditon of 5mM EDTA results in almost complete loss of activity

661357

epoxide of isopentenyl diphosphate

-

651594

farnesyl diphosphate

FMN

-

747106, 747530

geranyl diphosphate

geranyl monophosphate

-

homodimethylallyl diphosphate

homoisopentenyl diphosphate

iodoacetamide

7 entries

iodoacetic acid

Isoamyl diphosphate

-

competitive

2939

isopentenyl diphosphate

-

747106, 747530

Isopentenyl diphosphates containing fluorine, epoxy, and ammonium functional groups

Claviceps purpurea

-

2947

-

KH2PO4

-

linaloyl diphosphate

-

linaloyl monophosphate

-

Methyl diphosphate

-

competitive

2939

Mg2+

Cinchona robusta

-

isomerase I is activated by concentrations below 1 mM of Mn2+ or Mg2+, increasing concentrations are inhibitory

Mn2+

N,N-dimethyl-2-amino-1-ethyl diphosphate

-

transition state analogue, competitive

N,N-dimethylaminoethyl diphosphate

-

NEM

-

neryl diphosphate

-

Neryl monophosphate

-

O2

-

the overall reaction is sensitive to O2

678309

Octyl diphosphate

-

p-hydroxymercuribenzoate

-

PCMB

-

Sodium acetate

-

Terpene diphosphate esters

-

-

Terpene monophosphate esters

-

-

additional information

-

not inhibited by 2-[ethyl(propyl)amino]ethyl trihydrogen diphosphate

-

2-mercaptoethanol

dithiothreitol

FMN

-

required as cofactor. The enzyme requires the combined presence of FMN, NADPH and Mg2+

glutathione

-

activates

L-Cysteine hydrochloride

-

activates

mevalonic acid

-

activates

Mevalonic acid lactone

-

weakly activates

NADPH

-

required as cofactor. The enzyme requires the combined presence of FMN, NADPH and Mg2+

0.017

dimethylallyl diphosphate

Cinchona robusta

-

isomerase I

0.0008 - 15.3

isopentenyl diphosphate

70 entries

0.0874 - 0.0904

NADH

additional information

-

substrate binds to the inactive oxidized and active reduced form of enzyme with similar affinities

