Information on EC 2.7.1.6 - galactokinase

-

CLB.507001.110

gene name

CLB.510667.120

gene name

GAL1

Gal1p

galactokinase

galactose kinase

GALK

GALK1

GalKSpe4

-

kinase (phosphorylating), galacto-

-

kinase, galacto- (phosphorylating)

-

ATP + alpha-D-galactose = ADP + alpha-D-galactose 1-phosphate

Amino sugar and nucleotide sugar metabolism, Galactose metabolism

phospho group transfer

-

BRENDA

degradation of hexoses

KEGG

MetaCyc

D-galactose degradation I (Leloir pathway), D-galactose degradation V (Leloir pathway), D-galactose detoxification, stachyose degradation

ATP:D-galactose 1-phosphotransferase

Part of the Leloir pathway for galactose metabolism. The enzymes from mammals and from the bacterium Escherichia coli have no activity with N-acetyl-alpha-D-galactosamine [4-6].

9030-53-9

-

2'-dATP + D-galactose

2'-dADP + alpha-D-galactose 1-phosphate

-

-

?

2-amino-2-deoxy-D-galactose + ATP

2-amino-2-deoxy-D-galactose 1-phosphate + ADP

-

-

?

2-deoxy-D-galactose + ATP

2-deoxy-D-galactose 1-phosphate + ADP

-

-

?

3'-dATP + D-galactose

3'-dADP + alpha-D-galactose 1-phosphate

-

-

?

3-deoxy-D-galactose + ATP

?

-

-

?

ADP + alpha-D-galactose 1-phosphate

ATP + alpha-D-galactose

-

-

r

alpha-D-galactose + ATP

alpha-D-galactose 1-phosphate + ADP

-

-

?

ATP + 2-amino-2-deoxy-D-galactose

ADP + 2-amino-2-deoxy-D-galactose 1-phosphate

-

-

?

ATP + 2-deoxy-D-galactose

ADP + 2-deoxy-alpha-D-galactose 1-phosphate

ATP + 2-deoxy-D-galactose

ADP + 2-deoxy-D-galactose 1-phosphate

ATP + 3-deoxy-3-methyl-D-galactose

ADP + 3-deoxy-3-methyl-D-galactose 1-phosphate

-

-

?

ATP + 4-deoxy-D-galactose

ADP + 4-deoxy-alpha-D-galactose 1-phosphate

-

substrate only for mutants Y223F, Y223W

-

?

ATP + 6-deoxy-D-galactose

ADP + 2-amino-2-deoxy-D-galactose 1-phosphate

-

-

?

ATP + 6-deoxy-D-galactose

ADP + 6-deoxy-alpha-D-galactose 1-phosphate

ATP + alpha-D-galactose

ADP + alpha-D-galactose 1-phosphate

ATP + D-galactosamine

ADP + alpha-D-galactosamine 1-phosphate

-

substrate: D-galactosamine, specific activity: 12.5 micromol/min/mg

-

?

ATP + D-galactosamine

ADP + D-galactosamine 1-phosphate

ATP + D-galactose

ADP + alpha-D-galactose 1-phosphate

26 entries

ATP + D-galactose

ADP + D-galactose 1-phosphate

-

-

?

ATP + D-galacturonic acid

ADP + alpha-D-galacturonic acid 1-phosphate

-

substrate: galacturonic acid, specific activity: 0.09 micromol/min/mg

-

?

ATP + D-glucose

ADP + alpha-D-glucose 1-phosphate

ATP + D-talose

ADP + alpha-D-talose 1-phosphate

-

substrate only for mutants Y223F, Y223W

-

?

ATP + galactose

ADP + alpha-D-galactose 1-phosphate

6 entries

ATP + L-altrose

ADP + L-altrose 1-phosphate

-

-

?

ATP + L-glucose

ADP + alpha-L-glucose 1-phosphate

-

substrate only for mutants Y223F, Y223W

-

?

