Information on EC 1.5.3.25 - fructosyl amine oxidase (glucosone-forming)

764317

Substrates: -
Products: -

?

deoxy-D-fructosyl-AcGly-Lys + O2 + H2O

? + L-lysine + H2O2

-

Substrates: -
Products: -

?

deoxy-D-fructosyl-AcPhe-Lys + O2 + H2O

? + L-lysine + H2O2

-

Substrates: -
Products: -

?

deoxy-D-fructosyl-Lys-Phe + O2 + H2O

? + L-lysine + H2O2

-

Substrates: -
Products: -

?

deoxy-D-fructosyl-Lys-Ser + O2 + H2O

? + L-lysine + H2O2

-

Substrates: -
Products: -

?

epsilon-fructosyl-aminocaproate + O2 + H2O

glucosone + aminocaproate + H2O2

fructosyl adamantanamine + O2 + H2O

glucosone + adamantanamine + H2O2

fructosyl glycine + O2 + H2O

glucosone + glycine + H2O2

6 entries

fructosyl L-alanine + O2 + H2O

glucosone + L-alanine + H2O2

fructosyl L-leucine + O2 + H2O

glucosone + L-leucine + H2O2

765303

Substrates: -
Products: -

?

fructosyl L-lysine + O2 + H2O

glucosone + L-lysine + H2O2

fructosyl L-phenylalanine + O2 + H2O

glucosone + L-phenylalanine + H2O2

fructosyl L-valine + O2 + H2O

glucosone + L-valine + H2O2

10 entries

fructosyl poly-L-lysine + O2 + H2O

glucosone + poly-L-lysine + H2O2

fructosyl propylamine + O2 + H2O

glucosone + propylamine + H2O2

fructosyl valine + O2 + H2O

glucosone + valine + H2O2

764317

Substrates: -
Products: -

?

fructosyl-L-lysine + O2 + H2O

glucosone + L-lysine + H2O2

Substrates: -
Products: -

?

gamma-fructosyl-aminobutanoate + O2 + H2O

glucosone + aminobutanoate + H2O2

glucose 6-phosphate + alpha-Boc-L-lysine + ?

fructosyl-6-phosphate alpha-Boc-L-lysine + ?

Substrates: -
Products: -

r

glucose 6-phosphate + glycine + ?

fructosyl-6-phosphate glycine + ?

Substrates: -
Products: -

r

glucose 6-phosphate + L-histidine + ?

fructosyl-6-phosphate L-histidine + ?

Substrates: -
Products: -

r

glucose 6-phosphate + L-lysine + ?

fructosyl-6-phosphate L-lysine + ?

Substrates: -
Products: -

r

glucose 6-phosphate + poly-L-lysine + ?

fructosyl-6-phosphate poly-L-lysine + ?

Substrates: -
Products: -

r

Nalpha-fructosyl L-lysine + O2 + H2O

glucosone + L-lysine + H2O2

Aspergillus terreus

Q92204

764320

Substrates: -
Products: -

?

Nalpha-fructosyl-Nepsilon-acetyl-L-lysine + O2 + H2O

glucosone + Nepsilon-acetyl-L-lysine + H2O2

-

Substrates: substrate of isoform amadoriase II
Products: -

?

Nepsilon-(1-deoxy-D-fructos-1-yl)-L-lysine + O2 + H2O

? + L-lysine + H2O2

-

Substrates: -
Products: -

?

Nepsilon-fructosyl L-Lys-L-Leu + O2 + H2O

glucosone + L-Lys-L-Leu + H2O2

-

Substrates: -
Products: -

?

Nepsilon-fructosyl L-Lys-L-Phe + O2 + H2O

glucosone + L-Lys-L-Phe + H2O2

-

Substrates: -
Products: -

?

Nepsilon-fructosyl L-lysine + O2 + H2O

glucosone + L-lysine + H2O2

Nepsilon-fructosyl Nalpha-acetyl-L-Lys-D-Ala-L-Ala + O2 + H2O

glucosone + Nalpha-acetyl-L-Lys-D-Ala-L-Ala + H2O2

-

Substrates: -
Products: -

?

Nepsilon-fructosyl Nalpha-acetyl-L-Lys-D-D-Lys-Sar-Glu + O2 + H2O

glucosone + Nalpha-acetyl-L-Lys-D-Lys-Sar-Glu + H2O2

-

Substrates: -
Products: -

?

Nepsilon-fructosyl Nalpha-acetyl-L-lysine-N-methylamide + O2 + H2O

glucosone + acetyl-L-lysine-N-methylamide + H2O2

-

Substrates: -
Products: -

?

Nepsilon-fructosyl poly-L-lysine + O2 + H2O

glucosone + poly-L-lysine + H2O2

Aspergillus terreus

Q92204

764320

Substrates: poor substrate
Products: -

?

