ID   CHYM_CAMDR              Reviewed;         381 AA.
AC   Q9GK11;
DT   30-NOV-2016, integrated into UniProtKB/Swiss-Prot.
DT   01-MAR-2001, sequence version 1.
DT   22-FEB-2023, entry version 104.
DE   RecName: Full=Chymosin {ECO:0000303|PubMed:16488399, ECO:0000312|EMBL:CAC19554.1};
DE            EC=3.4.23.4 {ECO:0000269|PubMed:16488399, ECO:0000269|PubMed:23633601, ECO:0000269|PubMed:25726113, ECO:0000269|PubMed:25837439};
DE   Flags: Precursor;
GN   Name=CYM {ECO:0000305};
OS   Camelus dromedarius (Dromedary) (Arabian camel).
OC   Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC   Eutheria; Laurasiatheria; Artiodactyla; Tylopoda; Camelidae; Camelus.
OX   NCBI_TaxID=9838 {ECO:0000312|EMBL:CAC19554.1};
RN   [1] {ECO:0000312|EMBL:CAC19554.1}
RP   NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY,
RP   BIOPHYSICOCHEMICAL PROPERTIES, AND BIOTECHNOLOGY.
RC   TISSUE=Gastric mucosa {ECO:0000303|PubMed:16488399,
RC   ECO:0000312|EMBL:CAC19554.1};
RX   PubMed=16488399; DOI=10.1016/j.bbrc.2006.02.014;
RA   Kappeler S.R., van den Brink H.J.M., Rahbek-Nielsen H., Farah Z., Puhan Z.,
RA   Hansen E.B., Johansen E.;
RT   "Characterization of recombinant camel chymosin reveals superior properties
RT   for the coagulation of bovine and camel milk.";
RL   Biochem. Biophys. Res. Commun. 342:647-654(2006).
RN   [2] {ECO:0007744|PDB:4AA9}
RP   PROTEIN SEQUENCE OF 59-74, X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF
RP   62-381, FUNCTION, CATALYTIC ACTIVITY, IDENTIFICATION BY MASS SPECTROMETRY,
RP   ACTIVE SITE, GLYCOSYLATION AT ASN-158, AND DISULFIDE BONDS.
RX   PubMed=23633601; DOI=10.1107/s0907444913003260;
RA   Langholm Jensen J., Molgaard A., Navarro Poulsen J.C., Harboe M.K.,
RA   Simonsen J.B., Lorentzen A.M., Hjerno K., van den Brink J.M., Qvist K.B.,
RA   Larsen S.;
RT   "Camel and bovine chymosin: the relationship between their structures and
RT   cheese-making properties.";
RL   Acta Crystallogr. D 69:901-913(2013).
RN   [3]
RP   FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX   PubMed=25726113; DOI=10.3168/jds.2014-8672;
RA   Jensen J.L., Jacobsen J., Moss M.L., Rasmussen F., Qvist K.B., Larsen S.,
RA   van den Brink J.M.;
RT   "The function of the milk-clotting enzymes bovine and camel chymosin
RT   studied by a fluorescence resonance energy transfer assay.";
RL   J. Dairy Sci. 98:2853-2860(2015).
RN   [4]
RP   FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP   BIOTECHNOLOGY.
RX   PubMed=25837439; DOI=10.1016/j.pep.2015.03.012;
RA   Wang N., Wang K.Y., Li G., Guo W., Liu D.;
RT   "Expression and characterization of camel chymosin in Pichia pastoris.";
RL   Protein Expr. Purif. 111:75-81(2015).
