Information on EC 3.5.99.6 - glucosamine-6-phosphate deaminase

-

Substrates: -
Products: -

r

D-fructose 6-phosphate + NH3

D-glucosamine 6-phosphate + H2O

-

Substrates: deamination reaction is preferred over amination/isomerization
Products: -

r

D-glucosamine 6-phosphate + H2O

D-fructose 6-phosphate + NH3

32 entries

glucosamine 6-phosphate + H2O

fructose 6-phosphate + NH3

6 entries

additional information

?

-

9 entries

alpha-D-glucosamine 6-phosphate + H2O

D-fructose 6-phosphate + NH3

15 entries

D-fructose 6-phosphate + NH3

alpha-D-glucosamine 6-phosphate + H2O

-

Substrates: -
Products: -

r

D-glucosamine 6-phosphate + H2O

D-fructose 6-phosphate + NH3

glucosamine 6-phosphate + H2O

fructose 6-phosphate + NH3

additional information

?

-

6 entries

arsenate

-

activates

Co2+

-

activates

Hg2+

-

activates

Mg2+

-

activates

Mn2+

-

activates, reverses inhibition by EDTA

Ni2+

-

activates

Zn2+

required, the sigmoidal response with respect to Zn2+ concentration suggests cooperativity and the binding of at least two metal ions within the active site

additional information

2,5-anhydro mannitol 6-phosphate

-

2-deoxy-2-amino-D-glucitol 6-phosphate

8 entries

5,5'-dithiobis(2-nitrobenzoate)

