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Information on EC 3.4.23.4 - chymosin and Organism(s) Bos taurus and UniProt Accession P00794

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EC Tree
     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.23 Aspartic endopeptidases
                3.4.23.4 chymosin
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This record set is specific for:
Bos taurus
UNIPROT: P00794 not found.
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The taxonomic range for the selected organisms is: Bos taurus
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
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Broad specificity similar to that of pepsin A. Clots milk by cleavage of a single Ser-Phe105-/-Met-Ala bond in kappa-chain of casein
Synonyms
chymosin, prochymosin, rennin, chymosin a, chymosin b, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
CHY-MAX
-
commercial preparation
CHY-MAX 200
-
commercial preparation
chymase
-
-
-
-
chymosin A
-
allelic form
chymosin B
Preprorennin
-
-
-
-
prochymosin
rCH
-
recombinant enzyme
rennin
CAS REGISTRY NUMBER
COMMENTARY hide
9001-98-3
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
AFPLEFEREL + H2O
AFPLEF + EREL
show the reaction diagram
modified peptide substrate based on residues 165-174 of proopiomelanocortin
-
-
?
AFPLEFFREL + H2O
AFPLEF + FREL
show the reaction diagram
modified peptide substrate based on residues 165-174 of proopiomelanocortin
-
-
?
AFPLEFIREL + H2O
AFPLEF + IREL
show the reaction diagram
modified peptide substrate based on residues 165-174 of proopiomelanocortin
-
-
?
AFPLEFKREL + H2O
AFPLEF + KREL
show the reaction diagram
modified peptide substrate based on residues 165-174 of proopiomelanocortin
-
-
?
kappa-casein + H2O
?
show the reaction diagram
Abz-A-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine + H2O
?
show the reaction diagram
-
low molecular weight, fluorogenic peptide substrate
-
-
?
Abz-A-A-F-F-A-A-p-nitroanilide + H2O
?
show the reaction diagram
-
low molecular weight, fluorogenic peptide substrate
-
-
?
Abz-A-A-F-F-A-N-(2,4-dinitrophenyl)-ethylenediamine + H2O
?
show the reaction diagram
-
low molecular weight, fluorogenic peptide substrate
-
-
?
Abz-A-A-F-F-N-(2,4-dinitrophenyl)-ethylenediamine + H2O
?
show the reaction diagram
-
low molecular weight, fluorogenic peptide substrate
-
-
?
Abz-A-A-F-F-pnA + H2O
?
show the reaction diagram
-
low molecular weight, fluorogenic peptide substrate
-
-
?
Abz-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine + H2O
?
show the reaction diagram
-
low molecular weight, fluorogenic peptide substrate
-
-
?
acid denatured hemoglobin + H2O
?
show the reaction diagram
-
-
-
-
?
alpha-casein + H2O
?
show the reaction diagram
alphaS-casein + H2O
?
show the reaction diagram
-
-
-
-
?
alphas1-casein + H2O
?
show the reaction diagram
-
-
-
-
?
azocasein + H2O
?
show the reaction diagram
-
-
-
-
?
beta-casein + H2O
?
show the reaction diagram
beta-chain of proteolytic insulin + H2O
?
show the reaction diagram
-
general proteolytic activity
-
-
?
bovine kappa-casein + H2O
?
show the reaction diagram
bovine kappa-casein + H2O
para-kappa-casein + caseinomacropeptide
show the reaction diagram
-
the enzyme cleaves the Phe105-Met106 bond of kappa-casein
-
-
?
bovine kappa-casein residues 97-112 + H2O
?
show the reaction diagram
-
substrate binds in an extended conformation with charged residues on either side of the scissile bond playing an important role in stabilizing the binding pose. Substrate residues Lys111 and Lys112 bind to the N-terminal domain of chymosin displacing a conserved water molecule. A cluster of histidine and proline residues, His98-Pro99-His100-Pro101-His102, in kappa-casein binds to the C-terminal domain of the protein, where neighboring conserved arginine residue Arg97 is important for stabilizing the binding pose. The catalytic site including the catalytic water molecule is stable in the starting conformation of the general acid/base catalytic mechanism for 18 ns of molecular dynamics simulations
-
-
?
casein + H2O
?
show the reaction diagram
His-Pro-His-Pro-His-Leu-Ser-Phe-Met-Ala-Ile-Pro-NH2 + H2O
His-Pro-His-Pro-His-Leu-Ser-Phe + Met-Ala-Ile-Pro + NH3
show the reaction diagram
His-Pro-His-Pro-His-Leu-Ser-Phe-Phe(NO2)-Ala-Ile-Pro-Pro-Lys-Lys + H2O
?
show the reaction diagram
-
-
-
-
?
HPHPHLSFMAIPPKK + H2O
?
show the reaction diagram
-
-
-
-
?
kappa-casein + H2O
?
show the reaction diagram
kappa-casein + H2O
casein macropeptide + para-kappa-casein
show the reaction diagram
kappa-casein + H2O
caseinmacropeptide + para-kappa-casein
show the reaction diagram
kappa-casein + H2O
p-kappa-casein + glycomacropeptide
show the reaction diagram
kappa-casein + H2O
para-kappa-casein + macropeptide
show the reaction diagram
-
kappa-casein is the primary substrate for the chymosin action
-
-
?