Nepsilon-fructosyl-L-lysine + O2 + H2O

glucosone + L-lysine + H2O2

Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine + O2 + H2O

glucosone + N-alpha-benzyloxycarbonyl-L-lysine + H2O2

Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine + O2 + H2O

glucosone + N-alpha-benzyloxycarbonyl-L-valine + H2O2

-

Substrates: -
Products: -

?

Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-valine + O2 + H2O

glucosone + N-alpha-benzyloxycarbonyl-L-valine + H2O2

-

Substrates: -
Products: -

?

Nepsilon-fructosyl-Nalpha-t-butoxycarbonyl-L-lysine + O2 + H2O

glucosone + Nalpha-t-butoxycarbonyl-L-lysine + H2O2

-

Substrates: -
Products: -

?

S: Nepsilon-fructosyl Nalpha-benzyloxycarbonyl-L-lysine + O2 + H2O

glucosone + Nalpha-benzyloxycarbonyl-L-lysine + H2O2

Aspergillus oryzae

Q60G78, Q60G79

763944

Substrates: -
Products: -

?

additional information

?

-

FAD

6 entries

Ca2+

-

slight activation

Mg2+

-

slight activation

Mn2+

-

slight activation

4-hydroxyphenylglyoxal

-

inhibition of amadoriase II enzyme activity toward fructosyl glycine, with less effect on the enzymatic activity toward fructosyl propylamine