-

648227

alpha-D-glucosamine 6-phosphate

substrate inhibition

Cd2+

-

Cu2+

-

cysteine

-

D-fructose 1,6-diphosphate

-

D-fructose 6-phosphate

D-fructose oxime 6-phosphate

-

D-glucosamine 6-phosphate

-

substrate inhibition

669338

D-glucose 6-phosphate

-

diethyl dicarbonate

complete inactivation, 2-deoxy-2-amino-D-glucitol 6-phosphate protects

diphosphate

-

EDTA

fluorescamine

-

N-acetylglucosamine 6-phosphate

-

N3-(4-Methoxyfumaroyl)-L-2,3-diaminopropanoic acid

-

Ni2+

-

PCMB

phosphate

-

slight inhibition

UDP-N-acetylglucosamine

-

slight inhibition at 1.67 mM, slight activation at 0.00167-0.167 mM

Zn2+

additional information

-

2-deoxyglucose 6-phosphate

-

activates

6-phosphogluconate

-

activates

D-glucose 6-phosphate

D-mannose 6-phosphate

-

GlcNAc6P

enzyme NagB is allosterically activated by GlcNAc6P

757097

glucosamine-6-phosphate

Shewanella denitrificans

homotropic activation

N-acetylgalactosamine

-

N-Acetylgalactosamine 6-phosphate

Sus scrofa

-

activates

N-acetylglucosamine

-

N-acetylglucosamine 6-phosphate

14 entries

N-acetylglucosamine-6-phosphate

-

allosteric activator of NagB

712289

PCMB

-

inhibits at high concentrations, activates at low concentrations

UDP-N-acetylglucosamine

-

slight activation at 0.00167-0.167 mM, slight inhibition at 1.67 mM

additional information

5 entries

-

0.24 - 45.2

alpha-D-glucosamine 6-phosphate

15 entries

1.1 - 36

D-fructose 6-phosphate

14 entries

0.041 - 17

D-glucosamine 6-phosphate

51 entries

1.7

fructose 6-phosphate

-

amination reaction, pH 8.0, 30°C

5

glucosamine 6-phosphate

-

deamination reaction, pH 8.0, 30°C

2 - 41.6

NH3

3.7 - 300

NH4+

10 entries

additional information

15 entries

-

105 - 220

alpha-D-glucosamine 6-phosphate

5 entries

5.8 - 455

D-fructose 6-phosphate

0.044 - 1800

D-glucosamine 6-phosphate

36 entries

39.6

NH3

-

pH 7.3, 60°C

additional information

5 entries

-

73.3 - 437.5

alpha-D-glucosamine 6-phosphate

13.4

2,5-anhydro mannitol 6-phosphate

pH 7.7, 30°C

0.0041

2-deoxy-2-amino D-glucitol 6-phosphate

-

pH 7.7, 30°C

0.002 - 0.0043

2-deoxy-2-amino-D-glucitol 6-phosphate

9 entries

3

alpha-D-glucosamine 6-phosphate

pH 8.0, 20°C, recombinant enzyme

1.3

D-fructose oxime 6-phosphate

pH 7.7, 30°C

4

D-glucosamine 6-phosphate

-

22°C, pH 8.0

669338

0.036

-

initial culture, cell homogenate

0.5

-

after 7.2fold purification, HisC-tagged enzyme, with D-fructose 6-phosphate as substrate, at pH 7.0 and 37°C

158

-

27.6

-

308

-

52

-

606.8

-

62

-

after 7.2fold purification, untagged wild type enzyme, with D-fructose 6-phosphate as substrate, at pH 7.0 and 37°C

73

-

after 7.2fold purification, HisN-tagged enzyme, with D-fructose 6-phosphate as substrate, at pH 7.0 and 37°C

8.6

-

crude extract, with D-fructose 6-phosphate as substrate, at pH 7.0 and 37°C

85

-

purified enzyme

additional information

6.5 - 8

-

6.6

-

in absence of activator

7

-

7 - 8.2

-

7.1 - 7.3

-

in absence of activators

7.2

-

7.4

7.5

-

enzyme activated by N-acetylgalactosamine 6-phosphate

7.5 - 8

-

7.8

-

7.8 - 8.2

Oryctolagus cuniculus

-

648218

8

8 - 8.5

-

both amination and deamination reaction

8.2 - 8.4

-

in presence of phosphate and D-glucose 6-phosphate

8.3

-

8.5 - 8.7

-

8.8

8.9

-

both directions

648228

9

-

6 - 10

-

pH-profile, overview

6 - 8.5

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pH 6.0: about 25% of maximal activity, pH 8.5: about 60% of maximal activity

6 - 9

the pH dependency of NagB with a standard concentration of GlcNAc-6P, the kcat does not vary between pH 6.0 and pH 9.0. The Hill coefficient changes substantially with an optimum at pH 7.7 (Hill coefficient of 2.6), dropping below pH 7.0 or above pH 8.8. Results obtained at different pH values (pH 8.0, 7.8, 7.5, and 6.8) are in agreement with this statement

757101

6.2 - 8

Drosophila virilis

-

pH 6.2: about 25% of maximal activity, pH 8.0: about 55% of maximal activity, in presence of glucose 6-phosphate

648222

6.8 - 8.3

-

70% of maximal activity at pH 6.8, maximal activity at pH 8.3

7 - 9.5

-

pH 7.0: about 55% of maximal activity, pH 9.5: about 74% of maximal activity

additional information

20

assay at

30

37

95 - 100

-

32 - 47

-

about 90% of maximal activity at 32°C and 47°C

additional information

-

effect of temperature on homotropic co-operativity in the enzyme

648243

5.7

-

isoelectric focusing and chromatofocusing

5.9

-

electrofocusing on polyacrylamide gels

8.55

-

chromatofocusing

-

-

Aspergillus niger A158

-

-

Bacillus subtilis 168

-

UniProt

-

-

-

-

Candida albicans BWP17

-

710931

-

Haemophilus influenzae 86-028NP

-

-

-

-

-

-

-

-

-

-

Haemophilus influenzae

-

779120

Q4QP46

SwissProt

-

SwissProt

-

-

-

-

-

-

Oryctolagus cuniculus

-

648218

-

Pasteurella multocida

-

779120

Q9CMF4

SwissProt

Pasteurella multocida Pm70

-

SwissProt

-

-

-

-

-

-

Shewanella denitrificans

cf. EC 2.6.1.16

Shewanella denitrificans OS217

UniProt

cf. EC 2.6.1.16

UniProt

Shewanella oneidensis

-

Shewanella oneidensis ATCC 700550

Q8EBK7

UniProt

-

Q8EBK7

UniProt

-

UniProt

Staphylococcus aureus TCH1516

-

Staphylococcus aureus USA300

UniProt

-

UniProt

Streptococcus mutans UA159

gene smu.636

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Sus scrofa

hog

-

-

-

adipose tissue

subcutaneous fat and abdominal fat, high expression level

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low expression level

duodenum

high expression level

low expression level

glandular stomach

low expression level

hypophysis

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hypothalamus

high expression level

-

-

-

low expression level

preadipocyte

-

-

-

-

-

-

telencephalon

low expression level

-

-

additional information

10 entries

Highest Expressing Human Cell Lines

Filter by:

