ID   SUBC_BACLI              Reviewed;         379 AA.
AC   P00780; Q9F943;
DT   21-JUL-1986, integrated into UniProtKB/Swiss-Prot.
DT   18-SEP-2019, sequence version 2.
DT   02-SEP-2026, entry version 178.
DE   RecName: Full=Subtilisin Carlsberg {ECO:0000303|PubMed:4967581};
DE            EC=3.4.21.62 {ECO:0000269|PubMed:11109488, ECO:0000269|Ref.4};
DE   Flags: Precursor;
GN   Name=subC {ECO:0000312|EMBL:CAB56500.1}; Synonyms=apr;
OS   Bacillus licheniformis.
OC   Bacteria; Bacillati; Bacillota; Bacilli; Caryophanales; Bacillaceae;
OC   Bacillus.
OX   NCBI_TaxID=1402;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=NCIMB 6816 / CCM 2182 / NCDO 727 / NCTC 6816;
RX   PubMed=3001653; DOI=10.1093/nar/13.24.8913;
RA   Jacobs M., Eliasson M., Uhlen M., Flock J.-I.;
RT   "Cloning, sequencing and expression of subtilisin Carlsberg from Bacillus
RT   licheniformis.";
RL   Nucleic Acids Res. 13:8913-8926(1985).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-374, FUNCTION, CATALYTIC ACTIVITY,
RP   BIOPHYSICOCHEMICAL PROPERTIES, AND SUBCELLULAR LOCATION.
RC   STRAIN=NCIMB 6816 / CCM 2182 / NCDO 727 / NCTC 6816;
RX   PubMed=11109488; DOI=10.1139/w00-085;
RA   Evans K.L., Crowder J., Miller E.S.;
RT   "Subtilisins of Bacillus spp. hydrolyze keratin and allow growth on
RT   feathers.";
RL   Can. J. Microbiol. 46:1004-1011(2000).
RN   [3]
RP   PROTEIN SEQUENCE OF 106-379.
RX   PubMed=4967581; DOI=10.1016/s0021-9258(18)93461-7;
RA   Smith E.L., Delange R.J., Evans W.H., Landon M., Markland F.S.;
RT   "Subtilisin Carlsberg. V. The complete sequence; comparison with subtilisin
RT   BPN'; evolutionary relationships.";
RL   J. Biol. Chem. 243:2184-2191(1968).
RN   [4]
RP   FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX   DOI=10.1007/BF01569566;
RA   Peters K., Pauli D., Hache H., Boteva R.N., Genov N.C., Fittkau S.;
RT   "Subtilisin DY-Kinetic characterization and comparison with related
RT   proteinases.";
RL   Curr. Microbiol. 18:171-177(1989).
RN   [5] {ECO:0007744|PDB:1SEL}
RP   X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) OF 106-379 OF MUTANT
RP   SELENOCYSTEINE-325 IN COMPLEX WITH CALCIUM, AND COFACTOR.
RX   PubMed=8512925; DOI=10.1021/bi00075a007;
RA   Syed R., Wu Z.P., Hogle J.M., Hilvert D.;
RT   "Crystal structure of selenosubtilisin at 2.0-A resolution.";
RL   Biochemistry 32:6157-6164(1993).
RN   [6] {ECO:0007744|PDB:1AV7, ECO:0007744|PDB:1AVT, ECO:0007744|PDB:1VSB, ECO:0007744|PDB:3VSB}
RP   X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) OF 106-379, AND ACTIVITY REGULATION.
RX   PubMed=9425066; DOI=10.1021/bi971166o;
RA   Stoll V.S., Eger B.T., Hynes R.C., Martichonok V., Jones J.B., Pai E.F.;
RT   "Differences in binding modes of enantiomers of 1-acetamido boronic acid
RT   based protease inhibitors: crystal structures of gamma-chymotrypsin and
RT   subtilisin Carlsberg complexes.";
RL   Biochemistry 37:451-462(1998).