764130

Co2+

-

20 mM, 14% residual activity

fructosyl-thioacetate

-

Ni2+

-

20 mM, 26% residual activity

Zn2+

-

20 mM, 9% residual activity

additional information

-

no inhibiting: EDTA

-

1.7

deoxy-D-fructosyl-AcGly-Lys

-

pH 7.4, 37°C

-

1.5

deoxy-D-fructosyl-AcPhe-Lys

-

pH 7.4, 37°C

-

8.6

deoxy-D-fructosyl-Lys-Phe

-

pH 7.4, 37°C

-

10.3

deoxy-D-fructosyl-Lys-Ser

-

pH 7.4, 37°C

-

6.55 - 27.8

epsilon-fructosyl-aminocaproate

-

3.4 - 14.4

fructosyl adamantanamine

-

0.18 - 9.75

fructosyl glycine

20 entries

-

0.98 - 29.4

fructosyl L-alanine

-

0.71

fructosyl L-phenylalanine

pH 8.0, 37°C

-

0.76 - 29.9

fructosyl L-valine

29 entries

-

0.011 - 3.04

fructosyl propylamine

-

0.34 - 0.99

fructosyl-L-lysine

4 entries

-

9.63 - 47

gamma-fructosyl-aminobutanoate

-

0.22 - 0.51

Nalpha-benzyloxycarbonyl-L-lysine

4.2

Nepsilon-(1-deoxy-D-fructos-1-yl)-L-lysine

-

pH 7.4, 37°C

-

9.8 - 63

Nepsilon-fructosyl-L-lysine

-

0.15 - 29

Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine

-

1.6 - 3

Nepsilon-fructosyl-Nalpha-t-butoxycarbonyl-L-lysine

-

1.4

deoxy-D-fructosyl-AcGly-Lys

-

pH 7.4, 37°C

-

1.8

deoxy-D-fructosyl-AcPhe-Lys

-

pH 7.4, 37°C

-

0.5

deoxy-D-fructosyl-Lys-Phe

-

pH 7.4, 37°C

-

0.3

deoxy-D-fructosyl-Lys-Ser

-

pH 7.4, 37°C

-

2.91 - 10.6

epsilon-fructosyl-aminocaproate

-

0.007 - 11.9

fructosyl adamantanamine

-

0.6 - 8

fructosyl glycine

9 entries

-

0.7 - 21

fructosyl L-alanine

-

0.04 - 69.8

fructosyl L-valine

14 entries

-

0.008

fructosyl poly-L-lysine

wild-type, pH 8, temperature not specified in the publication

-

0.08 - 5.8

fructosyl propylamine

-

11.97 - 22.9

fructosyl-L-lysine

4 entries

-

1.55 - 9.73

gamma-fructosyl-aminobutanoate

-

1.1

Nepsilon-(1-deoxy-D-fructos-1-yl)-L-lysine

-

pH 7.4, 37°C

-

2.48 - 27.5

Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine

-

4.2 - 5.5

Nepsilon-fructosyl-Nalpha-t-butoxycarbonyl-L-lysine

-

0.11 - 1.62

epsilon-fructosyl-aminocaproate

-

0.06 - 0.67

fructosyl adamantanamine

-

0.066 - 15.6

fructosyl glycine

9 entries

-

0.24 - 21.4

fructosyl L-alanine

-

4.2 - 10.8

fructosyl L-lysine

-

0.002 - 68.9

fructosyl L-valine

14 entries

-

0.28 - 325

fructosyl propylamine

-

12 - 64.2

fructosyl-L-lysine

4 entries

-

0.03 - 1.01

gamma-fructosyl-aminobutanoate

-

6.36

Nepsilon-fructosyl L-Lys-L-Leu

-

pH 7.4, 37°C

-

5.6

Nepsilon-fructosyl L-Lys-L-Phe

-

pH 7.4, 37°C

-

9.6

Nepsilon-fructosyl Nalpha-acetyl-L-Lys-D-Ala-L-Ala

-

pH 7.4, 37°C

-

3.54

Nepsilon-fructosyl Nalpha-acetyl-L-lysine-N-methylamide

-

pH 7.4, 37°C

-

0.19 - 106

Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine

-

0.33 - 1.33

Nepsilon-fructosyl-Nalpha-t-butoxycarbonyl-L-lysine

-

0.09

mutant N47A, pH 7, 23°C

764361

0.52

wild-type, pH 7, 23°C

6.8

isoform FAOD-Ao1

7.5

-

wild-type

7.8

-

8

8.2

isoform FAOD-Ao2

8.3

-

8.5

-

amadoriase II

35

-

37

-

wild-type

40

50

mutant T60A/A188G/M244L/N257S/L261M

763943

40

less than 20% residual activity

55

wild-type is inactive

4.6

isoelectric focusing

5.5 - 5.7

-

amadoriase I, isoelectric focusing

6.7

-

amadoriase II, isoelectric focusing

-

UniProt

-

UniProt

-

764126, 764128, 764130, 765033

-

-

UniProt

-

UniProt

-

-

-

-

-

764360, 764361, 765303, 765700

UniProt

-

-

-

-

Highest Expressing Human Cell Lines

Filter by:

Cell Line Links

Gene Links

metabolism

physiological function

39000

gel filtration

40000

-

gel filtration, isoform amadoriase I

55000

-

gel filtration, isoform amadoriase II

85000

gel filtration

88000

gel filtration

98000

gel filtration

?

dimer

monomer

mutant T60A/A188G/M244L/N257S/L261M, giving rhombic prismatic yellow crystals, space group C2 with unit-cell parameters a 101.08, b 63.36, c 83.07 A, beta 108.8°, to at least 1.8 A resolution, and additionally rod-like crystals, space group P4122 or P4322 with unitcell parameters a = b = 119.09, c 164.66 A and diffract to 2.7 A resolution

763847

structure of FAOX-II shows an interaction between Lys53 and the isoalloxazine. The ammonium nitrogen of the lysine is in contact with and nearly centered over the aromatic ring of the flavin on the si-face. The positive charge of Lys53 is critical for flavin reduction, but plays very little role in the reaction with molecular oxygen

764132

structures in free form and in complex with the inhibitor fructosyl-thioacetate, at 1.75 and 1.6 A resolution, respectively. FAOX-II is a two domain FAD-enzyme with an overall topology similar to that of monomeric sarcosine oxidase. Active site residues Tyr-60, Arg-112 and Lys-368 bind the carboxylic portion of the fructosamine, whereas Glu-280 and Arg-411 bind the fructosyl portion

structures of the free and the substrate-bound form of amadoriase I and comparison with isoform amadoriase II. While in Amadoriase II the loop 59-70 (amadoriase I numbering) is directed toward the core of the protein, in amadoriase I the loop is projected outward and is exposed to the solvent, contributing to the formation of a larger catalytic cavity

765733

homology modeling and docking of substrates fructosyl L-valine and Nepsilon-fructosyl-L-lysine. Residue Asn354 interacts closely with Nepsilon-fructosyl-L-lysine, but not with fructosyl L-valine

homology modeling of structure. Asn354 plays an important role in substrate recognition and can be substituted in order to change substrate specificity while maintaining high catalytic activity

T60A/A188G/M244L/N257S/L261M

mutant with improved thermostability, stable at 45°C

763943

D295C/K303C

mutation leads to increase in melting temperature, mutant shows hyperstabilization with residual activity up to 95°C

E280L

mutation in fructosyl binding site, loss of activity

E71R

activity similar to wild-type

E71R/R114E

8fold increase in Km value

E71S

mutation in salt bridge between flexible regions, activity similar to wild-type

E75N/R108L7R114W/W208R/Q289R/T303S

8.78fold increase in the activity toward fructosyl-polylysine and also performs several fold better than the wild-type on real gravy stains at concentrations of 10-100 microg/ml

764003

H106S/G150S

mutation leads to decrease in melting temperature

H357N

inactive

K368M

5fold increase in Km vlaue

K53M

more than a million-fold decrease in flavin reduction, while only slowing the oxygen reaction by about 30fold

