Information on EC 3.4.23.4 - chymosin

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The expected taxonomic range for this enzyme is: Opisthokonta

EC NUMBER
COMMENTARY
3.4.23.4
-
RECOMMENDED NAME
GeneOntology No.
chymosin
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
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
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
endopeptidase; peptides, endopeptidase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
chymase
-
-
-
-
chymosin
Q9N1p5
-
chymosin A
-
allelic form
chymosin B
-
allelic form
Preprorennin
-
-
-
-
prochymosin
-
inactive
prochymosin
-
cDNA predicted to code for 365 amino acids with a 42 amino acids long pro-region
prochymosin
Q5VK60
inactive precursor
rennin
-
-
-
-
EC 3.4.4.3
-
-
formerly
-
additional information
Q5VK60
acid protease, major component of rennet (milk clotting enzyme)
CAS REGISTRY NUMBER
COMMENTARY
9001-98-3
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
similarar enzyme
-
-
Manually annotated by BRENDA team
chymosin derived as commercial enzyme preparation (Maxiren) from a genetically modified yeast Kluyveromyces lactic
-
-
Manually annotated by BRENDA team
use of a synthetic gene that encodes a protein identical to bovine prochymosin
-
-
Manually annotated by BRENDA team
synthetic construct of prochymosin mRNA; enzyme secretion is maximal during the first days after birth, enzyme level declines thereafter and chymosin in replaced by pepsin
UniProt
Manually annotated by BRENDA team
similar enzyme
-
-
Manually annotated by BRENDA team
similar enzyme
-
-
Manually annotated by BRENDA team
similar enzyme
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
chymosin, an aspartic protease, is the main enzymatic component of calf rennet
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
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
-
-
-
-
?
acid denatured hemoglobin + H2O
?
show the reaction diagram
-
-
-
-
?
AFPLEFEREL + H2O
AFPLEF + EREL
show the reaction diagram
P00794
modified peptide substrate based on residues 165-174 of proopiomelanocortin
-
-
?
AFPLEFFREL + H2O
AFPLEF + FREL
show the reaction diagram
P00794
modified peptide substrate based on residues 165-174 of proopiomelanocortin
-
-
?
AFPLEFIREL + H2O
AFPLEF + IREL
show the reaction diagram
P00794
modified peptide substrate based on residues 165-174 of proopiomelanocortin
-
-
?
AFPLEFKREL + H2O
AFPLEF + KREL
show the reaction diagram
P00794
modified peptide substrate based on residues 165-174 of proopiomelanocortin
-
-
?
alpha-casein + H2O
?
show the reaction diagram
-
-
-
-
?
alphaS-casein + H2O
?
show the reaction diagram
-
-
-
-
?
basic FGF 110-118 + H2O
?
show the reaction diagram
-
parent peptide and Leu115 and Lys115 variants respectively as substrate
-
-
?
beta-casein + H2O
?
show the reaction diagram
-
-
-
-
?
beta-casein + H2O
?
show the reaction diagram
-
-
-
-
?
beta-casein + H2O
?
show the reaction diagram
-
occurs during the long period of renneting
-
-
?
beta-chain of proteolytic insulin + H2O
?
show the reaction diagram
-
general proteolytic activity
-
-
?
bovine kappa-casein + H2O
?
show the reaction diagram
-
kappa-casein samples are a mixture of monomers and aggregates at room temperature and pH 7.2, and that heating produces extensive kappa-casein aggregation.The initial polymerization or association state of kappa-casein affects on the aggregation stage after the enzymatic action of chymosin. Sucrose and lactose also affect the aggregation of proteolized particles of kappa-chymosin
-
-
?
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
-
-
-
-
?
dynorphin A 1-7e + H2O
?
show the reaction diagram
-
Ala3Phe7 and ILe3Lys5Phe7 variants respectively as substrate
-
-
?
fluorescein thiocarbamoyl-kappa-casein + H2O
?
show the reaction diagram
-, Q9N1p5
-
-
-
?
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
-
part of the bovine kappa-casein sequence
-
?
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
-
part of the bovine kappa-casein sequence
-
?