-

0.000197 - 29900

isopentenyl diphosphate

23 entries

0.031 - 280

isopentenyl diphosphate

8 entries

0.383

(Z)-3-difluoromethyl-2-buten-1-yl diphosphate

-

pH 7.0, 37°C

0.012

(Z)-3-fluoromethyl-2-buten-1-yl diphosphate

-

pH 7.0, 37°C

0.031 - 0.036

2,3-butadien-1-yl diphosphate

0.0565

3,4-epoxy-3-methylbutyl diphosphate

-

pH 7.0, 85°C

678586

0.0486

3,4-oxido-3-methyl-1-butyl diphosphate

-

pH 7.0, 37°C

0.0074

3-(Fluoromethyl)-3-buten-1-yl diphosphate

-

pH 7.0, 37°C

0.048 - 0.049

3-butyn-1-yl diphosphate

0.054

3-cyclopropyl-3-buten-1-yl diphosphate

-

pH 7.0, 37°C

0.008

3-methylene-4-penten-1-yl diphosphate

-

pH 7.0, 37°C

0.0014

3-oxiranyl-3-buten-1-yl diphosphate

-

pH 7.0, 37°C

0.00013

5-deaza-FMN

-

0.0064

FMN

0.292

isopentenyl diphosphate

0.158

(2Z)-3-methylhept-2-en-1-yl trihydrogen diphosphate

Choristoneura fumiferana

-

in 25 mM Tris-HCl, pH 7.0, at 37°C

0.156

(2Z)-3-methylhex-2-en-1-yl trihydrogen diphosphate

Choristoneura fumiferana

-

in 25 mM Tris-HCl, pH 7.0, at 37°C

0.234

(2Z)-3-methyloct-2-en-1-yl trihydrogen diphosphate

Choristoneura fumiferana

-

in 25 mM Tris-HCl, pH 7.0, at 37°C

0.278

(E/Z)-3-cyclopropyl-2-buten-1-yl diphosphate

Thermus thermophilus

-

25°C, pH 8.6

0.003 - 0.0055

2-[ethyl(methyl)amino]ethyl trihydrogen diphosphate

0.007

2-[ethyl(propyl)amino]ethyl trihydrogen diphosphate

Choristoneura fumiferana

-

in 25 mM Tris-HCl, pH 7.0, at 37°C

0.1

3-cyclopropyl-3-buten-1-yl diphosphate

Thermus thermophilus

-

25°C, pH 8.6

0.00114

3-oxiranyl-3-buten-1-yl diphosphate

Thermus thermophilus

-

pH 7.0, 37°C

0.092

dimethylallyl diphosphate

Choristoneura fumiferana

-

in 25 mM Tris-HCl, pH 7.0, at 37°C

0.0008

dimethylammonium diphosphate

Sus scrofa

-

in 50 mM Tris-HCl (pH 7.0), at 37°C

0.131

homodimethylallyl diphosphate

Choristoneura fumiferana

-

in 25 mM Tris-HCl, pH 7.0, at 37°C

0.066

homoisopentenyl diphosphate

Choristoneura fumiferana

-

in 25 mM Tris-HCl, pH 7.0, at 37°C

0.001

-

pyruvate, C13-labeled at positions 2 and 3, aerobic, pH 8, 37°C

0.046

Cinchona robusta

-

0.08

-

dimethylallyl diphosphate, C13-labeled at positions 3, 4 and 5, aerobic, pH 8, 37°C

0.109

-

0.19

-

dimethylallyl diphosphate, anaerobic, pH 8, 37°C

0.23

-

dimethylallyl diphosphate, aerobic, pH 8, 37°C

0.62

-

isopentenyl diphosphate, anaerobic, pH 8, 37°C

0.63

-

isopentenyl diphosphate, aerobic, pH 8, 37°C

1.2

-

pH 8.0

3.6

-

5.92

-

52

-

isoenzyme type II, isopentenyl diphosphate, pH 7.0, 37 °C

73

-

200

-

3000

-

additional information

7 entries

5.5 - 9.3

-

6 - 8.5

-

in presence of Mg2+. A sharper maximum is observed in presence of Mn2+

Narcissus pseudonarcissus

6 - 8

-

2946

6

-

assay at

6.3

-

6.5 - 7

-

6.5

-

6.6

-

7 - 7.8

-

isomerase I

2942

7 - 8

Bombyx mori

-

isomerase II

2942

7

7 - 7.2

-

7

-

7.4

-

4.5 - 9.5

-

pH 4.5: about 39% of maximal activity, pH 9.5: about 60% of maximal activity

5.5 - 7.5

-

pH 5.5: about 35% of maximal activity, pH 7.5: about 30% of maximal activity

7 - 8.5

Cinchona robusta

-

pH 7.0-7.8: maximal activity, pH 8.5: 20% of maximal activity, isomerase I

35

-

37

-

60

-

assay at

719740, 728667

70

-

80

-

maximum activity

85 - 95

-

15 - 55

-

50 - 90

-

5.06

A9LRT7

theoretical

5.8

calculated

5.9

6

-

calculated from amino acid sequence

6.1

6.4

-

calculated from amino acid sequence

Acholeplasma laidlawii

-

-

-

-

-

-

-

-

Camptotheca acuminata

-

UniProt

-

-

-

-

Cinchona robusta

-

2952, 638703

-

Citrus sp.

-

2935

-

Claviceps purpurea

-

2940, 2947

-

-

2943, 2944

-

Claviceps sp. SD58

-

2943

-

Cucurbita sp.

-

-

-

-

Haematococcus lacustris

-

-

-

UniProt

-

A0A067K3N5

UniProt

-

UniProt

-

-

-

Uniprot

type II isopentenyl diphosphate isomerase

Narcissus pseudonarcissus

-

Morinda citrifolia

-

638703

-

-

-

-

-

-

-

-

-

isoform IDI2

UniProt

-

-

Schizosaccharomyces pombe

-

UniProt

-

-

-

2951

-

Staphylococcus aureus IDI-2

type II enzyme

678309

-

Streptococcus pneumoniae CGSP14

-

747530

UniProt

-

2933, 2935, 2936, 727623

-

strain PCC 6803 ORF sll 1556

-

-

660902, 690701

-

Thermus thermophilus HB 27

4 entries

-

660922

-

Vitis vinifera x Vitis riparia

-

706809

-

Vitis vinifera x Vitis vinifera

-

-

-

bud

-

cell suspension culture

Cinchona robusta

-

-

-

developing

2934

fibroblast

-

in normal fibroblasts and Zellweger fibroblasts, in which the synthesis of cholesterol is impaired

-

-

Citrus sp.