L-leucine-4-nitroanilide + H2O
L-leucine + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
L-S-F-M-A-I-P-NH2 + H2O
?
show the reaction diagram
-
hepapeptide, a fragment of the native chymosin substrate kappa-casein, is most efficiently cleaved by native calf chymosin and less efficiently by transgenic chymosin and recombinant chymosin
-
-
?
Leu-Ser-Phe(NO2)-Nle-Ala-Leu-OMe + H2O
?
show the reaction diagram
-
-
-
-
?
Leu-Ser-Phe-Met-Ala-Ile-O-methyl ester + H2O
Leu-Ser-Phe + Met-Ala-Ile-O-methyl ester
show the reaction diagram
-
part of the bovine kappa-casein sequence
-
?
Leu-Ser-Phe-Met-Ala-Ile-Pro-NH2 + H2O
Leu-Ser-Phe + Met-Ala-Ile-Pro + NH3
show the reaction diagram
Leu-Ser-Phe-Met-Ala-O-methyl ester + H2O
Leu-Ser-Phe + Met-Ala-O-methyl ester
show the reaction diagram
-
part of the bovine kappa-casein sequence
-
?
Lys-Pro-Ala-Glu-Phe-Phe(NO2)-Ala-Leu-OH + H2O
Lys-Pro-Ala-Glu-Phe + Phe(NO2)-Ala-Leu
show the reaction diagram
Lys-Pro-Leu-Glu-Phe-Phe(NO2)-Arg-Leu + H2O
Lys-Pro-Leu-Glu-Phe + Phe(NO2)-Arg-Leu
show the reaction diagram
-
-
-
?
milk + H2O
?
show the reaction diagram
o-aminobenzoyl-Ala-Ala-Phe-Phe-Ala-Ala-NH-C6H4NO2 + H2O
o-aminobenzoyl-Ala-Ala-Phe + Phe-Ala-Ala-NHC6H4NO2
show the reaction diagram
-
-
-
?
o-aminobenzoyl-Ala-Ala-Phe-Phe-NH-C6H4-NO2 + H2O
o-aminobenzoyl-Ala-Ala-Phe + Phe-NH-C6H4-NO2
show the reaction diagram
-
-
-
?
Pro-His-Leu-Ser-Phe-Met-Ala-Ile-O-methyl ester + H2O
Pro-His-Leu-Ser-Phe + Met-Ala-Ile-O-methyl ester
show the reaction diagram
-
part of the bovine kappa-casein sequence
-
?
Ser-Phe-Met-Ala-Ile-O-methyl ester + H2O
Ser-Phe + Met-Ala-Ile-O-methyl ester
show the reaction diagram
-
part of the bovine kappa-casein sequence
-
?
skim milk + H2O
?
show the reaction diagram
-
-
-
-
?
skimmed milk + H2O
?
show the reaction diagram
-
-
-
-
?
undecapeptide analogue to chymosin sensitive region of bovine kappa casein
?
show the reaction diagram
-
synthetic substrate
-
-
?
undecapeptide analogue to chymosin sensitive region of camel kappa casein
?
show the reaction diagram
-
synthetic substrate
-
-
?
whole milk + H2O
?
show the reaction diagram
-
-
-
-
?
YGISSKFCE + H2O
YGISSKF + L-Cys-L-Glu
show the reaction diagram
-
modified peptide based on prochymosin sequence
100% cleavage at enzyme to substrate ratio of 1:1, pH 6.2, 16 h
-
?
YGISSKFfE + H2O
YGISSKF + FE
show the reaction diagram
-
modified peptide based on prochymosin sequence
100% cleavage at enzyme to substrate ratio of 1:1, pH 6.2, 16 h
-
?
YGISSKFHE + H2O
YGISSKF + His-Glu
show the reaction diagram
-
modified peptide based on prochymosin sequence
51% cleavage at enzyme to substrate ratio of 1:1, pH 6.2, 16 h
-
?
YGISSKFIE + H2O
YGISSKF + Ile-Glu
show the reaction diagram
-
modified peptide based on prochymosin sequence
54% cleavage at enzyme to substrate ratio of 1:1, pH 6.2, 16 h
-
?
YGISSKFKE + H2O
YGISSKF + Lys-Glu
show the reaction diagram
-
modified peptide based on prochymosin sequence
33% cleavage at enzyme to substrate ratio of 1:1, pH 6.2, 16 h
-
?
YGISSKFLE + H2O
YGISSKF + L-Leu-L-Glu
show the reaction diagram
-
modified peptide based on prochymosin sequence
42% cleavage at enzyme to substrate ratio of 1:1, pH 6.2, 16 h
-
?
YGISSKFME + H2O
YGISSKF + L-Met-L-Glu
show the reaction diagram
-
modified peptide based on prochymosin sequence
52% cleavage at enzyme to substrate ratio of 1:1, pH 6.2, 16 h
-
?