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
p-kappa-casein + glycomacropeptide
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
?
kappa-casein + H2O
p-kappa-casein + glycomacropeptide
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
?
kappa-casein + H2O
p-kappa-casein + glycomacropeptide
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
?
kappa-casein + H2O
p-kappa-casein + glycomacropeptide
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
?
kappa-casein + H2O
p-kappa-casein + glycomacropeptide
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
?
kappa-casein + H2O
p-kappa-casein + glycomacropeptide
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
p-kappa-casein i.e. N-terminal fraction, glycomacropeptide i.e. C-terminal fraction
?
kappa-casein + H2O
p-kappa-casein + glycomacropeptide
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
p-kappa-casein i.e. N-terminal fraction, glycomacropeptide i.e. C-terminal fraction
?
kappa-casein + H2O
p-kappa-casein + glycomacropeptide
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
p-kappa-casein i.e. N-terminal fraction, glycomacropeptide i.e. C-terminal fraction
?
kappa-casein + H2O
?
show the reaction diagram
Q5VK60
-
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-, Q9N1p5
-
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
-
-
-
?
kappa-casein + H2O
?
show the reaction diagram
Q9GK11
-
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond between phenylalanine 105 and methionine 106
-
-
?
kappa-casein + H2O
?
show the reaction diagram
Q5VK60
cleaves a single bond between phenylalanine 105 and methionine 106
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
involved in milk clotting
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
involved in milk clotting
-
-
?
kappa-casein + H2O
casein macropeptide + para-kappa-casein
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
-
-
?
kappa-casein + H2O
para-kappa-casein + macropeptide
show the reaction diagram
-
kappa-casein is the primary substrate for the chymosin action
-
-
?
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
-
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
-
part of the bovine kappa-casein sequence
-
?
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-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
-
-
-
?
NT/NMN 142-151 + H2O
?
show the reaction diagram
-
parent peptide, Glu148 and Phe148 variants respectively as substrate
-
-
?
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
-
-
-
?
POm C165-174 + H2O
?
show the reaction diagram
-
parent peptide and Glu171 variant respectively as substrate
-
-
?
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
-
?
Substance P + H2O
?
show the reaction diagram
-
parent peptide and Lys8 variant respectively as substrate
-
-
?
undecapeptide analogue to chymosin sensitive region of bovine kappa casein
?
show the reaction diagram
-
synthetic substrate
-
-
?
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
-
-
?
undecapeptide analogue to chymosin sensitive region of camel kappa-casein
?
show the reaction diagram
-
synthetic substrate
-
-
?
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 + YE
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
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond in kappa-casein between phenylalanine 105 and methionine 106
-
-
-
kappa-casein + H2O
?
show the reaction diagram
-
cleaves a single bond between phenylalanine 105 and methionine 106
-
-
?
kappa-casein + H2O
?
show the reaction diagram
Q5VK60
cleaves a single bond between phenylalanine 105 and methionine 106
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
involved in milk clotting
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
involved in milk clotting
-
-
?
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
-
-
?
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
stimulation of milk clotting activity
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
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
-
Diazoacetylnorleucine methyl ester
-
in presence of Cu2+
Pepstatin
-
-
Pepstatin
-
inhibits recombinant and transgenic chymosin less efficiently than the native enzyme
pepstatin A
-
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
His-Pro-His-Pro-His-NH2
-
stimulation, part of kappa-casein
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0056
-
Abz-A-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by recombinant chymosin; hydrolysis by transgenic chymosin
0.007
-
Abz-A-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by calf chymosin
0.004
-
Abz-A-A-F-F-A-A-p-nitroanilide
-
hydrolysis by calf chymosin
0.0022
-
Abz-A-A-F-F-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by calf chymosin
0.0035
-
Abz-A-A-F-F-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by recombinant chymosin
0.018
-
Abz-A-A-F-F-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by transgenic chymosin
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
-
Abz-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by calf chymosin