ATP + N-acetyl-D-galactosamine

ADP + N-acetyl-alpha-D-galactosamine 1-phosphate

-

-

?

CTP + D-galactose

CDP + alpha-D-galactose 1-phosphate

-

substrate: CTP, specific activity: 0.551 micromol/min/mg

-

?

D-fucose + ATP

?

-

-

?

D-galactosamine + ATP

?

-

-

?

D-talose + ATP

alpha-D-talose 1-phosphate + ADP

-

-

?

dATP + D-galactose

dADP + alpha-D-galactose 1-phosphate

-

substrate: dATP, specific activity: 138 micromol/min/mg

-

?

dCTP + D-galactose

dCDP + alpha-D-galactose 1-phosphate

-

substrate: dCTP, specific activity: 0.931 micromol/min/mg

-

?

deoxy-ATP + D-galactose

deoxyADP + alpha-D-galactose 1-phosphate

-

-

?

dGTP + D-galactose

dGDP + alpha-D-galactose 1-phosphate

-

substrate: dGTP, specific activity: 31.9 micromol/min/mg

-

?

dTTP + D-galactose

?

-

substrate: dTTP, specific activity: 2.5 micromol/min/mg

-

?

dUTP + D-galactose

dUDP + alpha-D-galactose 1-phosphate

-

substrate: dUTP, specific activity: 1.5 micromol/min/mg

-

?

GTP + D-galactose

GDP + alpha-D-galactose 1-phosphate

-

substrate: GTP, specific activity: 28.5 micromol/min/mg

-

?

ITP + D-galactose

IDP + alpha-D-galactose 1-phosphate

-

substrate: ITP, specific activity: 15.7 micromol/min/mg

-

?

UTP + D-galactose

UDP + alpha-D-galactose 1-phosphate

-

substrate: UTP, specific activity: 0.52 micromol/min/mg

-

?

additional information

?

-

10 entries

ATP + galactose

ADP + alpha-D-galactose 1-phosphate

6 entries

additional information

?

-

ATP

Ca2+

Co2+

Fe2+

Mg2+

Mn2+

Ni2+

-

divalent cation required, 15% of the activation with Mg2+

Zn2+

-

can partially replace for Mg2+

additional information

2-(1,3-benzoxazol-2-ylamino)-4-(4-chloro-1H-pyrazol-3-yl)-4,6,7,8-tetrahydroquinazolin-5(1H)-one

-

inhibitory to isoform Galk1, no inhibition of isoform Galk2 up to 40 microM. Uncompetitive with respect to galactose, compound directly competes with ATP at the binding site, and galactose may be required to bind first. Compound is able to lower galactose 1-phosphate levels in primary fibroblasts of patients suffering Classic Galactosemia without significant effect on viability of cells

737825

3-mercaptoethanol

-

adenosine-5'-[gamma-thio]triphosphate

-

ADP

alpha-D-galactose 1-phosphate

amikacin

-

AMP

-

ATP

isoform GalK2; isoform GalK2

bromo alpha-D-galactopyranoside

-

Cd2+

-

Cu2+

D-galactose 1-phosphate

fluoro alpha-D-galactopyranoside

-

galactose

-

above 1 mM

galactose-1-phosphate

-

genistein

-

gentamycin

-

Hg2+

iodoacetamide

-

0.05 mM, complete inactivation

kanamycin A

-

Mg2+

-

free Mg2+ is a competitive inhibitor of MgATP2-

NaCl

50–500 mM, the enzymatic activity decreases approximately 40%–60%, respectively; 50–500 mM, the enzymatic activity decreases approximately 40%–60%, respectively

NEM

neomycin

-

paromycin

-

PCMB

quercetin

-

tobramycin

-

Zn2+

additional information

-

2-mercaptoethanol

-

5 mM, 2-4fold stimulation

cysteine

-

5 mM, 2-4fold stimulation

dithiothreitol

-

5 mM, 2-4fold stimulation

glutathione

-

5 mM, 2-4fold stimulation

KCl

50-100 mM, about 80% increase in activity; 50-100 mM, about 80% increase in activity