YGISSKFNE + H2O
YGISSKF + L-Asn-L-Glu
show the reaction diagram
-
modified peptide based on prochymosin sequence
33% cleavage at enzyme to substrate ratio of 1:1, pH 6.2, 16 h
-
?
YGISSKFRE + H2O
YGISSKF + Arg-Glu
show the reaction diagram
-
modified peptide based on prochymosin sequence
59% cleavage at enzyme to substrate ratio of 1:1, pH 6.2, 16 h
-
?
YGISSKFVE + H2O
YGISSKF + Val-Glu
show the reaction diagram
-
modified peptide based on prochymosin sequence
36% cleavage at enzyme to substrate ratio of 1:1, pH 6.2, 16 h
-
?
YGISSKFWE + H2O
YGISSKF + Trp-Glu
show the reaction diagram
-
modified peptide based on prochymosin sequence
100% cleavage at enzyme to substrate ratio of 1:1, pH 6.2, 16 h
-
?
YGISSKFYE + H2O
YGISSKF + L-Tyr-L-Glu
show the reaction diagram
-
modified peptide based on prochymosin sequence
100% cleavage at enzyme to substrate ratio of 1:1, pH 6.2, 16 h
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
kappa-casein + H2O
?
show the reaction diagram
alphas1-casein + H2O
?
show the reaction diagram
-
-
-
-
?
bovine kappa-casein + H2O
para-kappa-casein + caseinomacropeptide
show the reaction diagram
-
the enzyme cleaves the Phe105-Met106 bond of kappa-casein
-
-
?
casein + H2O
?
show the reaction diagram
-
the enzyme breaks the bond between Phe-105 and Met-106 in the casein and initiates the coagulation of milk
-
-
?
kappa-casein + H2O
?
show the reaction diagram
kappa-casein + H2O
casein macropeptide + para-kappa-casein
show the reaction diagram
-
cleaves a single bond between phenylalanine 105 and methionine 106
-
-
?
kappa-casein + H2O
caseinmacropeptide + para-kappa-casein
show the reaction diagram
-
cleaves a single bond between phenylalanine 105 and methionine 106
-
-
?
milk + H2O
?
show the reaction diagram
-
the maximum increase on milk-clotting activity is 10% for recombinant chymosin at 212MPa/5 min/10°C
-
-
?
skim milk + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
the enzyme has milk coagulation bioactivity
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-2-epoxy-3(p-nitrophenoxy)propane
-
-
Blood serum
-
porcine and particularly equine serum impairs the coagulation of milk by chymosin, the inhibitor(s) may be heat labile, alpha2-macroglobulin may be the inhibitory substance
-
Ca2+
-
Ca2+ inhibits milk-clotting activity (about 35% residual activity at 5 mM)
Diazoacetylnorleucine methyl ester
-
in presence of Cu2+
pepstatin
pepstatin A
-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
kappa-casein
a HPHPH sequence in the P8-P4 residues of the natural substrate kappa-casein acts as the allosteric activator of the enzyme
-
His-Pro-His-Pro-His-NH2
-
stimulation, part of kappa-casein
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.3
AFPLEFEREL
pH 4.0, 37°C
0.041
AFPLEFFREL
pH 4.0, 37°C
0.079
AFPLEFIREL
pH 4.0, 37°C
0.18
AFPLEFKREL
pH 4.0, 37°C
0.0056 - 0.007
Abz-A-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
0.004
Abz-A-A-F-F-A-A-p-nitroanilide
-
hydrolysis by calf chymosin
0.0022 - 0.018
Abz-A-A-F-F-A-N-(2,4-dinitrophenyl)-ethylenediamine
0.003
Abz-A-A-F-F-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by calf chymosin
0.0018
Abz-A-A-F-F-p-nitroanilide
-
hydrolysis by calf chymosin
0.0018 - 0.0078
Abz-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
0.02 - 0.89
kappa-casein
-
0.333
Leu-Ser-Phe(NO2)-Nle-Ala-Leu-O-methyl ester
-
point mutant G243D
1.11
Leu-Ser-Phe(NO2)-Nle-Ala-Leu-Ome
-
point mutant A115T
0.85
Leu-Ser-Phe-Met-Ala-Ile-O-methyl ester
-
-
6.9
Leu-Ser-Phe-Met-Ala-O-methyl ester
-
-
0.34
Pro-His-Leu-Ser-Phe-Met-Ala-Ile-O-methyl ester
-
-
8.5
Ser-Phe-Met-Ala-Ile-O-methyl ester
-
-
0.165
undecapeptide analogue to chymosin sensitive region of bovine kappa casein
-
-
-
0.134
undecapeptide analogue to chymosin sensitive region of camel kappa casein
-
-
-
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2600
AFPLEFEREL
pH 4.0, 37°C
290
AFPLEFFREL
pH 4.0, 37°C
427
AFPLEFIREL
pH 4.0, 37°C
29
AFPLEFKREL
pH 4.0, 37°C
0.12 - 1.6
Abz-A-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
1.4
Abz-A-A-F-F-A-A-p-nitroanilide
-
hydrolysis by calf chymosin
0.02 - 0.8
Abz-A-A-F-F-A-N-(2,4-dinitrophenyl)-ethylenediamine
0.27
Abz-A-A-F-F-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by calf chymosin
0.48
Abz-A-A-F-F-p-nitroanilide
-
hydrolysis by calf chymosin
0.06 - 0.14
Abz-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
44.3
undecapeptide analogue to chymosin sensitive region of bovine kappa casein
-
-
-
4.3
undecapeptide analogue to chymosin sensitive region of camel kappa casein
-
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
790
AFPLEFEREL
pH 4.0, 37°C
7100
AFPLEFFREL
pH 4.0, 37°C
5400
AFPLEFIREL
pH 4.0, 37°C
160
AFPLEFKREL
pH 4.0, 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.7