CC   -!- FUNCTION: Chymosin is synthesized in the mucosa of the abomasum (fourth
CC       stomach) of young (unweaned) ruminants (PubMed:16488399). The enzyme
CC       hydrolyzes casein to paracasein (PubMed:16488399, PubMed:23633601,
CC       PubMed:25726113, PubMed:25837439). {ECO:0000269|PubMed:16488399,
CC       ECO:0000269|PubMed:23633601, ECO:0000269|PubMed:25726113,
CC       ECO:0000269|PubMed:25837439}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=Broad specificity similar to that of pepsin A. Clots milk by
CC         cleavage of a single 104-Ser-Phe-|-Met-Ala-107 bond in kappa-chain of
CC         casein.; EC=3.4.23.4; Evidence={ECO:0000269|PubMed:16488399,
CC         ECO:0000269|PubMed:23633601, ECO:0000269|PubMed:25726113,
CC         ECO:0000269|PubMed:25837439};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=56 uM for undecapeptide analog to chymosin sensitive region of
CC         camel kappa-chain of casein {ECO:0000269|PubMed:16488399};
CC         KM=77 uM for undecapeptide analog to chymosin sensitive region of
CC         bovine kappa-chain of casein {ECO:0000269|PubMed:16488399};
CC         KM=18 uM for a labeled peptide substrate of 98-112 of bovine
CC         kappa-chain of casein (unglycosylated form of chymosin at pH 6.7)
CC         {ECO:0000269|PubMed:25726113};
CC         KM=11 uM for a labeled peptide substrate of 98-112 of bovine
CC         kappa-chain of casein (unglycosylated form of chymosin at pH 6.0)
CC         {ECO:0000269|PubMed:25726113};
CC         KM=7 uM for a labeled peptide substrate of 98-112 of bovine
CC         kappa-chain of casein (unglycosylated form of chymosin at pH 5.5)
CC         {ECO:0000269|PubMed:25726113};
CC         KM=20 uM for a labeled peptide substrate of 98-112 of bovine
CC         kappa-chain of casein (double-glycosylated form of chymosin at pH
CC         6.7) {ECO:0000269|PubMed:25726113};
CC         KM=11 uM for a labeled peptide substrate of 98-112 of bovine
CC         kappa-chain of casein (double-glycosylated form of chymosin at pH
CC         6.0) {ECO:0000269|PubMed:25726113};
CC         KM=8 uM for a labeled peptide substrate of 98-112 of bovine
CC         kappa-chain of casein (double-glycosylated form of chymosin at pH
CC         5.5) {ECO:0000269|PubMed:25726113};
CC         Note=kcat is 5.1 sec(-1) for undecapeptide analog to chymosin
CC         sensitive region of camel kappa-chain of casein and kcat is 11.7
CC         sec(-1) for undecapeptide analog to chymosin sensitive region of
CC         bovine kappa-chain of casein (PubMed:16488399). Activity is 462
CC         International Milk-Clotting Units (IMCU) per mg enzyme. kcat is 11
CC         sec(-1) for a labeled peptide substrate of 98-112 of bovine
CC         kappa-chain of casein (unglycosylated form of chymosin at pH 6.7),
CC         kcat is 53 sec(-1) for a labeled peptide substrate of 98-112 of
CC         bovine kappa-chain of casein (unglycosylated form of chymosin at pH
CC         6.0), kcat is 47 sec(-1) for a labeled peptide substrate of 98-112 of
CC         bovine kappa-chain of casein (unglycosylated form of chymosin at pH
CC         5.5), kcat is 14 sec(-1) for a labeled peptide substrate of 98-112 of
CC         bovine kappa-chain of casein (double-glycosylated form of chymosin at
CC         pH 6.7), kcat is 65 sec(-1) for a labeled peptide substrate of 98-112
CC         of bovine kappa-chain of casein (double-glycosylated form of chymosin
CC         at pH 6.0) and kcat is 59 sec(-1) for a labeled peptide substrate of
CC         98-112 of bovine kappa-chain of casein (double-glycosylated form of
CC         chymosin at pH 5.5) (PubMed:25726113). Activity is 400 IMCU per ml
CC         enzyme. Supplemental CaCl(2) at concentration between 20-40 mM is
CC         optimal for stable enzyme activity (PubMed:25837439).