2.9

2-amino-deoxy-D-galactose

-

2.2 - 29.21

2-deoxy-D-galactose

6.4

3-deoxy-3--methyl-D-galactose

-

4.9

6-deoxy-D-galactose

-

0.48 - 1.8

alpha-D-galactose

8 entries

0.0039 - 3.83

ATP

45 entries

1.2

D-galactosamine

-

pH 7.3, 25°C

0.091 - 37.23

D-galactose

38 entries

0.038 - 49.37

D-glucose

8 entries

6.04 - 6.28

L-altrose

0.17

MgATP2-

-

pH 7.8

15.76

N-acetyl-D-galactosamine

-

pH 8.0, 45°C

0.195

2-amino-deoxy-D-galactose

-

0.5 - 4.01

2-deoxy-D-galactose

0.085

3-deoxy-3-methyl-D-galactose

-

0.0483

6-deoxy-D-galactose

-

0.05 - 11

alpha-D-galactose

7 entries

0.4 - 146

ATP

11 entries

0.38 - 146

D-galactose

17 entries

0.007 - 1.1

D-glucose

6 entries

0.36

N-acetyl-D-galactosamine

-

pH 8.0, 45°C

0.0022 - 0.059

2-amino-2-deoxy-D-galactose

0.019 - 0.237

2-deoxy-D-galactose

0.024 - 0.054

3-Deoxy-D-galactose

0.064 - 23

alpha-D-galactose

8 entries

1 - 207

ATP

0.00057 - 0.026

D-fucose

0.013 - 0.615

D-galactosamine

0.07 - 164

D-galactose

0.023

D-glucose

-

pH 8.0, 45°C

0.0052 - 0.14

D-talose

0.022

N-acetyl-D-galactosamine

-

pH 8.0, 45°C

0.2 - 8.5

ADP

28 - 90

alpha-D-galactose 1-phosphate

10.2

AMP

-

0.41

ATP

isoform GalK2, pH 8.6, temperature not specified in the publication

2 - 3.5

D-galactose 1-phosphate

0.0739

genistein

-

25

Mg2+

-

0.0225

quercetin

-

0.0047

2-(1,3-benzoxazol-2-ylamino)-4-(4-chloro-1H-pyrazol-3-yl)-4,6,7,8-tetrahydroquinazolin-5(1H)-one

Homo sapiens

-

isoform Galk1, pH not specified in the publication, temperature not specified in the publication