-

-

-

A0A067K3N5

-

747645

skeletal muscle

-

isoform IDI2 is only expressed in skeletal muscle

-

highest expression

in mature and tender leaf, expression is higher than in tubers

691183

additional information

chloroplast

Jatropha curcas

A0A067K3N5

-

A9LRT7

-

-

metabolism

physiological function

Bacillus subtilis (strain 168)

18 entries

Escherichia coli (strain K12)

11 entries

3hyq, download, 3D-view

SCOPe (3hyq)

CATH (3hyq)

Q8ZM82

Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)

3sr7, download, 3D-view

SCOPe (3sr7)

CATH (3sr7)

Q8DUI9

Streptococcus mutans serotype c (strain ATCC 700610 / UA159)

4n02, download, 3D-view

SCOPe (4n02)

CATH (4n02)

Streptococcus pneumoniae (strain CGSP14)

Thermus thermophilus (strain HB27 / ATCC BAA-163 / DSM 7039)

21760

-

x * 21760, selenomethionyl form of the enzyme, mass spectrometry

649155

24500

Schizosaccharomyces pombe

-

x * 24500, SDS-PAGE

26300

x * 26300, SDS-PAGE

26864

-

x * 26864, calculation from nucleotide sequence

2951

27180

A9LRT7

theoretical

706249

28500

Aedes aegypti

-

x * 28500, calculated from amino acid sequence

727624

29000

Cinchona robusta

-

x * 29000, isomerase I, SDS-PAGE

29200

30000

A9LRT7

determined by SDS-PAGE

706249

33350

-

x * 33350, calculation from nucleotide sequence

2958

33500

Capsicum annuum

-

gel filtration

2945

33800

Ipomoea batatas

Q0QYT2

x * 33800, calculated

691183

34000

34390

Gossypium barbadense

-

x * 34390, calculated

691400

35000

4 entries

38460

-

calculated; mass spectrometry

39000

-

SDS-PAGE

39897

-

x * 39897, mass spectrometry

678586

40000

-

x * 40000, SDS-PAGE

713655

41492

-

8 * 41492, calculated

43000

4 * 43000, SDS-PAGE

45000

8 * 45000

48000

-

4 * 48000, SDS-PAGE

50000

-

SDS-PAGE

82500

-

gel filtration

173600

gel filtration

197000

gel filtration

345000

-

gel filtration chromatography

?

homotetramer

monomer

octamer

tetramer

sitting drop vapor diffusion method, ligand-free form of the FMN-bound enzyme form at 2.8 A resolution. The octamer forms a D4 symmetrical open, cage-like structure. The monomers of 45000 Da display a classical TIM barrel fold

P50740

652916

comparison of orthorhombic, monoclinic and trigonal crystal forms, up to 2.2 A resolution. Crystallization of free enzyme and in complex wih transition-state analogue N,N-dimethyl-2-amino-1-ethyl diphosphate

-

690288

crystal structure of free and metal-bound C67A mutant enzyme

-

crystal structure of the isomerase-bromohydrin complex

-

651594

crystallographic investigation of phosphoantigen binding

Q46822

661948

crystals soaked with transition state analogue (N,N-dimethylamino)-1-ethyl diphosphate

-

hanging drop vapor diffusion method, crystal structure of the C67A mutant of isopentenyl diphosphate isomerase complexed with the mechanism-based irreversible inhibitor 3,4-epoxy-3-methyl-1-butyl diphosphate

-

653840

hanging drop vapor diffusion method, crystal structures of complexes with transition state analogue N,N-dimethyl-2-amino-1-ethyl diphosphate and the covalently attached irreversible inhibitors 3,4-epoxy-3-methyl-1-butyl diphosphate at 1.96 A resolution

-

652367

hanging drop vapour diffusion method, selenomethionyl form, crystals display trigonal symmetry, with unit-cell parameters, a = b = 71.3 A, c = 61.7 A, and diffract to 1.45 A resolution

-

649155

of wild-type and mutants Y104A, Y104F

-

enzyme shows a flexible N-terminal alpha-helix covering the active pocket and blocking the entrance. Substrate binding induces conformational change in the active site. A water molecule is the direct proton donor for the substrate

Q13907

681412

native enzyme at 1.7 A and in complex with substrate at 1.9 A resolution. comparison with Escherichia coli enzyme structure

Q13907

681413

sitting drop vapor diffusion method, using 0.6 M calcium acetate and 50 mM HEPES pH 7.5

-

713655

molecular modeling of structure and comparison with structures of Streptococcus pneumoniae and Thermus thermophilus enzymes

Pyrococcus furiosus

A6YQV2

695056

crystallized at 20°C using the hanging-drop vapor diffusion method with a reservoir solution containing 0.1 M Tris-HCl (pH 8.0), 0.2 M sodium citrate, and 30% (vol/vol) polyethylene glycol 400 (PEG 400)