CC   -!- FUNCTION: Subtilisin is an extracellular alkaline serine protease, it
CC       catalyzes the hydrolysis of proteins and peptide amides
CC       (PubMed:11109488, Ref.4). Shows high specificity for aromatic and
CC       hydrophobic amino acids in the P1 substrate position (PubMed:11109488).
CC       May play an important role in the degradation of feather keratin
CC       (PubMed:11109488). {ECO:0000269|PubMed:11109488, ECO:0000269|Ref.4}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=Hydrolysis of proteins with broad specificity for peptide
CC         bonds, and a preference for a large uncharged residue in P1.
CC         Hydrolyzes peptide amides.; EC=3.4.21.62;
CC         Evidence={ECO:0000269|PubMed:11109488, ECO:0000269|Ref.4};
CC   -!- COFACTOR:
CC       Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC         Evidence={ECO:0000269|PubMed:8512925};
CC       Note=Binds 2 calcium ions per subunit. {ECO:0000269|PubMed:8512925};
CC   -!- ACTIVITY REGULATION: Inhibited by p-chlorophenyl and 1-naphthyl boronic
CC       acid derivatives. {ECO:0000269|PubMed:9425066}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=1.2 mM for Suc-Ala(2)-Ala-pNA {ECO:0000269|Ref.4};
CC         KM=0.79 mM for Suc-Ala(2)-Phe-pNA {ECO:0000269|Ref.4};
CC         KM=0.851 mM for Suc-Ala(2)-Phe-pNA {ECO:0000269|PubMed:11109488};
CC         KM=0.22 mM for MeO-Suc-Ala(2)-Phe-pNA {ECO:0000269|Ref.4};
CC         KM=5.6 mM for Suc-Gly(2)-Phe-pNA {ECO:0000269|PubMed:11109488};
CC         KM=0.334 mM for Suc-Ala(2)-Pro-Phe-pNA {ECO:0000269|PubMed:11109488};
CC         KM=0.062 mM for Suc-Ala(2)-Pro-Leu-pNA {ECO:0000269|PubMed:11109488};
CC         KM=0.422 mM for Suc-Ala(2)-Pro-Arg-pNA {ECO:0000269|PubMed:11109488};
CC         KM=0.245 mM for Suc-Ala(2)-Pro-Glu-pNA {ECO:0000269|PubMed:11109488};
CC         KM=0.042 mM for Suc-Ala-Glu-Pro-Phe-pNA
CC         {ECO:0000269|PubMed:11109488};
CC         Note=kcat is 1.6 sec(-1) with Suc-Ala(2)-Ala-pNA as substrate. kcat
CC         is 57 sec(-1) with Suc-Ala(2)-Phe-pNA as substrate. kcat is 97
CC         sec(-1) with MeO-Suc-Ala(2)-Phe-pNA as substrate (Ref.4). kcat is
CC         0.990 sec(-1) with Suc-Gly(2)-Phe-pNA as substrate. kcat is 20.3
CC         sec(-1) with Suc-Ala(2)-Phe-pNA as substrate. kcat is 646 sec(-1)
CC         with Suc-Ala(2)-Pro-Phe-pNA as substrate. kcat is 137 sec(-1) with
CC         Suc-Ala(2)-Pro-Leu-pNA as substrate. kcat is 5.31 sec(-1) with
CC         Suc-Ala(2)-Pro-Arg-pNA as substrate. kcat is 1.92 sec(-1) with
CC         Suc-Ala(2)-Pro-Glu-pNA as substrate. kcat is 375 sec(-1) with
CC         Suc-Ala-Glu-Pro-Phe-pNA as substrate (PubMed:11109488).
CC         {ECO:0000269|PubMed:11109488, ECO:0000269|Ref.4};
CC   -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:11109488}.