737825

49.9

amikacin

Rattus norvegicus

-

4.4

bromo alpha-D-galactopyranoside

Rattus norvegicus

-

0.86

fluoro alpha-D-galactopyranoside

Rattus norvegicus

-

16.8

Galactose 1-phosphate

Rattus norvegicus

-

36.6

gentamycin

Rattus norvegicus

-

20

kanamycin A

Rattus norvegicus

-

30

neomycin

Rattus norvegicus

-

67.3

paromycin

Rattus norvegicus

-

7.2

tobramycin

Rattus norvegicus

-

0.0043

-

mutant R36K, synthesis of ATP, pH 8.0, 24°C

0.0067

-

mutant R36A, synthesis of ATP, pH 8.0, 24°C

0.07

-

R37A mutant protein, substrate: galactose

0.0781

-

substrate: ATP + 6-deoxy-D-galactose, pH 8, 37°C

0.08

-

mutant R36A, synthesis of galactose 1-phosphate, pH 8.0, 24°C

0.0812

-

0.09

-

substrate: ATP + D-galacturonic acid, pH 8, 37°C

0.1

-

mutant R36K, synthesis of galactose 1-phosphate, pH 8.0, 24°C

0.14

-

Y109F mutant protein, substrate: galactose

0.15

-

0.17

-

Y236F mutant protein, substrate: galactose

0.43

-

wild-type, synthesis of ATP, pH 8.0, 24°C

0.52

-

substrate: UTP + D-galactose, pH 8, 37°C

0.551

-

substrate: CTP + D-galactose, pH 8, 37°C

0.61

-

mutant Y223H

0.63

-

mutant Y223R

0.65

-

mutant Y223L

0.87

-

mutant Y223W

0.91

-

wild-type protein, substrate: galactose

0.931

-

substrate: dCTP + D-galactose, pH 8, 37°C

1.14

-

Y109H mutant protein, substrate: galactose

1.5

-

substrate: dUTP + D-galactose, pH 8, 37°C

1.62

-

enzyme from erythrocyte

1.737

-

enzyme from placenta

2.258

-

2.5

-

substrate: dTTP + D-galactose, pH 8, 37°C

9.6

-

641632

11.5

-

wild-type, synthesis of galactose 1-phosphate, pH 8.0, 24°C

12.5

-

substrate: ATP + D-galactosamine, pH 8, 37°C

15.7

-

substrate: ITP + D-galactose, pH 8, 37°C

23.42

-

mutant Y223F

23.5

-

wild-type

28.5

-

substrate: GTP + D-galactose, pH 8, 37°C

31.9

-

substrate: dGTP + D-galactose, pH 8, 37°C

38.3

-

641708, 641713

99.7

-

substrate: ATP + 2-deoxy-D-galactose, pH 8, 37°C

137

-

substrate: ATP + D-galactose, pH 8, 37°C

138

-

substrate: dATP + D-galactose, pH 8, 37°C

158

-

substrate: ATP + D-galactose, pH 8, 37°C

318

-

additional information

6

-

6.5 - 7.5

-

7.3

-

7.4

-

7.5 - 8

-

in Tris or triethanolamine buffer

7.5 - 9.5

; isoform GalK1

7.6

-

7.7 - 7.9

-

Tris-glycine-NaH2PO4 buffer

7.8

-

7.8 - 8

-

7.8

-

8

-

assay at

8 - 9

-

641713

8

8.5

-

optimal pH

3 - 11

-

5.5 - 8

-

pH 5.5: about 60% of maximal activity, pH 8.0: about 70% of maximal activity, potassium phosphate buffer

6.7 - 8.7

-

pH 6.7: about 45% of maximal activity, pH 8.7: about 35% of maximal activity

7 - 9

37

-

assay at

41

-

45

55

-

highest activity

20 - 60

-

20°C: about 30% of maximal activity, 60°C: about 40% of maximal activity

5.1

-

isoelectric focusing

5.3

-

isoelectric focusing

5.7

5.8

-

isoelectric focusing

641719

Arabidopsis thaliana

-

723394

-

-

641715, 641718

-

-

-

Canis lupus familiaris

dog breed Labrador retriever

SwissProt

-

-

-

-

galactose mutant

-

Mycobacterium sp. 279

galactose mutant

641706

-

-

641725, 662617

-

-

-

-

-

-

641708, 641713, 641717, 641727, 674492, 674554, 674579

-

-

Saccharomyces cerevisiae W303alpha

-

-

674554

-

-

Streptococcus thermophilus

-

Streptococcus salivarius

expression in Streptococcus thermophilus

660753

-

strains IMDOST40 and IMDOST07

TrEMBL

-

-

-

-

Trichoderma reesei

Trichoderma reesei QM9414

synonym Trichoderma reesei

-

-

-

-

-

expression of both isoforms GalK1 and GalK2; expression of both isoforms GalK1 and GalK2

