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
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
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
endopeptidase; peptides, endopeptidase
-
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
chymase
-
-
-
-
chymosin A
-
allelic form
chymosin B
-
-
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
-
prochymosin
inactive precursor
rennin
-
-
-
-
EC 3.4.4.3
-
-
formerly
-
additional information
acid protease, major component of rennet (milk clotting enzyme)
CAS REGISTRY NUMBER
COMMENTARY
9001-98-3
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
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
variant rhizopodiformis
-
-
Manually annotated by BRENDA team
Rhizopus microsporus F518
variant rhizopodiformis
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
-
chymosin, an aspartic protease, is the main enzymatic component of calf rennet
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
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
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
-
?
alpha-casein + H2O
?
show the reaction diagram
-
-
-
-
?
alpha-casein + H2O
?
show the reaction diagram
-
alpha-casein is hydrolyzed extensively
-
-
?
alpha-casein + H2O
?
show the reaction diagram
-
at 50% casein degradation, 15% more of alphaS1-casein with eight phosphate groups is hydrolysed compared with alphaS1-casein with nine phosphate groups in chymosin-induced milk gels
-
-
?
alpha-casein + H2O
?
show the reaction diagram
Rhizopus microsporus F518
-
alpha-casein is hydrolyzed extensively
-
-
?
alphaS-casein + H2O
?
show the reaction diagram
-
-
-
-
?
alphas1-casein + H2O
?
show the reaction diagram
-
-
-
-
?
alphas1-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
-
-
-
-
?
beta-casein + H2O
?
show the reaction diagram
-
occurs during the long period of renneting
-
-
?
beta-casein + H2O
?
show the reaction diagram
-
in sodium caseinate solutions, more than 10% more beta-casein A2 is degraded compared with beta-casein A1 and B at 50% casein degradation
-
-
?
beta-casein + H2O
?
show the reaction diagram
Rhizopus microsporus F518
-
-
-
-
?
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 + 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
-
-
-
-
?
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
-
-
-
?
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
-
?
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
-
?
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
-
-
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
-
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
-
-
-
?
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
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
?
show the reaction diagram
-
the enzyme cleaves the Phe105-Met106 bond
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
the enzyme cleaves the Phe105-Met106 bond of kappa-casein, releasing its predominantly negatively charged C-terminus
-
-
?
kappa-casein + H2O
?
show the reaction diagram
Rhizopus microsporus F518
-
-
-
-
?
kappa-casein + H2O
casein macropeptide + para-kappa-casein
show the reaction diagram
-
-
-
?
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
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
-
?
kappa-casein + H2O
para-kappa-casein + glycopeptide
show the reaction diagram
cleavage between Phe105-Met106
-
?
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
-
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
-
-
-
?
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
-
?
skim milk + H2O
?
show the reaction diagram
-
-
-
-
?
skim milk + H2O
?
show the reaction diagram
-
-
-
-
?
skim milk + H2O
?
show the reaction diagram
-
-
-
-
?
skim milk + H2O
?
show the reaction diagram
Rhizopus microsporus F518
-
-
-
-
?
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 + 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
?
Lys-Pro-Leu-Glu-Phe-Phe(NO2)-Arg-Leu + H2O
Lys-Pro-Leu-Glu-Phe + Phe(NO2)-Arg-Leu
show the reaction diagram
-
-
-
?
additional information
?
-
-
does not cleave alpha-casein and beta-casein
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
alpha-casein + H2O
?
show the reaction diagram
Rhizopus microsporus, Rhizopus microsporus F518
-
alpha-casein is hydrolyzed extensively
-
-
?
alphas1-casein + H2O
?
show the reaction diagram
-
-
-
-
?
alphas1-casein + H2O
?
show the reaction diagram
-
-
-
-
?
beta-casein + H2O
?
show the reaction diagram
Rhizopus microsporus, Rhizopus microsporus F518
-
-
-
-
?
bovine kappa-casein + H2O
para-kappa-casein + caseinomacropeptide
show the reaction diagram
-
the enzyme cleaves the Phe105-Met106 bond of kappa-casein
-
?
kappa-casein + H2O
?
show the reaction diagram
-
-
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
-
-
-
?
