Information on EC 3.4.21.63 - Oryzin

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.21.63
-
RECOMMENDED NAME
GeneOntology No.
Oryzin
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of proteins with broad specificity, and of Bz-Arg-OEt > Ac-Tyr-OEt. Does not hydrolyse peptide amides
show the reaction diagram
broad specificity
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
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CAS REGISTRY NUMBER
COMMENTARY hide
9074-07-1
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain NCIM637
-
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Manually annotated by BRENDA team
strain NCIM637
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Manually annotated by BRENDA team
gene prtA
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Manually annotated by BRENDA team
gene prtA
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Manually annotated by BRENDA team
strain ATCC16168
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Manually annotated by BRENDA team
strain AWT20
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Manually annotated by BRENDA team
EI212
-
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Manually annotated by BRENDA team
NRRL 2160
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Manually annotated by BRENDA team
strain U152
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Manually annotated by BRENDA team
strain Y2942
SwissProt
Manually annotated by BRENDA team
NTU-163
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Manually annotated by BRENDA team
induced by growth on modified Reeses medium with 0.5% casein, 0.25% glucose and 0.05% yeast extract, pH 8.5
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
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the enzyme belongs to the gluzincin family
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
3-phenyllactic acid + H2O
?
show the reaction diagram
-
methyl esters and its ring-substituted derivative hydrolyzed with excellent enantioselectivities
-
-
?
4-nitrophenyl acetate + H2O
4-nitrophenol + acetate
show the reaction diagram
Acetyl-Ala methyl ester + H2O
Acetyl-Ala + methanol
show the reaction diagram
Acetyl-Ala-Ala methyl ester + H2O
Acetyl-Ala-Ala + methanol
show the reaction diagram
Acetyl-Ala-Ala-Ala methyl ester + H2O
Acetyl-Ala-Ala-Ala + methanol
show the reaction diagram
Acetyl-Ala-Ala-Ala-Ala methyl ester + H2O
Acetyl-Ala-Ala-Ala-Ala + methanol
show the reaction diagram
Acetyl-Ala-Ala-Phe methyl ester + H2O
Acetyl-Ala-Ala-Phe + methanol
show the reaction diagram
Acetyl-Ala-Phe methyl ester + H2O
Acetyl-Ala-Phe + methanol
show the reaction diagram
Acetyl-Phe methyl ester + H2O
Acetyl-Phe + methanol
show the reaction diagram
azocasein + H2O
?
show the reaction diagram
-
-
-
-
?
Benzoylarginine ethyl ester + H2O
?
show the reaction diagram
-
-
-
-
-
Benzyloxycarbonyl-Ala-Ala-Lys methyl ester + H2O
Benzyloxycarbonyl-Ala-Ala-Lys + methanol
show the reaction diagram
Benzyloxycarbonyl-Ala-Lys methyl ester + H2O
Benzyloxycarbonyl-Ala-Lys + methanol
show the reaction diagram
Benzyloxycarbonyl-Gly-Lys methyl ester + H2O
Benzyloxycarbonyl-Gly-Lys + methanol
show the reaction diagram
Benzyloxycarbonyl-Leu-Lys methyl ester + H2O
Benzyloxycarbonyl-Leu-Lys + methanol
show the reaction diagram
Benzyloxycarbonyl-Lys methyl ester + H2O
Benzyloxycarbonyl-Lys + methanol
show the reaction diagram
Benzyloxycarbonyl-Phe-Lys methyl ester + H2O
Benzyloxycarbonyl-Phe-Lys + methanol
show the reaction diagram
Carbobenzoxy-Leu-Gly-NH2 + H2O
Carbobenzoxy-Leu-Gly + NH3
show the reaction diagram
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-
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Carbobenzoxy-Pro-Leu-NH2 + H2O
Carbobenzoxy-Pro-Leu + NH3
show the reaction diagram
-
-
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casein + H2O
?
show the reaction diagram
casein + H2O
hydrolyzed casein
show the reaction diagram
defatted soybean complex medium + H2O
?
show the reaction diagram
Egg albumin + H2O
Hydrolyzed egg albumin
show the reaction diagram
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-
-
-
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Fibrin + H2O
Hydrolyzed fibrin
show the reaction diagram
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-
-
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Fibrinogen + H2O
Hydrolyzed fibrinogen
show the reaction diagram
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-
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Gelatin + H2O
Hydrolyzed gelatin
show the reaction diagram
Hemoglobin + H2O
Hydrolyzed hemoglobin
show the reaction diagram
Ile-Gln-Asn-Cys-Pro-Leu-Gly-NH2 + H2O
Ile-Gln-Asn-Cys-Pro-Leu + Gly-NH2
show the reaction diagram
-
-
-
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L-Leu 4-nitroanilide hydrochloride + H2O
?
show the reaction diagram
-
-
-
-
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mandelic acid + H2O
?
show the reaction diagram
-
only low enantioselectivities
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-
?