-

719740

the covalent adduct formed between irreversible mechanism based inhibitors, 3-methylene-4-penten-1-yl diphosphate or 3-oxiranyl-3-buten-1-yl diphosphate, and the flavin cofactor are investigated by X-ray crystallography and UV-visible spectroscopy. Both the crystal structures of enzyme binding the flavin-inhibitor adduct and the UV-visible spectra of the adducts indicate that the covalent bond is formed at C4a of flavin rather than at N5. The high-resolution crystal structures of enzyme-substrate complexes and the kinetic studies of new mutants confirm that only the flavin cofactor can catalyze protonation of the substrates and suggest that N5 of flavin is most likely to be involved in proton transfer

-

the crystal structures of the substrate-free enzyme and of the substrate-enzyme complexes, in the oxidized and reduced states, are solved to resolutions between 1.99 and 3.1 A, six distinct types of type 2 IDI crystals are obtained

-

sitting drop vapor diffusion method, using HEPES buffer (100 mM, pH 7.5 with 2 M (NH4)2SO4)

-

-

in complex with diphosphate. The diphosphate moiety is located near the conserved residues H10, R97, H152, Q157, E158, and W219, and the flavin cofactor. The putative active site may stabilize a carbocationic intermediate

-

690998

C113S

-

inactive

C113S/114S

-

inactive

C114S

-

inactive

E169/171Q

-

inactive

E169Q

-

inactive

E171Q

-

inactive

W216F

-

10% activity compared to the wild type enzyme

Y105F

-

115% activity compared to the wild type enzyme

C67A

-

inactive mutant enzyme

E116Q

E87Q

K55A

-

the ratio of maximal velocity to turnover number is 66% of that of the wild-type enzyme

K55R

-

the ratio of maximal velocity to turnover number is 28% of that of the wild-type enzyme

R51K

-

the ratio of maximal velocity to turnover number is 4.2% of that of the wild-type enzyme

R83K

-

the ratio of maximal velocity to turnover number is 104% of that of the wild-type enzyme

W161F

-

the ratio of maximal velocity to turnover number is 0.8% of that of the wild-type enzyme

Y104A

-

0.1% of wild-type activity. Crystallization data. The M2+ metal-binding site is absent in the structure, but Mg2+ is still present and bound to C67, E87, and four water molecules

Y104F

-

0.1% of wild-type activity. Crystallization data. General fold of enzyme is similar to wild-type

D216A

-

mutation of highly conserved residue

E13R/R235E

-

the mutant and the wild-type enzyme show similar Vmax values, while the Km of E13R/R235E is smaller than that of the wild type. The mutant is in the tetrameric state even at a concentration where the wild-type enzyme dominantly forms an octamer

719740

E161A

-

mutation of highly conserved residue

E194A

-

mutation of highly conserved residue

E229A

-

mutation of highly conserved residue

H11A

-

mutation of highly conserved residue

H155A

-

mutation of highly conserved residue

K193A

-

mutation of highly conserved residue

K8A

-

mutation of highly conserved residue

N125A

-

mutation of highly conserved residue

N157A

-

mutation of highly conserved residue

Q160A

-

mutation of highly conserved residue

Q160E

-

10-fold decrease in kcat/Km; the mutant shows a 10fold decrease in kcat/Km

Q160H

-

23-fold decrease in kcat/Km; the mutant shows a 23fold decrease in kcat/Km

Q160K

-

130-fold decrease in kcat/Km; the mutant shows a 130fold decrease in kcat/Km

Q160L

-

28-fold decrease in kcat/Km; the mutant shows a 28fold decrease in kcat/Km

Q160N

-

150-fold decrease in kcat, kcat/Km decreases 66-fold; the mutant shows a 150fold decrease in kcat, although kcat/Km only decreases 66fold

R232A

-

mutation of highly conserved residue

R7A

-

mutation of highly conserved residue

S96A

-

mutation of highly conserved residue

T68A

-

mutation of highly conserved residue

L141H

the mutation enhances the enzyme activity 1.1fold

L141H/W256C

the mutations enhance the enzyme activity 1.65fold

L141H/Y195F

the mutations enhance the enzyme activity 2.03fold

L141H/Y195F/W256C

the mutation enhances the enzyme activity 3.13fold

W256C

the mutation enhances the enzyme activity 0.47fold

Y195F

the mutation enhances the enzyme activity 0.71fold

Y195F/W256C

the mutations enhance the enzyme activity 1.14fold

Q154N

-

active site mutant

715286

N37A

-

the mutant is a monomeric but active enzyme with by 20°C lowered melting temperature and reduced binding affinities of FMN (40fold) and a minimal effect on the kcat value compared to the wild type enzyme