CC   -!- BIOTECHNOLOGY: Used as a detergent protease. Sold under the name
CC       Alcalase by Novozymes.
CC   -!- SIMILARITY: Belongs to the peptidase S8 family. {ECO:0000255|PROSITE-
CC       ProRule:PRU01240}.
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DR   EMBL; X03341; CAB56500.1; -; Genomic_DNA.
DR   EMBL; AF205189; AAG31026.1; -; Genomic_DNA.
DR   PIR; A24111; SUBSCL.
DR   PDB; 1AF4; X-ray; 2.60 A; A=106-379.
DR   PDB; 1AV7; X-ray; 2.60 A; A=106-379.
DR   PDB; 1AVT; X-ray; 2.00 A; A=106-379.
DR   PDB; 1BE6; X-ray; 2.15 A; A=106-379.
DR   PDB; 1BE8; X-ray; 2.20 A; A=106-379.
DR   PDB; 1BFK; X-ray; 2.30 A; A=106-379.
DR   PDB; 1BFU; X-ray; 2.20 A; A=106-379.
DR   PDB; 1C3L; X-ray; 2.16 A; A=106-379.
DR   PDB; 1CSE; X-ray; 1.20 A; E=106-379.
DR   PDB; 1OYV; X-ray; 2.50 A; A/B=106-379.
DR   PDB; 1R0R; X-ray; 1.10 A; E=106-379.
DR   PDB; 1SBC; X-ray; 2.50 A; A=106-379.
DR   PDB; 1SCA; X-ray; 2.00 A; A=106-379.
DR   PDB; 1SCB; X-ray; 2.30 A; A=106-379.
DR   PDB; 1SCD; X-ray; 2.30 A; A=106-379.
DR   PDB; 1SCN; X-ray; 1.90 A; E=106-379.
DR   PDB; 1SEL; X-ray; 2.00 A; A/B=106-379.
DR   PDB; 1VSB; X-ray; 2.10 A; A=106-379.
DR   PDB; 1YU6; X-ray; 1.55 A; A/B=105-379.
DR   PDB; 2SEC; X-ray; 1.80 A; E=106-379.
DR   PDB; 2WUV; X-ray; 2.24 A; A=106-379.
DR   PDB; 2WUW; X-ray; 2.23 A; E=106-379.
DR   PDB; 3UNX; X-ray; 1.26 A; A=106-379.
DR   PDB; 3VSB; X-ray; 2.60 A; A=106-379.
DR   PDB; 4C3U; X-ray; 2.29 A; A=106-379.
DR   PDB; 4C3V; X-ray; 2.26 A; A=106-379.
DR   PDB; 6DWQ; X-ray; 1.27 A; A=106-379.
DR   AlphaFoldDB; P00780; -.
DR   MDposit; P00780; -.
DR   PCDDB; P00780; -.
DR   SMR; P00780; -.
DR   IntAct; P00780; 2.
DR   MINT; P00780; -.
DR   BindingDB; P00780; -.
DR   ChEMBL; CHEMBL4299; -.
DR   DrugBank; DB03316; 1,4-Dioxane.
DR   DrugBank; DB03607; [(1R)-1-acetamido-2-(4-chlorophenyl)ethyl]-[(2S)-2-amino-3-hydroxy-3-oxo-propoxy]-dihydroxy-boron.
DR   DrugBank; DB02560; [(1S)-1-acetamido-2-(4-chlorophenyl)ethyl]-[(2S)-2-amino-3-hydroxy-3-oxo-propoxy]-dihydroxy-boron.
DR   DrugBank; DB02677; D-naphthyl-1-acetamido boronic acid alanine.
DR   DrugBank; DB01942; Formic acid.
DR   DrugBank; DB03290; L-naphthyl-1-acetamido boronic acid alanine.
DR   Allergome; 8254; Bac li Subtilisin.
DR   MEROPS; I09.001; -.
DR   MEROPS; S08.001; -.
DR   MetOSite; P00780; -.