N-Acetyl amino acid ester + H2O
?
show the reaction diagram
-
hydrolysis decreases in the order of Phe/ Tyr, Trp, Met, Leu, Lys, His, Val/ Gly
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N-Acetyl-L-tyrosine ethyl ester + H2O
?
show the reaction diagram
N-Benzoyl-L-arginine ethyl ester + H2O
?
show the reaction diagram
-
-
-
-
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N-Cbz-L-Asp + sec-butanol
N-Cbz-L-Asp-sec-butyl ester
show the reaction diagram
-
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r
N-Cbz-L-Tyr + 1-phenylethanol
N-Cbz-L-Tyr-1-phenylethyl ester
show the reaction diagram
-
-
-
?
N-Cbz-L-Tyr + sec-butanol
N-Cbz-L-Tyr-sec-butyl ester
show the reaction diagram
-
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r
Oxidized insulin A-chain + H2O
Hydrolyzed oxidized insulin A-chain
show the reaction diagram
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at 0C the enzyme cleaves only the bond Glu17-Asn18
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Oxidized insulin B-chain + H2O
?
show the reaction diagram
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eight cleavage sites of the enzyme in oxidized insulin B-chain are determined by mass spectrometry, and five of them have high hydrophobic amino acid affinity
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?
Oxidized insulin B-chain + H2O
Hydrolyzed oxidized insulin B-chain
show the reaction diagram
Poly-L,alpha-glutamic acid + H2O
?
show the reaction diagram
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-
-
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Poly-L-lysine + H2O
?
show the reaction diagram
-
-
-
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Protamine + H2O
Hydrolyzed protamine
show the reaction diagram
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-
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Serum albumin + H2O
Hydrolyzed serum albumin
show the reaction diagram
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Soybean 11S globulin + H2O
Hydrolyzed soybean 11S globulin
show the reaction diagram
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-
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Soybean protein + H2O
Hydrolyzed soybean protein
show the reaction diagram
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-
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additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Cu2+
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activation
Fe2+
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activates 1.74fold at 5 mM
K+
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activates about 10% at 5 mM
Na+
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activates about 10% at 5 mM
Zn2+
-
the enzyme has a zinc-binding motif and is a gluzincin
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-nitrophenyl acetate
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increasing concentrations of the ester inhibit casein hydrolysis
Ba2+
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some inhibition at 10 mM
benzamidine
slightly inhibits recombinant and native enzyme
benzeneboronic acid
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-
Benzyloxycarbonyl-Ala-Gly-PheCH2Cl
Ca2+
-
some inhibition at 10 mM
Cd2+
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some inhibition at 1 mM
ethanol
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reversible inhibition
leupeptin
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68% inhibition of AP30 at 0.05 mM
methanol
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inhibition of esterolytic activity
N-bromosuccinimide
n-butanol
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inhibition of esterolytic activity
N-Chlorosuccinimide
-
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n-Propanol
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inhibition of esterolytic activity
N-tosyl-L-lysine chloromethyl ketone
inhibitor of recombinant and native enzyme
NaN3
-
slightly inhibits
Pb2+
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some inhibition at 10 mM
Phenylmethyl sulfonylfluoride
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complete inhibition
phenylmethylsulfonyl fluoride
Potato inhibitor
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sodium lauryl sulfate
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Sr2+
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some inhibition at 10 mM
TLCK
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moderately inhibits
TPCK
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moderately inhibits
Trypsin inhibitor
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moderately inhibits
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additional information
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
sulfhydryl compounds
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accelerate the action
additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
44.5 - 68.5
acetyl-(Ala)2 methyl ester
6.7 - 32.1
acetyl-(Ala)3 methyl ester
4.7 - 194
acetyl-(Ala)4 methyl ester
96.8
Acetyl-Ala methyl ester
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Aspergillus sojae
13.5
Acetyl-Ala-Phe methyl ester
-
Aspergillus sojae
23
Acetyl-Phe methyl ester
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Aspergillus sojae
12.8
acetyl-tyrosine ethyl ester
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Aspergillus oryzae
7.1
benzoyl-arginine ethyl ester
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Aspergillus oryzae
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1150 - 2000
acetyl-(Ala)2 methyl ester
1540 - 3720
acetyl-(Ala)3 methyl ester
1970 - 2090
acetyl-(Ala)4 methyl ester
28.3 - 28.9
Acetyl-Ala methyl ester
43
acetyltyrosine ethyl ester
Aspergillus oryzae
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Aspergillus oryzae
41
benzoylarginine ethyl ester
Aspergillus oryzae
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Aspergillus oryzae
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25.7
-
crude extract
40.94
culture supernatant
92.73
purified recombinant enzyme
750
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30fold purified enzyme
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
-
assay at
7.5 - 8
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N-carbobenzoxyglycine 4-nitrophenyl ester
7.5
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carbobenzoxy-Gly-Leu-NH2
7.8
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gelatin
8 - 8.5
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casein
8.5 - 8.8
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N-benzoyl-L-arginine ethyl ester
9
recombinant and native enzyme
9 - 10
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Aspergillus sojae
10
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casein, hemoglobin