CC         {ECO:0000269|PubMed:16488399, ECO:0000269|PubMed:25726113,
CC         ECO:0000269|PubMed:25837439};
CC       pH dependence:
CC         Optimum pH is about 5.1 (PubMed:16488399). Activity decreases with
CC         increasing pH values (PubMed:25837439). {ECO:0000269|PubMed:16488399,
CC         ECO:0000269|PubMed:25837439};
CC       Temperature dependence:
CC         Optimum temperature is between 45-55 degrees Celsius
CC         (PubMed:16488399, PubMed:25837439). 50% activity at 35 and 60 degrees
CC         Celsius. No activity below 20 or above 70 degrees Celsius
CC         (PubMed:25837439). {ECO:0000269|PubMed:16488399,
CC         ECO:0000269|PubMed:25837439};
CC   -!- BIOTECHNOLOGY: The extraordinary high clotting activity, combined with
CC       its very low non-specific activity may be useful in the production of
CC       cheese types, for which a bitter taste is unfavorable, for example
CC       Mascarpone type cheese (PubMed:16488399). Large-scale expression of
CC       camel chymosin gene in P.pastoris could represent an excellent system
CC       for producing active chymosin for potential use in the commercial
CC       production of cheese (PubMed:25837439). {ECO:0000269|PubMed:16488399,
CC       ECO:0000269|PubMed:25837439}.
CC   -!- SIMILARITY: Belongs to the peptidase A1 family. {ECO:0000255,
CC       ECO:0000255|PROSITE-ProRule:PRU01103, ECO:0000255|RuleBase:RU000454,
CC       ECO:0000305}.
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DR   EMBL; AJ131677; CAC19554.1; -; mRNA.
DR   RefSeq; NP_001290503.1; NM_001303574.1.
DR   PDB; 4AA9; X-ray; 1.60 A; A=62-381.
DR   PDBsum; 4AA9; -.
DR   AlphaFoldDB; Q9GK11; -.
DR   SMR; Q9GK11; -.
DR   STRING; 9838.ENSCDRP00005001942; -.
DR   MEROPS; A01.006; -.
DR   GlyCosmos; Q9GK11; 2 sites, No reported glycans.
DR   iPTMnet; Q9GK11; -.
DR   Ensembl; ENSCDRT00005002138; ENSCDRP00005001942; ENSCDRG00005001417.
DR   GeneID; 105085668; -.
DR   KEGG; cdk:105085668; -.
DR   OrthoDB; 1120702at2759; -.
DR   BRENDA; 3.4.23.4; 1085.
DR   GO; GO:0004190; F:aspartic-type endopeptidase activity; IDA:UniProtKB.
DR   GO; GO:0007586; P:digestion; IEA:UniProtKB-KW.
DR   GO; GO:0051603; P:proteolysis involved in protein catabolic process; IDA:UniProtKB.
DR   CDD; cd05478; pepsin_A; 1.
DR   Gene3D; 6.10.140.60; -; 1.
DR   Gene3D; 2.40.70.10; Acid Proteases; 2.
DR   InterPro; IPR001461; Aspartic_peptidase_A1.
DR   InterPro; IPR001969; Aspartic_peptidase_AS.
DR   InterPro; IPR012848; Aspartic_peptidase_N.
DR   InterPro; IPR034162; Pepsin_A.
DR   InterPro; IPR033121; PEPTIDASE_A1.
DR   InterPro; IPR021109; Peptidase_aspartic_dom_sf.
DR   PANTHER; PTHR47966; BETA-SITE APP-CLEAVING ENZYME, ISOFORM A-RELATED; 1.
DR   PANTHER; PTHR47966:SF13; CHYMOSIN; 1.
DR   Pfam; PF07966; A1_Propeptide; 1.
DR   Pfam; PF00026; Asp; 1.
DR   PRINTS; PR00792; PEPSIN.