DR   BRENDA; 3.4.21.62; 669.
DR   SABIO-RK; P00780; -.
DR   EvolutionaryTrace; P00780; -.
DR   GO; GO:0005576; C:extracellular region; EXP:UniProtKB.
DR   GO; GO:0004252; F:serine-type endopeptidase activity; EXP:UniProtKB.
DR   GO; GO:0006508; P:proteolysis; IEA:InterPro.
DR   CDD; cd07477; Peptidases_S8_Subtilisin_subset; 1.
DR   Gene3D; 3.30.70.80; Peptidase S8 propeptide/proteinase inhibitor I9; 1.
DR   Gene3D; 3.40.50.200; Peptidase S8/S53 domain; 1.
DR   InterPro; IPR000209; Peptidase_S8/S53_dom.
DR   InterPro; IPR036852; Peptidase_S8/S53_dom_sf.
DR   InterPro; IPR023827; Peptidase_S8_Asp-AS.
DR   InterPro; IPR022398; Peptidase_S8_His-AS.
DR   InterPro; IPR023828; Peptidase_S8_Ser-AS.
DR   InterPro; IPR050131; Peptidase_S8_subtilisin-like.
DR   InterPro; IPR015500; Peptidase_S8_subtilisin-rel.
DR   InterPro; IPR010259; S8pro/Inhibitor_I9.
DR   InterPro; IPR037045; S8pro/Inhibitor_I9_sf.
DR   InterPro; IPR034202; Subtilisin_Carlsberg-like.
DR   PANTHER; PTHR43806:SF11; CEREVISIN-RELATED; 1.
DR   PANTHER; PTHR43806; PEPTIDASE S8; 1.
DR   Pfam; PF05922; Inhibitor_I9; 1.
DR   Pfam; PF00082; Peptidase_S8; 1.
DR   PRINTS; PR00723; SUBTILISIN.
DR   SUPFAM; SSF54897; Protease propeptides/inhibitors; 1.
DR   SUPFAM; SSF52743; Subtilisin-like; 1.
DR   PROSITE; PS51892; SUBTILASE; 1.
DR   PROSITE; PS00136; SUBTILASE_ASP; 1.
DR   PROSITE; PS00137; SUBTILASE_HIS; 1.
DR   PROSITE; PS00138; SUBTILASE_SER; 1.
DR   PDBsum; 1AF4; -.
DR   PDBsum; 1AV7; -.
DR   PDBsum; 1AVT; -.
DR   PDBsum; 1BE6; -.
DR   PDBsum; 1BE8; -.
DR   PDBsum; 1BFK; -.
DR   PDBsum; 1BFU; -.
DR   PDBsum; 1C3L; -.
DR   PDBsum; 1CSE; -.
DR   PDBsum; 1OYV; -.
DR   PDBsum; 1R0R; -.
DR   PDBsum; 1SBC; -.
DR   PDBsum; 1SCA; -.
DR   PDBsum; 1SCB; -.
DR   PDBsum; 1SCD; -.
DR   PDBsum; 1SCN; -.
DR   PDBsum; 1SEL; -.
DR   PDBsum; 1VSB; -.
DR   PDBsum; 1YU6; -.
DR   PDBsum; 2SEC; -.
DR   PDBsum; 2WUV; -.
DR   PDBsum; 2WUW; -.
DR   PDBsum; 3UNX; -.
DR   PDBsum; 3VSB; -.
DR   PDBsum; 4C3U; -.
DR   PDBsum; 4C3V; -.
DR   PDBsum; 6DWQ; -.
PE   1: Evidence at protein level;
KW   3D-structure; Calcium; Direct protein sequencing; Hydrolase; Metal-binding;
KW   Protease; Secreted; Serine protease; Signal; Zymogen.