-
isoform chymosin B
4.2
-
isoform chymosin A
4.9
-
for proteolysis of undecapeptide analogue to chymosin sensitive region of the bovine kappa casein
6.5
-
pH optimum for the Tol-A-III-chymosin fusion protein
6.6
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2 - 6
-
-
2 - 6.7
-
-
3 - 10
-
pH 3: about 80% of maximum activity, pH 10: about 20% of maximum activity
4.5
-
using alpha- and beta-casein as substrate
4.6 - 5.6
-
-
5 - 6.9
-
recombinant chymosin shows coagulant activity from pH 5.0 to 6.9
5 - 9
-
the Tol-A-III-chymosin fusion protein shows more than 60% activity between pH 5.0 and 9.0
5.5
-
using kappa-casein as substrate
5.5 - 6
-
100% activity at pH 5.5, about 90% activity at pH 6.0, about 30% activity at pH 6.5, the enzyme almost completely loses its activity at pH 7.0
5.6
-
-
5.8
-
using whole casein as substrate
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
assay at
30 - 50
-
-
35 - 40
-
-
40
-
temperature optimum for the Tol-A-III-chymosin fusion protein
42
-
for proteolysis of the undecapeptide analogue to chymosin sensitive region of bovine kappa casein
45
-
at pH 6.6
50
-
-
56
-
for proteolysis of the undecapeptide analogue to chymosin sensitive region of camel kappa casein
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 60
-
20°C: about 25% of maximum activity, 60°C: about 50% of maximum activity
25 - 50
-
recombinant chymosin shows coagulant activity from 25 to 50°C
30 - 55
-
the Tol-A-III-chymosin fusion protein shows high activity between 30 and 55°C
5 - 52.5
-
-
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.55
-
isoelectric focusing
4.6
-
recombinant chymosin
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
additional information
-
rennet from abomasal tissue from a local indigenous breed
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
-
chymosin, an aspartic protease, is the main enzymatic component of calf rennet
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
CHYM_BOVIN
381
0
42180
Swiss-Prot
Secretory Pathway (Reliability: 2)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30700
-
amino acid composition
33000
-
gel filtration
35600
-
when the proenzyme is brought to pH 4.2, removal of 42 N-terminal residues results in an active form of chymosin of 35600 Da
36000
36300
-
two-dimensional electrophoresis technique
36500
-
sedimentation, diffusion
40000
-
x * 40000, SDS-PAGE
additional information
-
-
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * about 40000, mass spectrometry
additional information
-
? * 36000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
glycosylation at Asn291 significantly decreases enzyme activity
glycoprotein
-
-
additional information
-
insertion of a glycosylation site (S351T), glycosylation has significant inhibitory effect but this can be circumvented by deglycosylation with endoglucosidase H
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
unglycosylated chymosin, vapor diffusion method, using 100 mM NaH2PO4 pH 5.5, 1.5 M NaCl
loop exchange mutant
-
modeling of chymosin in complex with residues 97-112 of bovine kappa-casein. Substrate binds in an extended conformation with charged residues on either side of the scissile bond playing an important role in stabilizing the binding pose. Substrate residues Lys111 and Lys112 bind to the N-terminal domain of chymosin displacing a conserved water molecule. A cluster of histidine and proline residues, His98-Pro99-His100-Pro101-His102, in kappa-casein binds to the C-terminal domain of the protein, where neighboring conserved arginine residue Arg97 is important for stabilizing the binding pose. The catalytic site including the catalytic water molecule is stable in the starting conformation of the general acid/base catalytic mechanism for 18 ns of molecular dynamics simulations
-
point mutants
-
prochymosin mutant
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A115T
-
reduction of Km compared to wild-type
D304A
-
pH optima shift
D304A/T218A
G243D
-
increase of Km compared to wild-type
G244D
-
pH optima shift
Q288K
-
-
S351T
-
insertion of a glycosylation site in the linker region between chymosin and glucoamylase, strongly improves enzyme secretion
T218A
-
pH optima shift
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2
-
-
36882
2 - 6
-
under incubation at 37°C for 24 and 48 h, loss of activity at pH 3.5 and pH 7.0, trangenic chymosin and recombinant chymosin are inactivated slightly faster than the native enzyme
667763
2 - 6.5
-
crystalline chymosin exhibits optimum stability at pH between 5.3 and 6.3, the enzyme is moderately stable at pH 2.0, but the enzyme is unstable at pH around 3.5 and above 6.5, clotting activity is not observed at pH 7.0
708236
3.5
-
rapid loss of activity