747106

additional information

9 entries

37

-

half-life in dilute solution: 30 min. In more concentrated solutions of the protein or dilute enzyme in the presence of 0.01% bovine serum albumin, little loss of activity after 1 h

48

-

melting temperature of mutant Y104A

55

-

melting temperature of mutant Y104F

69

-

melting temperature of wild-type

70

-

1h, IDI retained 50% of the activity

80

Pyrococcus furiosus

A6YQV2

10 min, more than 90% residual activity

695056

high susceptibility to proteolysis

-

unstable in dilute solutions

-

-15°C, stable for several months

-

-20°C, 0.1 M potassium phosphate buffer, pH 7.0, 2 mM dithiothreitol, stable for several weeks

-

2938

-80°C, 0.05 M Tris/HCl, pH 7.5, containing 0.01 mM leupeptin, 2 mM dithiothreitol, 5% glycerol, 55% of the activity is recovered after 3 months

Cinchona robusta

-

-

13 entries

2 isoforms: I and II

Cinchona robusta

-

by affinity chromatography on Ni-NTA resin

-

705515

Co2+ column chromatography, and gel filtration

Catharanthus roseus

-

728493

cobalt column chromatography

Aedes aegypti

-

727624

DEAE Sepharose column chromatography and Resource Phe column chromatograpyh

-

713655

His-Trap column chromatography

747530

Ni-NTA agarose column chromatography

Populus trichocarpa

-

726676

Ni-NTA agarose resin column chromatography

-

747106

Ni-NTA column chromatography

Artemisia annua

-

748956

nickel-nitrilotriacetic acid affinity chromatography

-

on a HisTrap column, for crystallisation, the enzyme is loaded on a HiLoad 16/60 Superdex 200 column

-

partial

Bombyx mori

-

2942

partial. Dye-ligand and immobilized metal ion interaction chromatography are efficient techniques for the rapid batchwise fractionation, from crude plant extracts, of a series of enzymes of prenyl diphosphate metabolism

Salvia officinalis

-

2954

recombinant enzyme

recombinant protein, enzyme purified under aerobic conditions is inactive until the flavin cofactor is reduced by NADPH or dithionite or photochemically

-

TALON metal affinity resin column chromatography

-

4 entries

expressed in Escherichia coli

expressed in Escherichia coli BL21 cells

expressed in Escherichia coli BL21(DE3) cells

expressed in Escherichia coli BL21(DE3) pLysS cells

Populus trichocarpa

-

726676

expressed in Escherichia coli M15pREP4 cells

-

747106

expressed in Escherichia coli M15[pREP4] cells

747530

expressed in Escherichia coli Rosetta cells

expressed in Nicotiana benthamiana

Artemisia annua

-

748956

expressed in Nicotiana tabacum and Escherichia coli BL-21 (DE3) cells

Jatropha curcas

A0A067K3N5

747645

expressed in Synechocystis sp.

-

748859

expression in Escherichia coli

11 entries

expression in Escherichia coli strain BL21(DE3)

-

expression in Escherichia coli strain JMSB0373a

-

expression in Escherichia coli, His6-tagged type II enzyme

-

for expression in Escherichia coli BL21DE3 cells

-

701815

into a pET-vector for expression in Escherichia coli Bl21DE3 cells

-

into the vector pMD18-T for sequencing, and subsequently into pET-28a+ for expression in Escherichia coli BL21DE3 cells

A9LRT7

706249

overexpressed in Escherichia coli

-

2948

overexpression in Escherichia coli

recombinantly expressed with a polyhistidine tag at its N terminus in Escherichia coli BL21

-

719740

the plasmid pDL11, which contains the genes idi, gps and egsA, is constructed by inserting idi and egsA into pCW3, derived from pCL1920, pDL11 is transformed into Escherichia coli TOP10 cells

-

705515

enzyme expression is increased about 4fold at 4°C, about 2.5fold at 0.1 mM NaCl, about 6fold at 0.15 mM mannitol, and about 7.5fold at 0.03 mM methyl jasmonate

Jatropha curcas

A0A067K3N5

747645

enzyme expression is induced 6 and 24 h after treatment with 30 mM Ag+

Salvia miltiorrhiza

H6VLF2

748714

enzyme purified under aerobic conditions is inactive until the flavin cofactor is reduced by NADPH or dithionite or photochemically

-

reconstitution of apo-enzyme with 5-deaza-FMN results in an inactive enzyme, whereas reconstitution with 1-deaza-FMN leads to full recovery of activity

-

678309

reconstitution of metal-free enzyme with Mg2+, Mn2+, Zn2+, Co2+, Ni2+ or Cd2+ generates active enzyme

-