DR   SUPFAM; SSF50630; Acid proteases; 1.
DR   PROSITE; PS00141; ASP_PROTEASE; 2.
DR   PROSITE; PS51767; PEPTIDASE_A1; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Aspartyl protease; Digestion; Direct protein sequencing;
KW   Disulfide bond; Glycoprotein; Hydrolase; Protease; Signal; Zymogen.
FT   SIGNAL          1..16
FT                   /evidence="ECO:0000250|UniProtKB:Q9N2D2"
FT   PROPEP          17..58
FT                   /note="Activation peptide"
FT                   /evidence="ECO:0000305|PubMed:23633601"
FT                   /id="PRO_0000438462"
FT   CHAIN           59..381
FT                   /note="Chymosin"
FT                   /id="PRO_5004326438"
FT   DOMAIN          74..378
FT                   /note="Peptidase A1"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU01103"
FT   ACT_SITE        92
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU01103,
FT                   ECO:0000269|PubMed:23633601"
FT   ACT_SITE        274
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU01103,
FT                   ECO:0000269|PubMed:23633601"
FT   CARBOHYD        158
FT                   /note="N-linked (GlcNAc...) asparagine"
FT                   /evidence="ECO:0000269|PubMed:23633601,
FT                   ECO:0007744|PDB:4AA9"
FT   CARBOHYD        349
FT                   /note="N-linked (GlcNAc...) asparagine"
FT                   /evidence="ECO:0000305|PubMed:23633601"
FT   DISULFID        105..110
FT                   /evidence="ECO:0000269|PubMed:23633601,
FT                   ECO:0007744|PDB:4AA9"
FT   DISULFID        265..269
FT                   /evidence="ECO:0000269|PubMed:23633601,
FT                   ECO:0007744|PDB:4AA9"
FT   DISULFID        308..341
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU01103,
FT                   ECO:0000269|PubMed:23633601, ECO:0007744|PDB:4AA9"
FT   STRAND          75..80
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   TURN            81..84
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          85..92
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          98..102
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   HELIX           108..111
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   HELIX           118..120
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          125..135
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          138..151
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          154..166
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   HELIX           170..173
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          178..182
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   HELIX           186..188
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   HELIX           196..202
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          206..214
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          217..220
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          223..227
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   HELIX           231..233
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          234..242
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   TURN            246..249
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          250..253
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          255..258
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          261..265
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          269..273
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          278..283
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   HELIX           284..293
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          304..306
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   HELIX           308..313
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          317..321
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          324..328
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   HELIX           330..333
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          334..337
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          340..348
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          354..356
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   HELIX           358..361
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          364..369
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   TURN            370..373
FT                   /evidence="ECO:0007829|PDB:4AA9"
FT   STRAND          374..380
FT                   /evidence="ECO:0007829|PDB:4AA9"
SQ   SEQUENCE   381 AA;  42083 MW;  24BADB57B2E7FDD7 CRC64;
     MRCLVVLLAA LALSQASGIT RIPLHKGKTL RKALKERGLL EDFLQRQQYA VSSKYSSLGK
     VAREPLTSYL DSQYFGKIYI GTPPQEFTVV FDTGSSDLWV PSIYCKSNVC KNHHRFDPRK
     SSTFRNLGKP LSIHYGTGSM EGFLGYDTVT VSNIVDPNQT VGLSTEQPGE VFTYSEFDGI
     LGLAYPSLAS EYSVPVFDNM MDRHLVARDL FSVYMDRNGQ GSMLTLGAID PSYYTGSLHW
     VPVTLQQYWQ FTVDSVTING VAVACVGGCQ AILDTGTSVL FGPSSDILKI QMAIGATENR
     YGEFDVNCGN LRSMPTVVFE INGRDYPLSP SAYTSKDQGF CTSGFQGDNN SELWILGDVF
     IREYYSVFDR ANNRVGLAKA I
//