36882
5.3 - 6.3
-
highest stability
36882
7
-
rapid loss of activity above
36882
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 50
-
transgenic chymosin loses activity slightly faster than abomasal enzyme and recombinant enzyme
35 - 55
-
the enzyme activity remains stable after 30 min at 35-55°C and drops to about 20% after 30 min at 60°C
45
-
about 25% loss of activity after 60 min
5
-
very stable below, suspension of crystals
50
-
about 80% loss of activity after 30 min
50 - 60
-
chymosin is stable up to 50°C and a relative milk-clotting activity of 50% is recorded when the temperature is raised to 60°C
55
-
95% loss of activity after 10 min
56
-
chymosin-mediated hydrolysis of alphas1 casein is slower in cheeses treated at 56°C
60 - 100
-
incubation at 60°C for 2 min 30 s decreases the mil clotting activity of the enzyme. A 2 min heating at 80 and 100°C totally inactivates the enzyme
75
-
it is not possible to coagulate milk with chymosin at 75°C due to its loss of activity
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
urea
-
chymosin loses 50% of its activity after incubation in 4.6 M urea at 37°C for 30 min
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, at pH 3.0 and 6.2, 3 days, maximum stability
-
very stable below 5°C, suspension of crystals
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
phenyl Superose column chromatography
by affinity chromatography on a bacitracin-Sepharose column
-
DEAE-cellulose column chromatography, gel filtration
-
extraction by application of ultrasound
-
gel filtration and anion-exchange chromatography
-
Ni-NTA affinity column chromatography
-
purification by partitioning between aqueous two-phase systems
-
ultrafiltration and Superdex 75 gel filtration
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed as inclusion bodies in Escherichia coli
-
expressed in Escherichia coli and in Saccharomyces cerevisiae
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Nicotiana tabacum cultivar Petite Havana
-
expressed in Pichia pastoris strain GS115
-
expressed in Pichia pastoris strain KM71H
-
expression in Aspergillus niger
-
expression in Kluyveromyces lactis
-
loop exchange mutant expressed in Trichoderma reesei
-
point mutants expressed in Escherichia coli and Trichoderma reesei
-
prochymosin fusion product expressed in Escherichia coli
-
S351T mutant of prochymosin as fusion protein with the Aspergillus niger enzyme glucoamylase in Aspergillus niger var. awamori, insertion of a short peptide linker between the proteins with the sequence TDNST
-
the enzyme as a fusion with Tol-A-III protein is expressed in Escherichia coli BL21 (DE3) cells
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
oxidative refolding of solubilized enzyme obtained from inclusion bodies at pH 2 to obtain autoconversion of prochymosin to active chymosin
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
food industry
the enzyme is used industrially in cheese production
food industry
synthesis
-
use of chymosin to cleave a pro-chymosin derived fusion tag releasing native target proteins. After modification of the pro-chymosin fusion tag chymosin can remove this tag at more neutral pH 6.2, less prone to compromise the integrity of target proteins. Chymosin produces intact native target protein both at the level of small and large-scale preparations
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Malak, C.A.A.; El Adab, I.F.G.A.; Vukashinovic, V.; Zalunin, I.A.; Timokhina, E.A.; Lavrenova, G.I.; Stepanov, V.M.
Buffalo (Bos buffali L.) chymosin purification and properties
Comp. Biochem. Physiol. B
113
57-62
1996
Bos taurus
Manually annotated by BRENDA team
Moir, D.; Mao, J.I.; Schumm, J.W.; Vovis, G.F.; Alford, B.L.; Taunton-Rigby, A.
Molecular cloning and characterization of double-stranded cDNA coding for bovine chymosin
Gene
19
127-138
1982
Bos taurus
Manually annotated by BRENDA team
Beppu, T.
The cloning and expression of chymosin (rennin) genes in microorganisms
Trends Biotechnol.
1
85-89
1983
Bos taurus
-
Manually annotated by BRENDA team
Gustchina, E.; Rumsh, L.; Ginodman, L.; Majer, P.; Andreeva, N.
Post X-ray crystallographic studies of chymosin: the existence of two structural forms and the regulation of activity by the interaction with the histidine-proline cluster of kappa-casein
FEBS Lett.
379
60-62
1996
Bos taurus
Manually annotated by BRENDA team
Kleinert, T.; Lange, I.; Roesicke, B.; Honig, A.; Schleusener, R.
Characterization and preparation of chymosin from calf rennet samples by means of isoelectric focusing
Acta Biotechnol.
8
367-375
1988
Bos taurus
-
Manually annotated by BRENDA team
Zayas, J.F.
Properties and quality characteristics of rennin extracted by ultrasound
Biotechnol. Bioeng.