0.0021
-
Abz-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by recombinant chymosin
0.0078
-
Abz-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by transgenic chymosin
3.3
-
AFPLEFEREL
-
pH 4.0, 37C
0.041
-
AFPLEFFREL
-
pH 4.0, 37C
0.079
-
AFPLEFIREL
-
pH 4.0, 37C
0.18
-
AFPLEFKREL
-
pH 4.0, 37C
0.01223
-
kappa-casein
-, Q9N1p5
-
-
0.02
-
kappa-casein
-
point mutant G343D
-
0.066
-
kappa-casein
-
-
-
0.106
-
kappa-casein
-
-
-
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
-
-
0.016
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage L147-F148) by D289M/Q298L mutant chymosin
0.024
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage F147-R148) by D289M/Q298L mutant chymosin
0.028
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage F147-R148) by Asp289Met mutant chymosin
0.029
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage L147-F148) by Gln298Leu mutant chymosin
0.033
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage F147-R148) by Gln298Leu mutant chymosin
0.046
-
NT/NMN 142-151
-
hydrolysis of the Glu148 variant by D289M/Q298L mutant chymosin
0.054
-
NT/NMN 142-151
-
hydrolysis of the Glu148 variant by Asp289Met mutant chymosin; hydrolysis of the Phe148 variant (cleavage L147-F148) by Asp289Met mutant chymosin
0.059
-
NT/NMN 142-151
-
hydrolysis of the parent peptide by wild type chymosin
0.062
-
NT/NMN 142-151
-
hydrolysis of the Glu148 variant by Gln298Leu mutant chymosin
0.072
-
NT/NMN 142-151
-
hydrolysis of the parent peptide by Asp289Met mutant chymosin
0.083
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage L147-F148) by wild type chymosin
0.087
-
NT/NMN 142-151
-
hydrolysis of the parent peptide by D289M/Q298L mutant chymosin
0.095
-
NT/NMN 142-151
-
hydrolysis of the parent peptide by Gln298Leu mutant chymosin
0.105
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage F147-R148) by wild type chymosin
0.111
-
NT/NMN 142-151
-
hydrolysis of the Glu148 variant by wild type chymosin
0.065
-
POm C165-174
-
hydrolysis of the Glu171 variant by Asp289Met mutant chymosin
0.114
-
POm C165-174
-
hydrolysis of the Glu171 variant by D289M/Q298L mutant chymosin
0.229
-
POm C165-174
-
hydrolysis of the parent peptide by Gln298Leu mutant chymosin
0.25
-
POm C165-174
-
hydrolysis of the parent peptide by D289M/Q298L mutant chymosin
0.288
-
POm C165-174
-
hydrolysis of the parent peptide by Asp289Met mutant chymosin
0.304
-
POm C165-174
-
hydrolysis of the Glu171 variant by wild type chymosin
0.375
-
POm C165-174
-
hydrolysis of the parent peptide by wild type chymosin
0.463
-
POm C165-174
-
hydrolysis of the Glu171 variant by Gln298Leu mutant chymosin
0.34
-
Pro-His-Leu-Ser-Phe-Met-Ala-Ile-O-methyl ester
-
-
8.5
-
Ser-Phe-Met-Ala-Ile-O-methyl ester
-
-
0.077
-
undecapeptide analogue to chymosin sensitive region of bovine kappa casein
-
-
-
0.165
-
undecapeptide analogue to chymosin sensitive region of bovine kappa casein
-
-
-
0.056
-
undecapeptide analogue to chymosin sensitive region of camel kappa casein
-
-
-
0.134
-
undecapeptide analogue to chymosin sensitive region of camel kappa casein
-
-
-
6.9
-
Leu-Ser-Phe-Met-Ala-O-methyl ester
-
-
additional information
-
additional information
-
Km of kappa-casein related peptides
-
additional information
-
additional information
-
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.12
-
Abz-A-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by recombinant chymosin
0.16
-
Abz-A-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by transgenic chymosin
1.6
-
Abz-A-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by calf chymosin
1.4
-
Abz-A-A-F-F-A-A-p-nitroanilide
-
hydrolysis by calf chymosin
0.02
-
Abz-A-A-F-F-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by transgenic chymosin
0.11
-
Abz-A-A-F-F-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by recombinant chymosin
0.8
-
Abz-A-A-F-F-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by calf chymosin
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
-
Abz-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by transgenic chymosin or recombinat chymosin
0.14
-
Abz-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
-
hydrolysis by calf chymosin
2600
-
AFPLEFEREL
-
pH 4.0, 37C
290
-
AFPLEFFREL
-
pH 4.0, 37C
427
-
AFPLEFIREL
-
pH 4.0, 37C
29
-
AFPLEFKREL
-
pH 4.0, 37C
2
8
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage L147-F148) by wild type chymosin
12
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage L147-F148) by Gln298Leu mutant chymosin
16
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage L147-F148) by D289M/Q298L mutant chymosin
17
-
NT/NMN 142-151
-
hydrolysis of the Glu148 variant by Gln298Leu mutant chymosin; hydrolysis of the Phe148 variant (cleavage F147-R148) by D289M/Q298L mutant chymosin
22
-
NT/NMN 142-151
-
hydrolysis of the Glu148 variant by D289M/Q298L mutant chymosin
27
-
NT/NMN 142-151
-
hydrolysis of the parent peptide by D289M/Q298L mutant chymosin
30
-
NT/NMN 142-151
-
hydrolysis of the parent peptide by Gln298Leu mutant chymosin
32
-
NT/NMN 142-151
-
