CHYM_CAMDR
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 03-AUG-2022, entry version 102.
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 kappa-
CC 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 kappa-
CC 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 kappa-
CC 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 kappa-
CC chain of casein (double-glycosylated 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 kappa-
CC chain of casein (double-glycosylated form of chymosin at pH 6.0)
CC {ECO:0000269|PubMed:25726113};
CC KM=8 uM for a labeled peptide substrate of 98-112 of bovine kappa-
CC chain of casein (double-glycosylated form of chymosin at pH 5.5)
CC {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 kappa-
CC chain of casein (unglycosylated form of chymosin at pH 6.7), kcat is
CC 53 sec(-1) for a labeled peptide substrate of 98-112 of bovine kappa-
CC chain of casein (unglycosylated form of chymosin at pH 6.0), kcat is
CC 47 sec(-1) for a labeled peptide substrate of 98-112 of bovine kappa-
CC chain of casein (unglycosylated form of chymosin at pH 5.5), kcat is
CC 14 sec(-1) for a labeled peptide substrate of 98-112 of bovine kappa-
CC chain of casein (double-glycosylated form of chymosin at pH 6.7),
CC kcat is 65 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 6.0) and kcat is 59 sec(-1) for a labeled peptide substrate of 98-
CC 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 iPTMnet; Q9GK11; -.
DR Ensembl; ENSCDRT00005002138; ENSCDRP00005001942; ENSCDRG00005001417.
DR GeneID; 105085668; -.
DR KEGG; cdk:105085668; -.
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; 2.40.70.10; -; 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; PTHR47966; 1.
DR Pfam; PF07966; A1_Propeptide; 1.
DR Pfam; PF00026; Asp; 1.
DR PRINTS; PR00792; PEPSIN.
DR SUPFAM; SSF50630; SSF50630; 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