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
?
show the reaction diagram
-
the enzyme cleaves the Phe105-Met106 bond
-
-
?
kappa-casein + H2O
?
show the reaction diagram
-
the enzyme cleaves the Phe105-Met106 bond of kappa-casein, releasing its predominantly negatively charged C-terminus
-
-
?
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
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 + glycopeptide
show the reaction diagram
Q5VK60
cleavage between Phe105-Met106
-
?
skim milk + H2O
?
show the reaction diagram
-
-
-
-
?
kappa-casein + H2O
?
show the reaction diagram
Rhizopus microsporus F518
-
-
-
-
?
additional information
?
-
-
does not cleave alpha-casein and beta-casein
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
-
stimulation of milk clotting activity
Ca2+
-
1 mM increases the milk clotting activity by 21.8%
Ca2+
-
maximum milk clotting activity is detected with 20-40 mM Ca2+
Cr3+
-
1 mM increases the milk clotting activity by 15.1%
Zn2+
-
1 mM increases the milk clotting activity by 12%
Mg2+
-
1 mM increases the milk clotting activity by 20.1%
additional information
-
Cu2+, Mn2+, and Co2+ ions have no effect on the enzyme activity
INHIBITORS
ORGANISM
UNIPROT
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
-
Ca2+
-
milkclotting activity decrease when the concentration of Ca2+ is higher than 40 mM
Diazoacetylnorleucine methyl ester
-
in presence of Cu2+
EDTA
-
10 mM EDTA causes inhibition of 73.8% of the milk-clotting activity
Pepstatin
-
-
Pepstatin
-
inhibits recombinant and transgenic chymosin less efficiently than the native enzyme
pepstatin A
-
-
pepstatin A
-
complete inhibition at 0.02 mM
Sn2+
-
28.8% inhibition at 1 mM
iodoacetamide
-
4 mM iodoacetamide causes inhibition of 7.2% of the milk-clotting activity
additional information
-
phenylmethylsulfonyl fluoride does not affect the enzyme activity
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
His-Pro-His-Pro-His-NH2
-
stimulation, part of kappa-casein
additional information
-
when subjected to a low pH, recombinant prochymosin is converted into mature and active chymosin
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
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
-
-
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]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.12
Abz-A-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
Bos taurus
-
hydrolysis by recombinant chymosin
0.16
Abz-A-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
Bos taurus
-
hydrolysis by transgenic chymosin
1.6
Abz-A-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
Bos taurus
-
hydrolysis by calf chymosin
1.4
Abz-A-A-F-F-A-A-p-nitroanilide
Bos taurus
-
hydrolysis by calf chymosin
0.02
Abz-A-A-F-F-A-N-(2,4-dinitrophenyl)-ethylenediamine
Bos taurus
-
hydrolysis by transgenic chymosin
0.11
Abz-A-A-F-F-A-N-(2,4-dinitrophenyl)-ethylenediamine
Bos taurus
-
hydrolysis by recombinant chymosin
0.8
Abz-A-A-F-F-A-N-(2,4-dinitrophenyl)-ethylenediamine
Bos taurus
-
hydrolysis by calf chymosin
0.27
Abz-A-A-F-F-N-(2,4-dinitrophenyl)-ethylenediamine
Bos taurus
-
hydrolysis by calf chymosin
0.48
Abz-A-A-F-F-p-nitroanilide
Bos taurus
-
hydrolysis by calf chymosin
0.06
Abz-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
Bos taurus
-
hydrolysis by transgenic chymosin or recombinat chymosin
0.14
Abz-A-F-F-A-A-N-(2,4-dinitrophenyl)-ethylenediamine