-

-

-

expression of both isoforms GalK1 and GalK2; expression of both isoforms GalK1 and GalK2

both isoforms GalK1 and GalK2; both isoforms GalK1 and GalK2

20015

malfunction

Arabidopsis thaliana

-

an AtGALK T-DNA insertion mutant (atgalk) that (a) is AtGALK transcript deficient, (b) displays no GALK activity in vegetative tissues; and (c) accumulates Gal up to 6.8 mg/g FW in vegetative tissues, in contrast to wild-type plants. By constitutively overexpressing functional AtGALK cDNA, atgalk is functionally rescued

723394

7 entries

1pie, download, 3D-view

SCOPe (1pie)

CATH (1pie)

Q9R7D7

Lactococcus lactis subsp. lactis (strain IL1403)

1s4e, download, 3D-view

SCOPe (1s4e)

CATH (1s4e)

Q9HHB6

Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1)

Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)

2aj4, download, 3D-view

SCOPe (2aj4)

CATH (2aj4)

P04385

Saccharomyces cerevisiae (strain ATCC 204508 / S288c)

26000

-

2 * 26000, SDS-PAGE

28000

-

2 * 28000, SDS-PAGE

30900

-

2 * 30900, SDS-PAGE

37000 - 38000

-

gel filtration

37000

-

calculatd from cDNA

38000

-

1 * 38000, SDS-PAGE

40000

-

gel filtration

641725

41700

Mycobacterium sp.

-

gel filtration

641706

43000

-

x * 43000 Da, SDS-PAGE

45000

-

SDS-PAGE

46000

-

gel filtration, calculation from sedimentation and diffusion data

47000

-

gel filtration

48000

-

1 * 48000, SDS-PAGE

50000 - 55000

-

gel filtration

50000

gel filtration, both isoforms GalK1 and GalK2; gel filtration, both isoforms GalK1 and GalK2

51300

-

51900

-

53000 - 57000

-

gel filtration

641712

55000

-

gel filtration

57000

Trichoderma reesei

-

x * 57000, calculated

662892

58000

60000

additional information

?

dimer

monomer

-

-

in complex with D-galactose and adenosine 5’-(beta,gamma-imino)-triphosphate

-

662453

hanging drop method of vapor diffusion, wild-type and selenomethionine-labeled galactokinase

-

641724

hanging-drop method of vapour diffusion with ammonium sulfate as the precipitant. The crystals diffract X-rays to 2.9 A resolution on a synchroton-radiation source

-

641725

in complex with MgADP and D-galactose, comparison with S. cerevisiae enzymes

-

662617

seeding method in combination with the sitting-drop vapour-diffusion method using 30%(v/v) PEG 400, 1 M D-galactose, 5 mM AMP-PNP, 5% (w/v) PEG 4000, 50 mM MgCl2 and 0.1 M MES pH 6.3.

-

hanging drop vapour diffusion method using 18-25% poly(ethylene glycol) 8000 containing 200 mM LiCl buffered with 100 mM MES3 (pH 6.0) and enzyme concentrations of 15.0 mg/ml and 5 mM Mg-adenosine 5'-(beta, gamma-imido)triphosphate

-

674492

M173L

-

moderate activity on D-glucose, wider substrate specificity than wild-type

M173L/Y371H

-

acts on additional substrates such as 4-azido-4-deoxy-D-galactose, 6-ido-6-deoxy-D-galactose, 6-chloro-6-deoxy-D-galactose, 6-bromo-6-deoxy-D-galactose, 4-deoxy-D-galactose, 6-thio-6-deoxy-D-galactose, 6-thio-6-deoxy-D-glucose, 6-azido-6-deoxy-D-glucose

Y223F

-

activity similar to wild-type, converts additional substrates L-glucose, D-talose, 4-deoxy-D-galactose, less active on D-galactosamine than wild-type

Y223H

-

significant reduction in activity

Y223L

-

significant reduction in activity

Y223R

-

significant reduction in activity

Y223W

-

significant reduction in activity, converts additional substrates L-glucose, D-talose, 4-deoxy-D-galactose, with less efficiency than mutant Y223F, less active on D-galactosamine than wild-type