Cell Line Links

Gene Links

evolution

6 entries

malfunction

metabolism

physiological function

Borreliella burgdorferi B31

additional information

Escherichia coli K-12

Haemophilus influenzae 86-028NP

Klebsiella pneumoniae

Pasteurella multocida subsp. multocida str. Pm70

Vibrio cholerae O1 biovar El Tor str. N16961

154000

-

sucrose density gradient centrifugation

17500

Candida albicans

-

gel filtration

648224

180000

-

about, gel filtration

185000

-

gel filtration

648226

190000

-

pore-gradient gel electrophoresis, density gradient centrifugation

648226

210000

-

gel filtration

25500

Streptococcus mutans

-

x * 25500, recombinant His-tagged enzyme, SDS-PAGE

28500

monomeric enzyme, gel filtration

29000

-

gel filtration

648228

29700

30400

-

6 * 30400, SDS-PAGE

30700

-

x * 30700, native enzyme, SDS-PAGE

32500

-

1 * 32500, SDS-PAGE

33000

Mus musculus

-

x * 33000, SDS-PAGE

648242

37300

-

2 * 37300, SDS-PAGE

57000

dimeric enzyme, gel filtration

71800

-

gel filtration

?

dimer

hexamer

homohexamer

monomer

monomer or dimer

monomer or homodimer or homotetramer

-

the recombinant and wild type enzyme exhibit heterogeneity of the quaternary structure and in solution exist in three interconvertible forms, namely, monomeric, homodimeric, and homotetrameric. The monomer/dimer/tetramer ratios depended on protein concentration and fall within the range from 72:27:1 to 39:23:38. The enzyme is fully active in each of the oligomeric structures

additional information

side-chain modification

-

-

648235

structure of the enzyme in complexes with its allosteric activator and with a competitive inhibitor

-

648229

the crystallographic structure of F174A is determinated

-

648244

the structure of five complexes of the enzyme in R and T state is described and their kinetic and allosteric implication analysed, the ligand-free enzyme, T-conformer, undergoes an allosteric transition to the free active-site R conformer upon binding of the allosteric activator

648240

hanging-drop vapor-diffusion method at 18°C, crystallized in the P2(1) space group with 12 monomers in the asymmetric unit. The structure of HiNagB is determined to 3.0 A resolution

Haemophilus influenzae

-

isozyme GNPDA1, X-ray diffraction structure determination and analysis at 1.75 A resolution

P46926, Q8TDQ7

718986

hanging-drop vapor-diffusion method at 18°C, crystallized in the P2(1) space group with 4 monomers in the asymmetric unit. The structure of PmNagB is determined to 2.3 A resolution

Pasteurella multocida

Q9CMF4

779120

sitting-drop vapor diffusion method, hanging drop method at 18°C. The enzyme crystallized in two different space groups, C222(1) in the presence of GlcNol6P and P2(1)2(1)2(1) for the GlcNol6P-GlcNAc6P complex and in the absence of ligands

Shewanella denitrificans

purified recombinant His-tagged enzyme, the protein solution contains 10 mg/ml protein in 20 mM Tris-HCl, 150 mM NaCl, pH 7.5, mixing of 0.001 ml of protein and 0.001 ml of reservoir solution, and equilibration against 0.5 ml of reservoir solution, hanging drop vapour diffusion method, X-ray diffraction structure determination and anaylsis at 2.4 A resolution

-

C118S/C228S/C239S

decrease in kcat-value, no modification of allosteric activation

667638

C118Ser/C228S/C239S/D165C/S206W

the mutant shows reduced turnover number compared to the wild type enzyme

735143

C219S

-

the kinetic and allosteric properties of the mutant enzyme in which Ser replaces Cys219 or Cys228 are the same as those described for the wild-type enzyme. The same result is obtained with the double mutation