29
969-975
1987
Bos taurus
Manually annotated by BRENDA team
Kawaguchi, Y.; Kosugi, S.; Sasaki, K.; Uozumi, T.; Beppu, T.
Production of chymosin in Escherichia coli cells and its enzymatic properties
Agric. Biol. Chem.
51
1871-1877
1987
Bos taurus
-
Manually annotated by BRENDA team
McCaman, M.T.; Andrews, W.H.; Files, J.G.
Enzymatic properties and processing of bovine prochymosin synthezised in Escherichia coli
J. Biotechnol.
2
177-190
1985
Bos taurus
-
Manually annotated by BRENDA team
Raap, J.; Kerling, K.E.T.; Vreeman, H.J.; Visser, S.
Peptide substrates for chymosin (rennin): conformational studies of kappa-casein and some kappa-casein-related oligopeptides by circular dichroism and secondary structure prediction
Arch. Biochem. Biophys.
221
117-124
1983
Bos taurus
Manually annotated by BRENDA team
Miyoshi, M.; Yoon, C.H.; Ibuki, F.; Kanamori, M.
The characterization of rennin action on kappa-casein using CM-cellulose
Agric. Biol. Chem.
40
347-352
1976
Bos taurus
-
Manually annotated by BRENDA team
Chang, W.J.; Takahashi, K.
The structure and function of acid proteases. III. Isolation and characterization of the active-site peptides from bovine rennin
J. Biochem.
76
467-474
1974
Bos taurus
Manually annotated by BRENDA team
Williams, M.G.; Wilsher, J.; Nugent, P.; Mills, A.; Dhanaraj, V.; Fabry, M.; Sedlacek, J.; Uusitalo, J.M.; Penttila, M.E.; Pitts, J.E.; Blundell, T.L.
Mutagenesis, biochemical characterization and X-ray structural analysis of point mutants of bovine chymosin
Protein Eng.
10
991-997
1997
Bos taurus
Manually annotated by BRENDA team
Foltmann, B.
Prochymosin and chymosin (prorennin and rennin)
Methods Enzymol.
19
421-436
1970
Bos taurus
-
Manually annotated by BRENDA team
Azuma, N.; Kaminogawa, S.; Yamauchi, K.
Properties of glycomacropeptide and para-kappa-casein derived from human kappa-casein and comparison of human and bovine kappa-casein as to susceptibility to chymosin and pepsin
Agric. Biol. Chem.
48
2025-2031
1984
Bos taurus, Homo sapiens
-
Manually annotated by BRENDA team
Pitts, J.E.; Quinn, D.; Uusitalo, J.; Penttilae, M.
Protein engineering of chymosin and expression in Trichoderma reesei
Food Biotechnol.
19
663-666
1991
Bos taurus
Manually annotated by BRENDA team
Strop, P.; Sedlacek, J.; Stys, J.; Kaderabkova, Z.; Blaha, I.; Pavlickova, L.; Pohl, J.; Fabry, M.; Kostka, V.; Newman, M.; Frazao, C.; Shearer, A.; Tickle, I.J.; Blundell, T.L.
Engineering enzyme subsite specificity: preparation, kinetic characterization, and X-ray analysis at 2.0-A resolution of Val111Phe site-mutated calf chymosin
Biochemistry
29
9863-9871
1990
Bos taurus
Manually annotated by BRENDA team
Pitts, J.E.; Uusitalo, J.M.; Mantafounis, D.; Nugent, P.G.; Quinn, D.D.; Orprayoon, P.; Penttilae, M.E.
Expression and characterisation of chymosin pH optima mutants produced in Trichoderma reesei
J. Biotechnol.
28
69-83
1993
Bos taurus
Manually annotated by BRENDA team
Nugent, P.G.; Albert, A.; Orprayoon, P.; Wilsher, J.; Pitts, J.E.; Blundell, T.L.; Dhanaraj, V.
Protein engineering loops in aspartic proteinases: site-directed mutagenesis, biochemical characterization and X-ray analysis of chymosin with a replaced loop from rhizopuspepsin
Protein Eng.
9
885-893
1996
Bos taurus
Manually annotated by BRENDA team
Mantafounis, D.; Pitts, J.
Protein engineering of chymosin; modification of the optimum pH of enzyme catalysis
Protein Eng.
3
605-609
1990
Bos taurus
Manually annotated by BRENDA team
Gustchina, E.A.; Majer, P.; Rumsh, L.D.; Ginodman, L.M.; Andreeva, N.S.
Post X-ray crystallographic studies of chymosin specificity. The role of histidine-proline cluster of kappa-casein in catalytic reactions
Adv. Exp. Med. Biol.
436
179-184
1998
Bos taurus
Manually annotated by BRENDA team
Kappeler, S.R.; van den Brink, H.J.; Rahbek-Nielsen, H.; Farah, Z.; Puhan, Z.; Hansen, E.B.; Johansen, E.
Characterization of recombinant camel chymosin reveals superior properties for the coagulation of bovine and camel milk
Biochem. Biophys. Res. Commun.