Y371H

-

wider substrate specificity than wild-type

A198V

A384P

-

mutant enzyme is not present in the soluble fraction after sonication and can not be purified

C32M

-

mutant enzyme is not present in the soluble fraction after sonication and can not be purified

D46A

-

no detectable activity

E43A

-

minor changes in kinetic parameters

E43G

-

10fold drop in turnover number

E43G/H44I

-

not soluble

G346S

-

mutant enzyme shows substantial reduction in turnover number. Lower specificity constant for galactose than wild-type enzyme

G347S

-

mutant enzyme shows substantial reduction in turnover number, increase in Km-value for galactose. Lower specificity constant for galactose than wild-type enzyme

G349S

G36R

-

mutant enzyme is not present in the soluble fraction after sonication and can not be purified

H44A

-

not soluble

H44I

-

not soluble

H44Y

-

increrase in Km-value for galactose compared to wild-type. Lower specificity constant for galactose than wild-type enzyme

P28T

-

mutant enzyme is not present in the soluble fraction after sonication and can not be purified

R256W

-

drastic reduction of activity when expressed in COS cells, missense mutation causes GALK deficiency

641721

T288M

-

mutant enzyme is not present in the soluble fraction after sonication and can not be purified

T344M

-

drastic reduction of activity when expressed in COS cells, missense mutation causes GALK deficiency

641721

D183A

-

inactive

D183N

-

inactive

R36A

-

100fold decrease in activity

R36N

-

100fold decrease in activity

D186A

-

residue D186 interacts with galactose, possible catalytic residue, no activity of mutant enzyme

D186E

-

residue D186 interacts with galactose, possible catalytic residue, no activity of mutant enzyme

D186N

-

residue D186 interacts with galactose, possible catalytic residue, no activity of mutant enzyme

R37A

-

residue R37 interacts with galactose

Y109F

-

residue Y109interacts with ATP

Y109H

-

residue Y109interacts with ATP

Y236A

-

residue Y236 interacts with galactose

Y236F

-

residue Y236 interacts with galactose

D62A

-

abolishes all detectable galactokinase activity but retains the ability to use D-glucose as a substrate

D62A/Y274A

-

shows no sugar kinase activity

D62C

-

shows no sugar kinase activity

D62E

-

shows no detectable sugar kinase activity

D62F

-

abolishes all detectable galactokinase activity but retains the ability to use D-glucose as a substrate

D62G

-

shows no sugar kinase activity

D62H

-

abolishes all detectable galactokinase activity but retains the ability to use D-glucose as a substrate

D62I

-

shows no sugar kinase activity

D62K

-

shows no detectable sugar kinase activity

D62L

-

abolishes all detectable galactokinase activity but retains the ability to use D-glucose as a substrate

D62M

-

shows no sugar kinase activity

D62N

-

shows no detectable sugar kinase activity

D62P

-

shows no sugar kinase activity

D62Q

-

shows no sugar kinase activity

D62R

-

shows no sugar kinase activity

D62S

-

shows no sugar kinase activity

D62T

-

shows no sugar kinase activity

D62V

-

shows no sugar kinase activity

D62W

-

shows no sugar kinase activity

D62Y

-

shows no sugar kinase activity

R53A/D217A

-

shows no sugar kinase activity

D62A

-

abolishes all detectable galactokinase activity but retains the ability to use D-glucose as a substrate

-

D62E

-

shows no detectable sugar kinase activity

-

D62G

-

shows no sugar kinase activity

-

D62M

-

shows no sugar kinase activity

-

D62N

-

shows no detectable sugar kinase activity

-

additional information

4

-

very labile below

641715

4.5

-

37°C, 10 min, about 20% loss of activity

641715

4.5 - 6.5

-

25°C, 30 min, stable

641711

5.4 - 8.8

-

24 h, less than 20% loss of activity

641708

6

-

stable above

641715

7

Mycobacterium sp.