648230

C228S

-

the kinetic and allosteric properties of the mutant enzyme in which Ser replaces Cys219 or Cys228 are the same as those described for the wild-type enzyme. The same result is obtained with the double mutation

648231

D141N

the mutation modifies the kcat versus pH profile of the enzyme

D141N/E148Q

mutation modifies the kcat versus pH profile of the enzyme

E148Q

the mutation modifies the kcat versus pH profile of the enzyme

F174A

H143Q

drastically impairs the activity of the enzyme in the forward but not in the backward direction of the reaction

K208E

entirely inactive in absence of allosteric activator N-acetylglucose-6-phosphate

K208V

homotropic cooperativity, Hill-coefficient 1.7, 2fold increase in dissociation constant for allosteric activator N-acetylglucose-6-phosphate compared to wild-type

R172A

inactive in absence of allosteric activator N-acetylglucose-6-phosphate, at high activator levels, cooperativity diminishes and substrate inhibition becomes significant

R172A/K208E

inactive in absence of allosteric activator N-acetylglucose-6-phosphate, at high activator levels, cooperativity diminishes and substrate inhibition becomes significant

W15Y

mutant containing a single Trp residue at W224

W15Y/F174W/W224Y

mutant containing a single, new Trp-residue at F174W

W15Y/W224Y

mutant without Trp residues

W15Y/W224Y/Y254W

mutant containing a single, new Trp-residue at Y254W

W224Y

mutant containing a single Trp residue at W15

Y121T

-

while the wild-type enzyme behaves as a classical allosteric K-system which can be described by the allosteric concerted model, the mutant forms Y121T and Y121W present an asymmetric behaviour towards the allosteric activator, which can be described as two distinct half-of-the-sites allosteric activation steps occuring with different affinities for the N-acetyl-D-glucosamine 6-phosphate

648231

Y121W

-

while the wild-type enzyme behaves as a classical allosteric K-system which can be described by the allosteric concerted model, the mutant forms Y121T and Y121W present an asymmetric behaviour towards the allosteric activator, which can be described as two distinct half-of-the-sites allosteric activation steps occuring with different affinities for the N-acetyl-D-glucosamine 6-phosphate

648231

Y254F

-

the mutant is less active than wild-type enzyme, the replacement causes an uncoupling of the homotropic and heterotrophic effects

648237

Y254T

-

the mutation results in pure K-system with a similar catalytic activity to that of the wild-type enzyme, mutant displays kcat values similar to the wild-type enzyme

648237

additional information

25 entries

5 - 9

-

4°C, purified enzyme, overnight, in presence of different salts, stable

60

65

Rattus norvegicus

-

complete loss of activity

648223

80

-

half-life 103 min

90

-

half-life 19 min

D-glucosamine 6-phosphate and N-acetylglucosamine 6-phosphate stabilize against heat inactivation at 60°C, and protect the enzyme from tryptic digestion

Rattus norvegicus

-

648223

introduction of the oligoHis tag at the enzyme's C-terminus results in almost complete loss of the catalytic activity, while the catalytic properties of HisN-tagged enzyme and the untagged wild type are very similar

-

labile to freezing and thawing

protease-sensitive. Incubation with 0.3 mg/ml protease type V or 0.3 mg/ml trypsin inactivates completely

-

stable to dialysis

-

-18°C, little loss of activity after 2-3 months

-

-20°C, 50% glycerol, stable for up to 2 months

-

648228

-20°C, in 150 mM Tris-HCl buffer, pH 7.25, 50% v/v glycerol, stable up to 6 months

-

648226

-70°C, stable for 70 days

Rattus norvegicus

-

648223

0-5°C, in liquid N2, little loss of activity after 2-5 months

-

648221

0°C, stable for at least 2 weeks

Candida albicans

-

648224

4°C, purified enzyme, pH 5.0-9.0, overnight, in presence of different salts, stable

-

-

14 entries

ammonium sulfate precipitation, Resource Q column chromatography, and Ni2+-IDA agarose column chromatography