764132

K53R

mutation has minor effects on catalysis

764132

N52A

mutation results in significant loss in the catalytic ability to employ O2 as the electron acceptor, while having little effect on the dye-mediated dehydrogenase activity employing artificial electron acceptors instead of O2

764361

N63D/K64Q/R112W/V113F/R114S

strong increase in activity with fructosyl adamantanamine and with fructosyl polylysine

R112E

10fold increase in Km value

R112W

strong increase in activity with fructosyl adamantanamine

R112Y

strong increase in activity with fructosyl adamantanamine

R112Y/V113F/R114S

strong increase in activity with fructosyl adamantanamine

R114E

10fold increase in Km value

R114S

slight decrease in Km value

R411L

loss of activity

S370A

inactive

S67C/P121C

mutation leads to increase in melting temperature

Y60F

mutation in carboxylate binding site, increase in Km value

C342A

-

about 2% of wild-type activity, loss of covalent binding of FAD

764126

C342S

-

less than 1% of wild-type activity, loss of covalnet binding of FAD

764126

R112E

R112E/R114E

-

mutation in isoform II, kcat value is similar to the wild-type, while the Km of the mutant toward fructosyl glycine increases significantly

764130

R114E

K373M

-

catalytic turnover for fructosyl valine greatly decreases, whereas that for Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine does not

K373R

-

mutant has lost the activity on substrate fructosyl valine

K373T

-

catalytic turnover for fructosyl valine greatly decreases, whereas that for Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine does not

K373V

-

catalytic turnover for fructosyl valine greatly decreases, whereas that for Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine does not

K373W

-

catalytic turnover for fructosyl valine greatly decreases, whereas that for Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine does not

K373M

-

catalytic turnover for fructosyl valine greatly decreases, whereas that for Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine does not

-

K373R

-

mutant has lost the activity on substrate fructosyl valine

-

K373T

-

catalytic turnover for fructosyl valine greatly decreases, whereas that for Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine does not

-

K373V

-

catalytic turnover for fructosyl valine greatly decreases, whereas that for Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine does not

-

K373W

-

catalytic turnover for fructosyl valine greatly decreases, whereas that for Nepsilon-fructosyl-Nalpha-benzyloxycarbonyl-L-lysine does not

-

C324A

mutation of putative active site residue, no residual activity with fructosyl L-valine

F356A

mutation of putative active site residue, 18% of wild-type activity with fructosyl L-valine

K357A

mutation of putative active site residue, 2% of wild-type activity with fructosyl L-valine

K48A

mutation of putative active site residue, no residual activity with fructosyl L-valine

N354A

N354H

mutation leads to an increase in the activity ratio fructosyl L-valine/Nepsilon-fructosyl-L-lysine

N354K

mutation leads to an increase in the activity ratio fructosyl L-valine/Nepsilon-fructosyl-L-lysine

N354V

mutation leads to an increase in the activity ratio fructosyl L-valine/Nepsilon-fructosyl-L-lysine

N44A

mutation of putative active site residue, 4% of wild-type activity with fructosyl L-valine

N47A

N47D

loss of activity

R94F

-

mutation enhances substrate specificity toward fructosyl valine

R94L

-

mutation enhances substrate specificity toward fructosyl valine

R94W

-

specificity of mutant enzyme toward fructosyl valine is 14fold that of the wild-type enzyme

40.8

melting temperature, mutant H106S/G150S

45

-

1 h, 24% residual activity for wild-type, 80% for mutants R94W, R94F, R94L

50

half-life of wild-type 2.9 min, of mutant T60A/A188G/M244L/N257S/L261M 45 min, respectively

763943

52.4

melting temperature, wild-type

55.3

melting temperature, mutant S67C/P121C

60.6

melting temperature, mutant D295C/K303C

-

recombinant protein, purified by Ni His-tag affinity chromatography

-

-

expression in Escherichia coli

7 entries

expression of isoform amadoriase I in Escherichai coli

-

expression is inducible by fructosyl amines and by the VeA gene, encoding an activator of sexual development, the veA gene is necessary for full induction of FaoA gene expression by fructosyl amines

Aspergillus nidulans

Q96UT4

764042

expression of isoform AoFAO1 by fructosyl L-valine depends on the expression of AoFAO2. Both genes are also induced by reaction product, glucosone

Aspergillus oryzae

Q60G78, Q60G79

764681

analysis

development of a continuous, coupled enzyme assay

synthesis

-

addition of amadoriase I during peptide glycation reaction reduces the formation of Amadori products up to 80%. The effect is more evident for hydrophobic peptides. In protein-glucose systems, the effect is dependent on the molecular weight and steric hindrance being negligible for BSA and at a maximum for insulin