FT   SIGNAL          1..29
FT                   /evidence="ECO:0000255"
FT   PROPEP          30..105
FT                   /evidence="ECO:0000269|PubMed:4967581"
FT                   /id="PRO_0000027179"
FT   CHAIN           106..379
FT                   /note="Subtilisin Carlsberg"
FT                   /evidence="ECO:0000269|PubMed:4967581"
FT                   /id="PRO_0000027180"
FT   DOMAIN          44..102
FT                   /note="Inhibitor I9"
FT                   /evidence="ECO:0000255"
FT   DOMAIN          110..378
FT                   /note="Peptidase S8"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT   ACT_SITE        137
FT                   /note="Charge relay system"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT   ACT_SITE        168
FT                   /note="Charge relay system"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT   ACT_SITE        325
FT                   /note="Charge relay system"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT   BINDING         107
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000269|PubMed:8512925"
FT   BINDING         146
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000269|PubMed:8512925"
FT   BINDING         179
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000269|PubMed:8512925"
FT   BINDING         181
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000269|PubMed:8512925"
FT   BINDING         183
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000269|PubMed:8512925"
FT   BINDING         185
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="1"
FT                   /evidence="ECO:0000269|PubMed:8512925"
FT   BINDING         273
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="2"
FT                   /evidence="ECO:0000269|PubMed:8512925"
FT   BINDING         275
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="2"
FT                   /evidence="ECO:0000269|PubMed:8512925"
FT   BINDING         278
FT                   /ligand="Ca(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29108"
FT                   /ligand_label="2"
FT                   /evidence="ECO:0000269|PubMed:8512925"
FT   CONFLICT        207
FT                   /note="S -> T (in Ref. 1; CAB56500)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        233
FT                   /note="A -> P (in Ref. 1; CAB56500)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        249
FT                   /note="K -> R (in Ref. 1; CAB56500 and 3; AA sequence)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        262..265
FT                   /note="SSGN -> NSGS (in Ref. 3; AA sequence)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        316
FT                   /note="N -> S (in Ref. 1; CAB56500)"
FT                   /evidence="ECO:0000305"
FT   HELIX           112..115
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   HELIX           118..124
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          132..138
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          149..154
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          156..158
FT                   /evidence="ECO:0007829|PDB:1SBC"
FT   STRAND          161..163
FT                   /evidence="ECO:0007829|PDB:1SBC"
FT   STRAND          165..167
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   HELIX           168..177
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          180..184
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          192..198
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          204..206
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   HELIX           208..220
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          224..228
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          230..234
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   HELIX           237..248
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          252..256
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   TURN            272..274
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          278..284
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          300..305
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          307..313
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   TURN            314..316
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   STRAND          317..321
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   HELIX           324..341
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   HELIX           347..356
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   HELIX           364..367
FT                   /evidence="ECO:0007829|PDB:1R0R"
FT   HELIX           374..377
FT                   /evidence="ECO:0007829|PDB:1R0R"
SQ   SEQUENCE   379 AA;  38867 MW;  38432C04FFA048C1 CRC64;
     MMRKKSFWLG MLTAFMLVFT MAFSDSASAA QPAKNVEKDY IVGFKSGVKT ASVKKDIIKE
     SGGKVDKQFR IINAAKAKLD KEALKEVKND PDVAYVEEDH VAHALAQTVP YGIPLIKADK
     VQAQGFKGAN VKVAVLDTGI QASHPDLNVV GGASFVAGEA YNTDGNGHGT HVAGTVAALD
     NTTGVLGVAP SVSLYAVKVL NSSGSGSYSG IVSGIEWATT NGMDVINMSL GGASGSTAMK
     QAVDNAYAKG VVVVAAAGNS GSSGNTNTIG YPAKYDSVIA VGAVDSNSNR ASFSSVGAEL
     EVMAPGAGVY STYPTNTYAT LNGTSMASPH VAGAAALILS KHPNLSASQV RNRLSSTATY
     LGSSFYYGKG LINVEAAAQ
//