Bos taurus
-
hydrolysis by calf chymosin
2600
AFPLEFEREL
Bos taurus
-
pH 4.0, 37C
290
AFPLEFFREL
Bos taurus
-
pH 4.0, 37C
427
AFPLEFIREL
Bos taurus
-
pH 4.0, 37C
29
AFPLEFKREL
Bos taurus
-
pH 4.0, 37C
2 - 8
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the Phe148 variant (cleavage L147-F148) by wild type chymosin
12
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the Phe148 variant (cleavage L147-F148) by Gln298Leu mutant chymosin
16
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the Phe148 variant (cleavage L147-F148) by D289M/Q298L mutant chymosin
17
NT/NMN 142-151
Callithrix jacchus
-
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
Callithrix jacchus
-
hydrolysis of the Glu148 variant by D289M/Q298L mutant chymosin
27
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the parent peptide by D289M/Q298L mutant chymosin
30
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the parent peptide by Gln298Leu mutant chymosin
32
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the Glu148 variant by Asp289Met mutant chymosin
35
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the parent peptide by Asp289Met mutant chymosin
39
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the Phe148 variant (cleavage L147-F148) by Asp289Met mutant chymosin
41
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the parent peptide by wild type chymosin
42
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the Phe148 variant (cleavage F147-R148) by Asp289Met mutant chymosin
46
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the Glu148 variant by wild type chymosin
49
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the Phe148 variant (cleavage F147-R148) by Gln298Leu mutant chymosin
191
NT/NMN 142-151
Callithrix jacchus
-
hydrolysis of the Phe148 variant (cleavage F147-R148) by wild type chymosin
12
POm C165-174
Callithrix jacchus
-
hydrolysis of the Glu171 variant by D289M/Q298L mutant chymosin
14
POm C165-174
Callithrix jacchus
-
hydrolysis of the Glu171 variant by Asp289Met mutant chymosin
43
POm C165-174
Callithrix jacchus
-
hydrolysis of the parent peptide by D289M/Q298L mutant chymosin
52
POm C165-174
Callithrix jacchus
-
hydrolysis of the Glu171 variant by Gln298Leu mutant chymosin
63
POm C165-174
Callithrix jacchus
-
hydrolysis of the Glu171 variant by wild type chymosin
90
POm C165-174
Callithrix jacchus
-
hydrolysis of the parent peptide by Gln298Leu mutant chymosin
101
POm C165-174
Callithrix jacchus
-
hydrolysis of the parent peptide by Asp289Met mutant chymosin
219
POm C165-174
Callithrix jacchus
-
hydrolysis of the parent peptide by wild type chymosin
11.7
undecapeptide analogue to chymosin sensitive region of bovine kappa casein
Camelus dromedarius
-
-
-
44.3
undecapeptide analogue to chymosin sensitive region of bovine kappa casein
Bos taurus
-
-
-
4.3
undecapeptide analogue to chymosin sensitive region of camel kappa casein
Bos taurus
-
-
-
5.1
undecapeptide analogue to chymosin sensitive region of camel kappa casein
Camelus dromedarius
-
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
790
AFPLEFEREL
Bos taurus
-
pH 4.0, 37C
12112
7100
AFPLEFFREL
Bos taurus
-
pH 4.0, 37C
12114
5400
AFPLEFIREL
Bos taurus
-
pH 4.0, 37C
12113
160
AFPLEFKREL
Bos taurus
-
pH 4.0, 37C
40659
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
1.987
-
crude enzyme, in 50 mM acetate buffer (pH 5.6) at 37C
7.272
-