hydrolysis of the Glu148 variant by Asp289Met mutant chymosin
35
-
NT/NMN 142-151
-
hydrolysis of the parent peptide by Asp289Met mutant chymosin
39
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage L147-F148) by Asp289Met mutant chymosin
41
-
NT/NMN 142-151
-
hydrolysis of the parent peptide by wild type chymosin
42
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage F147-R148) by Asp289Met mutant chymosin
46
-
NT/NMN 142-151
-
hydrolysis of the Glu148 variant by wild type chymosin
49
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage F147-R148) by Gln298Leu mutant chymosin
191
-
NT/NMN 142-151
-
hydrolysis of the Phe148 variant (cleavage F147-R148) by wild type chymosin
12
-
POm C165-174
-
hydrolysis of the Glu171 variant by D289M/Q298L mutant chymosin
14
-
POm C165-174
-
hydrolysis of the Glu171 variant by Asp289Met mutant chymosin
43
-
POm C165-174
-
hydrolysis of the parent peptide by D289M/Q298L mutant chymosin
52
-
POm C165-174
-
hydrolysis of the Glu171 variant by Gln298Leu mutant chymosin
63
-
POm C165-174
-
hydrolysis of the Glu171 variant by wild type chymosin
90
-
POm C165-174
-
hydrolysis of the parent peptide by Gln298Leu mutant chymosin
101
-
POm C165-174
-
hydrolysis of the parent peptide by Asp289Met mutant chymosin
219
-
POm C165-174
-
hydrolysis of the parent peptide by wild type chymosin
11.7
-
undecapeptide analogue to chymosin sensitive region of bovine kappa casein
-
-
-
44.3
-
undecapeptide analogue to chymosin sensitive region of bovine kappa casein
-
-
-
4.3
-
undecapeptide analogue to chymosin sensitive region of camel kappa casein
-
-
-
5.1
-
undecapeptide analogue to chymosin sensitive region of camel kappa casein
-
-
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
790
-
AFPLEFEREL
-
pH 4.0, 37C
302664
7100
-
AFPLEFFREL
-
pH 4.0, 37C
302666
5400
-
AFPLEFIREL
-
pH 4.0, 37C
302665
160
-
AFPLEFKREL
-
pH 4.0, 37C
302663
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2.5
-
-
-
3.7
-
-
isoform chymosin B
4
-
-
-
4.2
-
-
isoform chymosin A
4.5
-
-, Q9N1p5
recombinant chymosin
4.9
-
-
for proteolysis of undecapeptide analogue to chymosin sensitive region of the bovine kappa casein
5
-
-
-
5.1
-
-
for proteolysis of the undecapeptide analogue to chymosin sensitive region of bovine kappa casein
5.5
-
Q5VK60
-
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
1
5
-
acid-denatured hemoglobin, pH 1: about 80% of maximum activity, pH 5: about 30% of maximum activity
2
6
-
-
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
-
-
4.85
-
-
calculated isoelectric point
5.5
8
Q5VK60
sharp decline of activity up to pH 5.8, moderate decline at higher pH with complete loss of activity at pH 8
5.5
-
-
measured isoelectric point
5.5
-
-
using kappa-casein as substrate
5.6
-
-
-
5.8
-
-
using whole casein as substrate
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
50
-
-
30
-
-
assay at
30
-
Q5VK60
milk clotting activity
37
-
-
-
37
-
-, Q9N1p5
-
42
-
-
for proteolysis of the undecapeptide analogue to chymosin sensitive region of bovine kappa casein
47
-
-
for proteolysis of the undecapeptide analogue to chymosin sensitive region of camel kappa casein
50
-
-
-
55
-
Q5VK60
proteolytic activity
56
-
-
for proteolysis of the undecapeptide analogue to chymosin sensitive region of camel kappa casein
additional information
-
Q5VK60
the substrate temperature was found to influence milk clotting activity, optimum substrate temperature is 65C
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
52.5
-
-
20
60
-
20C: about 25% of maximum activity, 60C: about 50% of maximum activity
25
45
-, Q9N1p5
-
30
60
Q5VK60
milk clotting activity declines moderately up to 55C (80% of maximum activity) and than drops to 40% of activity at 60C proteolytic activity at 30C and 60C approximately 80% of maximum activity
additional information
-
Q5VK60
the substrate temperature was found to influence milk clotting activity, sharp optimum at 65C with complete loss of activity at 75C, less than 20% of activity at 30C
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.55
-
-
isoelectric focusing
4.6
-
-
recombinant chymosin
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Q5VK60
enzyme extracted from abdomasal tissue of 15 days old kid
Manually annotated by BRENDA team
-
transgenic sheep milk
Manually annotated by BRENDA team
Q5VK60
produced in the fourth stomach of milk fed ruminants as inactive precursor prochymosin
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
additional information
-
rennet from abomasal tissue from a local indigenous breed
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30700
-
-
amino acid composition
33000
-
-
gel filtration
35600
-
-
calculated peptide mass, SDS-PAGE
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
-
-
purified chymosin, gel filtration
36300
-
-
two-dimensional electrophoresis technique
36500
-
-
sedimentation, diffusion
40000
-
-
apparent molecular weight, determined by comparison to a Novex Mark12TM
additional information
-
-
-
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 44000, SDS-PAGE
?