-

stable

641706

4 - 30

-

only 20% of maximal activity

20 - 45

-

pH 7.3, 50 mM phosphate buffer, 10 mM 2-mercaptoethanol, 5% glycerol, 60 min, stable

30

-

10 min, no loss of activity

641719

37

-

very little activity at 37°C

40

-

pH 7.5, 10 min, stable up to

50

55

-

nearly inactive at 55°C

60

65

-

over 76% of maximal activity

70

-

10 min, complete loss of activity without dithiothreitol

90

95

-

20 min, 2 mM dithiothreitol, 60% loss of activity

dithiothreitol protects the enzyme better than 2-mercaptoethanol against denaturation on storage at 4°C

-

641719

overnight dialysis at pH above 9.2 or below pH 5.5 results in loss of all enzymatic activity

-

641713

the enzyme is stabilized during the purification procedure by the inclusion of 10% glycerol, 1 mM phosphate and 20 mM 2-mercaptoethanol in all buffer solutions

Cricetulus griseus

-

641707

-20°C, 50% loss of activity after 1 months

-

-20°C, unstable in 0.01 M KH2PO4-K2HPO4, pH 7.2 buffer, 0.01 M MgCl2, 1 mM dithiothreitol, 0.2 mM ATP without glycerol

-

-30°C, stable for 2 months with only 5-10% loss of activity

-

-80°C, less than 20% loss of activity after 6 months

-

641708, 641713

0°C, galactokinase suspension in saturated (NH4)2SO4 retains at least 80% of its activity after 1 week

-

4°C, 0.01 M KH2PO4-K2HPO4, pH 7.2 buffer, 0.01 M MgCl2, 1 mM dithiothreitol, about 40% loss of activity after 24 h

-

4°C, 24 h, gradual loss of activity within 2 weeks

-

641632

4°C, 3.0 M ammonium sulfate, 10% loss of activity after 6 d

-

4°C, potassium phosphate buffer, pH 7.5, more than 80% of the initial activity remains after 1 week

-

-

10 entries

HiTrap Q column chromatography, HiTrap Phenyl FF column chromatography, and HiLoad 16/60 Superdex 75 gel filtration

-

Ni2+-nitrilotriacetic acid-agarose column chromatography

-

nickel-nitrilotriacetic acid-agarose chromatography and Resource-Q column chromatography

-

674492

one-step purification, affinity chromatography

-

partial

-

recombinant enzyme

-

two forms of galactokinase, one of which is purified to homogeneity

Cricetulus griseus

-

641707

using Ni-NTA chromatography

-

-

expressed in Escherichia coli

expressed in Escherichia coli DH5alpha cells

-

674492

expressed in Escherichia coli strain BL21 codon plus (DE3)-RIL

-

expressed in Escherichia coli strains JM109 and K12

Trichoderma reesei

-

674821

expression in COS cells, galactokinase maps to the distal region of mouse chromosome 11

Mus musculus

-

641722

expression in COS7 cells

P51570

689012

expression in Escherichia coli

expression in Escherichia coli; expression in Escherichia coli

gene from Escherichia coli K-12 is subcloned into pET-15b vector and overexpressed in BL21 cells

-

His-tagged, expressed in Escherichia coli BL21(DE3)

-

mutant and wild-type cDNA introduced into COS cells

-

641721

overexpression in Escherichia coli

-

2328

enhanced gene expression caused by a nucleotide variation within GALKI promoter (c.-22T>C mutation)

-

702897

medicine

nutrition

Streptococcus salivarius

-

expression of enzyme in Streptococcus thermophilus, recombinant strain grows on galactose with a generation time of 55 min, which is almost double the generation time on lactose. During growth on milk and under conditions simulating those used to produce mozzarella cheese, recombinant strain grows and produces acid more rapidly than wild-type strain

660753

synthesis

-

enzyme mutant Y371H/M173L, substrate range is much wider than wild-type, generation of unnatural sugar 1-phosphates