after 33.2fold purification, in 50 mM acetate buffer (pH 5.6) at 37C
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
2.5
-
-
3.7
-
isoform chymosin B
4.2
-
isoform chymosin A
4.5
recombinant chymosin
4.9
-
for proteolysis of undecapeptide analogue to chymosin sensitive region of the bovine kappa casein
5
-
optimum for milk clotting activity
5.1
-
for proteolysis of the undecapeptide analogue to chymosin sensitive region of bovine kappa casein
5.5
-
-
pH RANGE
ORGANISM
UNIPROT
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
5.1 - 6.7
-
at pH 5.1, the relative clotting activity decreases to 50% and further declines to 17% and 10% at pH 5.6 and pH 6.7, respectively
5.5
-
measured isoelectric point
5.5
-
using kappa-casein as substrate
5.5 - 8
sharp decline of activity up to pH 5.8, moderate decline at higher pH with complete loss of activity at pH 8
5.6
-
-
5.8
-
using whole casein as substrate
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
-
assay at
30
milk clotting activity
30 - 50
-
-
37
-
-
42
-
for proteolysis of the undecapeptide analogue to chymosin sensitive region of bovine kappa casein
45 - 55
-
optimum for milk clotting activity
47
-
for proteolysis of the undecapeptide analogue to chymosin sensitive region of camel kappa casein
50
-
-
55
proteolytic activity
56
-
for proteolysis of the undecapeptide analogue to chymosin sensitive region of camel kappa casein
additional information
the substrate temperature was found to influence milk clotting activity, optimum substrate temperature is 65C
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 52.5
-
-
20 - 60
-
20C: about 25% of maximum activity, 60C: about 50% of maximum activity
30 - 60
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
35 - 60
-
relative activity of 50% is observed at temperatures of 35C and 60C. Chymosin activity completely ceases below 20C or above 70C
additional information
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
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.55
-
isoelectric focusing
4.6
-
recombinant chymosin
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
enzyme extracted from abdomasal tissue of 15 days old kid
Manually annotated by BRENDA team
-
transgenic sheep milk
Manually annotated by BRENDA team
produced in the fourth stomach of milk fed ruminants as inactive precursor prochymosin
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
additional information
-
rennet from abomasal tissue from a local indigenous breed
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30700
-
amino acid composition
36882
33000
-
gel filtration
36882
35600
-
calculated peptide mass, SDS-PAGE
667435
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
708236
36000
-
purified chymosin, gel filtration
669642
36300
-
two-dimensional electrophoresis technique
36872
36500
-
sedimentation, diffusion
36882
40000
-
apparent molecular weight, determined by comparison to a Novex Mark12TM
667435
additional information
-
-
36882
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 44000, SDS-PAGE
?
x * 36000, SDS-PAGE
?
x * 36000, SDS-PAGE, recombinant chymosin
?
-
x * about 40000, mass spectrometry
?
-
x * 37000, SDS-PAGE
?
x * 42000, recombinant enzyme, SDS-PAGE
?
-
x * 40000, SDS-PAGE
?
-
x * 40000, active chymosin, SDS-PAGE; x * 42000, deglycosylated recombinant enzyme, SDS-PAGE; x * 45000, glycosylated recombinant enzyme, SDS-PAGE
?