Q5VK60
x * 36000, SDS-PAGE
?
-, Q9N1p5
x * 36000, SDS-PAGE, recombinant chymosin
additional information
-
? * 36000, SDS-PAGE
additional information
-
? * 36000, SDS-PAGE, similar enzyme
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
12% glycosylated, oligomannoside attached covalently
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 ACCESSION NO.
LITERATURE
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
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2
6
-
under incubation at 37C 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
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
2
-
-
-
3.5
-
-
rapid loss of activity
5.3
6.3
-
highest stability
7
-
-
rapid loss of activity above
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
-
-
very stable below, suspension of crystals
25
50
-
transgenic chymosin loses activity slightly faster than abomasal enzyme and recombinant enzyme
25
-
-
most stable at pH 5.0, denaturation can be fitted to the two-state irreversible model
40
55
-
no residual clotting activities at temperatures higher than 55C
45
-
-
about 25% loss of activity after 60 min
50
60
-
chymosin is stable up to 50C and a relative milk-clotting activity of 50% is recorded when the temperature is raised to 60C
50
-
-
about 80% loss of activity after 30 min
55
-
-
95% loss of activity after 10 min
55
-
Q5VK60
stable up to 55C, more thermostable than cattle chymosin and equally stable as buffalo chymosin
65
-
-
transition midpoint
additional information
-
-
the enzyme undergoes irreversible, highly scan-rate-dependent thermal denaturation under all the experimental conditions investigated (pH 2-12, 20-50C). Between pH 3.0 and 7.0, only one endotherm characterizes the thermal denaturation of the enzyme. Upon reaching pH 7.5, the denaturation is characterized by two endotherms
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
urea
-
chymosin loses 50% of its activity after incubation in 4.6 M urea at 37C for 30 min
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C, at pH 3.0 and 6.2, 3 days, maximum stability
-
very stable below 5C, suspension of crystals
-
-20C, Tris-HCl buffer, pH 8.0, glycerol
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
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
-
purification by partitioning between aqueous two-phase systems
-
by affinity chromatography MIMO1300 matrix
-
resolves into three major peaks in DEAE chromatography
Q5VK60
similar enzyme
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed as inclusion bodies in Escherichia coli
-
expressed in Escherichia coli and in Saccharomyces cerevisiae
-
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
-
-
-, Q9N1p5
expression in Saccharomyces cerevisiae strain INVSC1
-
expression in Aspergillus niger var. awamori dgr246pyrG
-
prochymosin cDNA, expressed as a NusA fusin protein in Escherichia coli, low level of milk clotting activity after activation at acidic pH
-
expression in Pichia pastoris
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
A115T
-
reduction of Km compared to wild-type
D304A
-
pH optima shift
D304A/T218A
-
double mutant, substrate-specificity-pocket variant
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
D289M/Q298L
-
decreased hydrolytic activity
additional information
-
Asp289Met mutant, Gln298Leu mutant, hydrolytic activity toward peptides having Lys at P'1 decreases significantly, mainly because of a decrease in the turnover values
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
oxidative refolding of solubilized enzyme obtained from inclusion bodies at pH 2 to obtain autoconversion of prochymosin to active chymosin
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
food industry
-
used for the production of dairy products
food industry
-
coagulant for cheese making
food industry
Q5VK60
used as milk coagulant in cheese preparation
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
additional information
-
under typical cheese-making conditions (pH 6.6, 0-2 mM CaCl2) the clotting activity of camel chymosin is ca. 80% higher than the activity of bovine chymosin (average clotting activity of camel chymosin is 70% higher than the activity of bovine chymosin), camel chymosin is more thermostable than bovine chymosin