10.5
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milk casein
11 - 11.5
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milk casein
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10
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reduction in enzyme activity is 19% at pH 7.0 and 36% at pH 10.0
6 - 11
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small changes of activity between pH 6 and 11
7 - 10
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
43
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at pH 8.0, milk casein
47
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at pH 7.0, milk casein
50 - 55
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casein
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 55
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100% activity at 40C while 92% residual activity at 45C, 74% at 50C, and 41% 55C, inactive above 55C, temperature profile, overview
35 - 60
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activity is reduced by 15% at 50C and by 66% at 55C
40 - 60
recombinant and native enzyme
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
15500
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x * 15500, AP15, SDS-PAGE and mass spectrometry, x * 30000, AP30, SDS-PAGE and mass spectrometry
18000
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Aspergillus flavus
22000 - 24000
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Aspergillus flavus, gel filtration
22000
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Aspergillus candidus, calculation from amino acid composition
23620 - 27180
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Aspergillus flavus, ultracentrifugal measurements
25000
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Aspergillus oryzae, gel filtration
25500
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Aspergillus sojae
30000
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x * 15500, AP15, SDS-PAGE and mass spectrometry, x * 30000, AP30, SDS-PAGE and mass spectrometry
33000
-
gel filtration
34000
x * 34000, recombinant enzyme, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 11
-
4C, stable
29487
5 - 8.5
5 - 9
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purified recombinant truncated neutral protease I, stable at
731961
5.5 - 10
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immobilized enzyme, shows enhanced stability in acidic as well as alkaline environments in comparison to the free enzyme (stable between pH 7.0-9.0)
680260
6
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50C, stable
29487
6 - 12
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671008
7 - 12
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partially purified enzyme, stable at
717055
7.5
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35C, highest stability
29484
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 40
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partially purified enzyme, stable at, retention of 96% activity after 1 h of incubation at 40C
35
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pH 7.5, highest stability
40
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half-life 90 min
50
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half-life 18 min
50 - 55
-
activity is reduced by 15% at 50C and by 66% at 55C
50
-
purified recombinant truncated neutral protease I, 120 min, over 90% activity remaining
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
2 mM Ca2+ protect against heat denaturation
-
Enzyme is unstable in 8 M urea, overnight at 30C
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acetone
benzene
-
89.2% remaining activity after 1 h at 30C in 25% organic solvent
Butanol
chloroform
-
105.3% activity after 1 h at 30C in 25% organic solvent
diethyl ether
-
89.2% remaining activity after 1 h at 30C in 25% organic solvent
Ethanol
hexane
-
94.3% remaining activity after 1 h at 30C in 25% organic solvent
Methanol
toluene
-
91.7% remaining activity after 1 h at 30C in 25% organic solvent
xylene
-
83.6% remaining activity after 1 h at 30C in 25% organic solvent
additional information
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
oxidant stable serine protease. The enzyme retains 100% of its original activity with 10% H2O2 after 1 h of incubation at 30C
-
717055
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
AP30 purified by cationic exchange resin followed by an affinity column, AP15 partially purified using an anion-exchange column
-
by ammonium sulfate precipitation, ion exchange chromatography and gel filtration
native enzyme partially by acetone precipitation
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partially purified by ion exchange chromatography and gel filtration, 30fold
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var. columnaris, partial
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gene prtA, DNA and amino acid sequence determination and analysis, expression in Pichia pastoris strain X-33 as inactive enzyme, the presence of this enzyme was toxic for the host and resulted in cell lysis, modification of the culture medium constituents improve the expression by 6.4fold
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high-level expression of plasmids pPIC9K-n/Alp and pPIC9K-alpha/Alp in Pichia pastoris GS115 with native signal peptide or alpha-factor secretion signal peptide
overexpression in Escherichia coli strain BL21(DE3)
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recombinant expression of a truncated neutral protease I in Pichia pastoris with a high enzyme yield
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sequence comparisons between Aspergillus species and different proteases, clustering of alkaline proteases, phylogenetic tree, overview
sequence comparisons between Aspergillus species and different proteases, clustering of alkaline proteases, phylogenetic tree, overview; sequence comparisons between Aspergillus species and different proteases, clustering of alkaline proteases, phylogenetic tree, overview
strain U1521 contains multiple copies of alkaline protease gene
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the enzyme is not induced by lipids
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
detergent
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enzyme is highly compatible with commercial detergents such as Vim and Avis and may find application in laundry detergents
food industry
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the enzyme is efficient in producing antihypertensive peptide IPP from beta-casein and a potential debittering agent. The high degree of hydrolysis of the enzyme to soybean protein (8.8%) and peanut protein (11.1%) compared to papain and alcalase makes it a good candidate in the processing of oil industry byproducts
industry
nutrition