342
647-654
2006
Bos taurus, Camelus dromedarius
Manually annotated by BRENDA team
Starovoitova, V.V.; Velichko, T.I.; Baratova, L.A.; Filippova, I.Y.; Lavrenova, G.I.
A comparative study of functional properties of calf chymosin and its recombinant forms
Biochemistry (Moscow)
71
320-324
2006
Bos taurus
Manually annotated by BRENDA team
Huppertz, T.; Uniacke, T.; Kelly, A.L.; Fox, P.F.
Inhibition of the proteolytic activity of indigenous plasmin or exogenous chymosin and pepsin in bovine milk by blood serum
Int. dairy J.
16
691-696
2006
Bos taurus
Manually annotated by BRENDA team
Moschopoulou, E.E.; Kandarakis, I.G.; Alichanidis, E.; Anifantakis, E.M.
Purification and characterization of chymosin and pepsin from kid
J. Dairy Res.
73
49-57
2006
Bos taurus
Manually annotated by BRENDA team
Molle, D.; Jean, K.; Guyomarch, F.
Chymosin sensitivity of the heat-induced serum protein aggregates isolated from skim milk
Int. Dairy J.
16
1435-1441
2006
Bos taurus
Manually annotated by BRENDA team
Bansal, N.; Fox, P.F.; McSweeney, P.L.
Aggregation of rennet-altered casein micelles at low temperatures
J. Agric. Food Chem.
55
3120-3126
2007
Bos taurus
Manually annotated by BRENDA team
Renan, M.; Guyomarch, F.; Chatriot, M.; Gamerre, V.; Famelart, M.H.
Limited enzymatic treatment of skim milk using chymosin affects the micelle/serum distribution of the heat-induced whey protein/kappa-casein aggregates
J. Agric. Food Chem.
55
6736-6745
2007
Bos taurus
Manually annotated by BRENDA team
van den Brink, H.J.; Petersen, S.G.; Rahbek-Nielsen, H.; Hellmuth, K.; Harboe, M.
Increased production of chymosin by glycosylation
J. Biotechnol.
125
304-310
2006
Bos taurus
Manually annotated by BRENDA team
Reh, G.; Spelzini, D.; Tubio, G.; Pico, G.; Farruggia, B.
Partition features and renaturation enhancement of chymosin in aqueous two-phase systems
J. Chromatogr. B
860
98-105
2007
Bos taurus
Manually annotated by BRENDA team
Palmer, D.S.; Christensen, A.U.; Srensen, J.; Celik, L.; Qvist, K.B.; Schiott, B.
Bovine chymosin: a computational study of recognition and binding of bovine kappa-casein
Biochemistry
49
2563-2573
2010
Bos taurus
Manually annotated by BRENDA team
Hidalgo, M.E.; Pires, M.S.; Risso, P.H.
A study on bovine kappa-casein aggregation after the enzymatic action of chymosin
Colloids Surf. B Biointerfaces
76
556-563
2010
Bos taurus
Manually annotated by BRENDA team
Kumar, A.; Grover, S.; Sharma, J.; Batish, V.K.
Chymosin and other milk coagulants: sources and biotechnological interventions
Crit. Rev. Biotechnol.
30
243-258
2010
Bos taurus
Manually annotated by BRENDA team
Kageyama, H.; Ueda, H.; Tezuka, T.; Ogasawara, A.; Narita, Y.; Kageyama, T.; Ichinose, M.
Differences in the P1' substrate specificities of pepsin A and chymosin
J. Biochem.
147
167-174
2010
Bos taurus (P00794), Bos taurus
Manually annotated by BRENDA team
Justesen, S.F.; Lamberth, K.; Nielsen, L.L.; Schafer-Nielsen, C.; Buus, S.
Recombinant chymosin used for exact and complete removal of a prochymosin derived fusion tag releasing intact native target protein
Protein Sci.
18
1023-1032
2009
Bos taurus
Manually annotated by BRENDA team
Sorensen, J.; Palmer, D.S.; Qvist, K.B.; Schiott, B.
Initial stage of cheese production: a molecular modeling study of bovine and camel chymosin complexed with peptides from the chymosin-sensitive region of kappa-casein
J. Agric. Food Chem.
59
5636-5647
2011
Bos taurus (P00794), Bos taurus, Camelus dromedarius (Q9GK11)
Manually annotated by BRENDA team
Langholm Jensen, J.; Molgaard, A.; Navarro Poulsen, J.C.; Harboe, M.K.; Simonsen, J.B.; Lorentzen, A.M.; Hjerno, K.; van den Brink, J.M.; Qvist, K.B.; Larsen, S.
Camel and bovine chymosin: the relationship between their structures and cheese-making properties
Acta Crystallogr. Sect. D
69
901-913
2013
Bos taurus (P00794), Bos taurus, Camelus dromedarius (Q9GK11)
Manually annotated by BRENDA team
Bijl, E.; van Valenberg, H.; Sikkes, S.; Jumelet, S.; Sala, G.; Olieman, K.; van Hooijdonk, T.; Huppertz, T.