Rhizopus microsporus F518
-
x * 37000, SDS-PAGE
-
additional information
-
? * 36000, SDS-PAGE
additional information
-
? * 36000, SDS-PAGE, similar enzyme
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
glycoprotein
-
glycosylation at Asn291 significantly decreases enzyme activity
glycoprotein
-
glycosylation at Asn291 significantly decreases enzyme activity
glycoprotein
-
doubly glycosylated camel chymosin has slightly higher catalytic efficiency
glycoprotein
-
-
glycoprotein
-
12% glycosylated, oligomannoside attached covalently
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
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
-
unglycosylated chymosin, vapor diffusion method, using 100 mM NaH2PO4 pH 5.5, 1.5 M NaCl
-
doubly and singly glycosylated variants of chymosin, vapor diffusion method, using 2 M ammonium sulfate, 100 mM bis-Tris buffer in the pH range 5.1-6.5
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
2
-
-
36882
2 - 3.8
-
-
667435
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
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
2 - 8
-
the enzyme retains 80% of activity within pH 2.0-8.0
731169
2.5 - 6.5
-
the recombinant enzyme is highly active and stable over a wide pH range (from 2.5 to 6.0) at 20C for 8 h. Relative clotting activity declines when the recombinant chymosin is kept at a pH value above 6.0. No detectable clotting activity at pH 6.5
732866
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
LITERATURE
0 - 50
-
after 8 h of incubation, between 0 and 20C the enzyme shows 100% activity, while at 30C and 40C 70% and 35% is retained, respectively. The enzyme is inactive after 8 h at 50C
732866
5
-
very stable below, suspension of crystals
36882
25
-
most stable at pH 5.0, denaturation can be fitted to the two-state irreversible model
650661
25 - 50
-
transgenic chymosin loses activity slightly faster than abomasal enzyme and recombinant enzyme
667763
40
-
the enzyme is stable until 40C for 30 min while losing 40 and 80% of its activity after incubation at 45 and 50C for 30 min, respectively
731169
40 - 55
-
no residual clotting activities at temperatures higher than 55C
667435
45
-
about 25% loss of activity after 60 min
36874
50
-
about 80% loss of activity after 30 min
36874
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
708236
55
-
95% loss of activity after 10 min
36874
55
stable up to 55C, more thermostable than cattle chymosin and equally stable as buffalo chymosin
686199
56
-
chymosin-mediated hydrolysis of alphas1 casein is slower in cheeses treated at 56C
732234
56
-
chymosin-mediated hydrolysis of alphas1 casein is slower in cheeses treated at 56C
732235
60
-
melting temperature of the singly glycosylated enzyme
731064
65
-
transition midpoint
650661
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
650661
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
for prochymosins activation, the supernatant is adjusted to pH 2.0 and incubated for 2 h at 25C, then adjusted to pH 6.3 and left again at 25C for 1 h
ORGANIC SOLVENT
ORGANISM
UNIPROT
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
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
-
0-20C, at pH 5.5, 8 h, no loss of activity
-
40C, at pH 5.5, 8 h, 37.5% residual activity
-
50C, at pH 5.5, complete loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
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
-
phenyl Superose column chromatography
-
purification by partitioning between aqueous two-phase systems
-
ultrafiltration and Superdex 75 gel filtration
-
by affinity chromatography MIMO1300 matrix
-
phenyl Superose column chromatography
-
metal affinity chromatography
resolves into three major peaks in DEAE chromatography
similar enzyme
-
ammonium sulfate precipitation, Q Sepharose column chromatography, and Sephadex S-100 gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed as inclusion bodies in Escherichia coli
-
expressed in Escherichia coli and in Saccharomyces cerevisiae
-
expressed in Pichia pastoris strain GS115
-
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
-
expression in Saccharomyces cerevisiae strain INVSC1
-
expressed in Pichia pastoris strain GS115
-
expression in Aspergillus niger var. awamori dgr246pyrG
-
expressed in Escherichia coli BL21(DE3) cells
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
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
additional information
-
the loss of the first three residues of camel chymosin significantly decreases enzyme activity
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
-
using 8 M urea, at 25C
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
food industry
-
used for the production of dairy products
food industry
-
coagulant for cheese making
food industry
-
the enzyme is used industrially in cheese production
food industry
-
chymosin constitutes a traditional ingredient for enzymatic milk coagulation in cheese making
food industry
-
the enzyme is used for the production of Reggianito cooked cheese
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
food industry
-
the enzyme is used industrially in cheese production
food industry
-
chymosin constitutes a traditional ingredient for enzymatic milk coagulation in cheese making
food industry
used as milk coagulant in cheese preparation
food industry
the enzyme is used as milk coagulant in the cheese industry
food industry
-
the enzyme is used for the production of Reggianito cooked cheese
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