-

N-6-aminohexanoyl-glucosamine 6-phosphate agarose column chromatography

735143

native enzyme 1390fold from kidney cortex by affinity chromatography on N-epsilon-amino-n-hexanoyl-D-glucosamine-6-phosphate agarose and on phosphocellulose, the latter behaving as an active site affinity resin

-

native enzyme by ammonium sulfate fractionation, anion exchange chromatography, and affinity chromatography on a N-aminohexyl-glucosamine-6-phosphate resin folowed by dialysis and ultrafiltration

-

recombinant enzyme

recombinant enzyme NagB from Escherichia coli strain BL21(DE3) by anion exchange chromatography, hydrophobic interaction chromatography, and gel filtration, to near homogeneity

recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) to homogeneity in two steps by nickel affinity chromatography and gel filtration

Streptococcus mutans

-

recombinant isozyme GNPDA1 from Escherichia coli strain IBPC 590 by single purification step by means of allosteric-site affinity chromatography on N-6-aminohexanoyl-GlcN6P agarose

P46926, Q8TDQ7

718986

wild-type enzyme and two mutants

-

-

-

expressed in Escherichia coli BL21(DE3) pLysS cells

-

expressed in Escherichia coli strain LAA20

-

712289

expressed in Saccharomyces cerevisiae strain BY

Candida albicans

-

710931

expression in Escherichia coli

expression in Escherichia coli BL21(DE3)

expression of isozyme GNPDA1 in Escherichia coli strain IBPC 590 (DELTAnagEBACD DELTAlac), which carries a deletion of the chromosomal copy of the nag operon and expresses the recombinant enzyme constitutively from the lac promoter due to the lacI mutation

P46926, Q8TDQ7

718986

expression of native GND, but not a cytosolic form of GND, in DELTAgnd parasites restores hexosamine-dependent growth. Targeted expression of glucosamine-6-phosphate deaminase, GND, in glycosomes via a canonical C-terminal targeting signal when expressed as a GFP fusion protein

Leishmania major

-

720877

gene GNPDA2, isozyme cGNPDA2-a, DNA and amino acid sequence determination and analysis, four chicken GNPDA2 transcripts, i.e. cGNPDA2-a, cGNPDA2-b, cGNPDA2-c, and cGNPDA2-d, are identified, genetic structure, sequence comparisons and phylogenetic analysis and tree, recombinant overexpression in chicken preadipocytes

gene GNPDA2, isozyme cGNPDA2-b, DNA and amino acid sequence determination and analysis, four chicken GNPDA2 transcripts, i.e. cGNPDA2-a, cGNPDA2-b, cGNPDA2-c, and cGNPDA2-d, are identified, genetic structure, sequence comparisons and phylogenetic analysis and tree, recombinant overexpression in chicken preadipocytes

gene GNPDA2, isozyme cGNPDA2-c, DNA and amino acid sequence determination and analysis, four chicken GNPDA2 transcripts, i.e. cGNPDA2-a, cGNPDA2-b, cGNPDA2-c, and cGNPDA2-d, are identified, genetic structure, sequence comparisons and phylogenetic analysis and tree, recombinant overexpression in chicken preadipocytes

gene GNPDA2, isozyme cGNPDA2-d, DNA and amino acid sequence determination and analysis, four chicken GNPDA2 transcripts, i.e. cGNPDA2-a, cGNPDA2-b, cGNPDA2-c, and cGNPDA2-d, are identified, genetic structure, sequence comparisons and phylogenetic analysis and tree, recombinant overexpression in chicken preadipocytes

gene nagB

gene nagB, coexpression with gene nagA in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain strain XL-1 Blue

gene nagB, recombinant expression of the nonallosteric Bacillus subtilis homologue NagBBs in the NagBEC deficient Escherichia coli mutant, resulting in LAA195 (nagBBs+)

O35000

757097

gene smu.636, cloning from genomic DNA, expression as His-tagged enzyme in Escherichia coli strain BL21(DE3)

Streptococcus mutans

-

two enzyme genes GNP1 and GNP2 in the completely sequenced genomes

the level of cGNPDA2 mRNA is regulated by body weight, feeding/fasting and by consumption of the high-glucose-fat diet, overview