Chymosin-induced hydrolysis of caseins: Influence of degree of phosphorylation of alpha-s1-casein and genetic variants of beta-casein
Int. Dairy J.
39
215-221
2014
Bos taurus
-
Manually annotated by BRENDA team
Moller, K.K.; Rattray, F.P.; Sorensen, J.C.; Ardoe, Y.
Comparison of the hydrolysis of bovine kappa-casein by camel and bovine chymosin: a kinetic and specificity study
J. Agric. Food Chem.
60
5454-5460
2012
Bos taurus, Camelus dromedarius
Manually annotated by BRENDA team
Costabel, L.M.; Bergamini, C.V.; Pozza, L.; Cuffia, F.; Candioti, M.C.; Hynes, E.
Influence of chymosin type and curd scalding temperature on proteolysis of hard cooked cheeses
J. Dairy Res.
85
375-384
2015
Bos taurus
Manually annotated by BRENDA team
Jensen, J.L.; Jacobsen, J.; Moss, M.L.; Rasmussen, F.; Qvist, K.B.; Larsen, S.; van den Brink, J.M.
The function of the milk-clotting enzymes bovine and camel chymosin studied by a fluorescence resonance energy transfer assay
J. Dairy Sci.
98
2853-2860
2015
Bos taurus, Camelus dromedarius
Manually annotated by BRENDA team
Noseda, D.G.; Blasco, M.; Recupero, M.; Galvagno, M.A.
Bioprocess and downstream optimization of recombinant bovine chymosin B in Pichia (Komagataella) pastoris under methanol-inducible AOXI promoter
Protein Expr. Purif.
104
85-91
2014
Bos taurus
Manually annotated by BRENDA team
Belenkaya, S.; Rudometov, A.; Shcherbakov, D.; Balabova, D.; Kriger, A.; Belov, A.; Koval, A.; Elchaninov, V.
Biochemical properties of recombinant chymosin in alpaca (Vicugna pacos L.)
Appl. Biochem. Microbiol.
54
569-576
2018
Bos taurus, Vicugna pacos
-
Manually annotated by BRENDA team
Rayanatou, I.A.; Mahamadou, E.G.; Garric, G.; Harel-Oger, M.; Leduc, A.; Jardin, J.; Briard-Bion, V.; Cauty, C.; Adakal, H.; Grongnet, J.F.; Gaucheron, F.
Physico-chemical characterization of dairy gel obtained by a proteolytic extract from Calotropis procera - A comparison with chymosin
Food Chem.
232
405-412
2017
Bos taurus
Manually annotated by BRENDA team
Wei, Z.Y.; Zhang, Y.Y.; Wang, Y.P.; Fan, M.X.; Zhong, X.F.; Xu, N.; Lin, F.; Xing, S.C.
Production of bioactive recombinant bovine chymosin in tobacco plants
Int. J. Mol. Sci.
17
E624
2016
Bos taurus
Manually annotated by BRENDA team
Rolet-Repecaud, O.; Arnould, C.; Dupont, D.; Gavoye, S.; Beuvier, E.; Achilleos, C.
Development and evaluation of a monoclonal antibody-based inhibition ELISA for the quantification of chymosin in solution
J. Agric. Food Chem.
63
4799-4804
2015
Bos taurus
Manually annotated by BRENDA team
Ansari, S.M.; Coletta, A.; Kirkeby Skeby, K.; Sorensen, J.; Schiott, B.; Palmer, D.S.
Allosteric-activation mechanism of bovine chymosin revealed by bias-exchange metadynamics and molecular dynamics simulations
J. Phys. Chem. B
120
10453-10462
2016
Bos taurus (P00794), Bos taurus
Manually annotated by BRENDA team
Leite Junior, B.; Tribst, A.; Cristianini, M.
The effect of high pressure processing on recombinant chymosin, bovine rennet and porcine pepsin Influence on the proteolytic and milk-clotting activities and on milk-clotting characteristics
LWT-Food Sci. Technol.
76
351-360
2017
Bos taurus
-
Manually annotated by BRENDA team
Espinoza-Molina, J.A.; Acosta-Muniz, C.H.; Sepulveda, D.R.; Zamudio-Flores, P.B.; Rios-Velasco, C.
Codon optimization of the "Bos taurus Chymosin" gene for the production of recombinant chymosin in Pichia pastoris
Mol. Biotechnol.
58
657-664
2016
Bos taurus
Manually annotated by BRENDA team
Ulusu, Y.; Sentuerk, S.B.; Kudug, H.; Goekce, I.
Expression, purification, and characterization of bovine chymosin enzyme using an inducible pTOLT system
Prep. Biochem. Biotechnol.
46
596-601
2016
Bos taurus
Manually annotated by BRENDA team
Ansari, S.M.; Sorensen, J.; Schiott, B.; Palmer, D.S.
On the effect of mutations in bovine or camel chymosin on the thermodynamics of binding kappa-caseins
Proteins
86
75-87
2018
Bos taurus (P00794), Bos taurus, Camelus dromedarius (Q9GK11)
Manually annotated by BRENDA team