ID   ESTA_BACSU              Reviewed;         212 AA.
AC   P37957; O34644; Q2XU59;
DT   01-OCT-1994, integrated into UniProtKB/Swiss-Prot.
DT   30-MAY-2000, sequence version 2.
DT   27-MAR-2024, entry version 154.
DE   RecName: Full=Lipase EstA;
DE   AltName: Full=Lipase A;
DE            EC=3.1.1.3 {ECO:0000269|PubMed:11029590, ECO:0000269|PubMed:8396026};
DE   AltName: Full=Triacylglycerol lipase;
DE   Flags: Precursor;
GN   Name=estA;
GN   Synonyms=lip {ECO:0000303|PubMed:1320940}, lipA
GN   {ECO:0000303|PubMed:11029590}; OrderedLocusNames=BSU02700;
OS   Bacillus subtilis (strain 168).
OC   Bacteria; Bacillota; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX   NCBI_TaxID=224308;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND FUNCTION.
RC   STRAIN=168;
RX   PubMed=1320940; DOI=10.1016/0167-4781(92)90023-s;
RA   Dartois V., Baulard A., Schanck K., Colson C.;
RT   "Cloning, nucleotide sequence and expression in Escherichia coli of a
RT   lipase gene from Bacillus subtilis 168.";
RL   Biochim. Biophys. Acta 1131:253-260(1992).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC   STRAIN=168;
RX   PubMed=9274031; DOI=10.1099/00221287-143-8-2775;
RA   Kumano M., Tamakoshi A., Yamane K.;
RT   "A 32 kb nucleotide sequence from the region of the lincomycin-resistance
RT   gene (22 degrees-25 degrees) of the Bacillus subtilis chromosome and
RT   identification of the site of the lin-2 mutation.";
RL   Microbiology 143:2775-2782(1997).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RA   Peerzada K., Johri S., Rasool S., Qazi G.N.;
RT   "Molecular characterization of a lipase from a strain of Bacillus
RT   subtilis.";
RL   Submitted (OCT-2005) to the EMBL/GenBank/DDBJ databases.
RN   [4]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=168;
RX   PubMed=9384377; DOI=10.1038/36786;
RA   Kunst F., Ogasawara N., Moszer I., Albertini A.M., Alloni G., Azevedo V.,
RA   Bertero M.G., Bessieres P., Bolotin A., Borchert S., Borriss R.,
RA   Boursier L., Brans A., Braun M., Brignell S.C., Bron S., Brouillet S.,
RA   Bruschi C.V., Caldwell B., Capuano V., Carter N.M., Choi S.-K.,
RA   Codani J.-J., Connerton I.F., Cummings N.J., Daniel R.A., Denizot F.,
RA   Devine K.M., Duesterhoeft A., Ehrlich S.D., Emmerson P.T., Entian K.-D.,
RA   Errington J., Fabret C., Ferrari E., Foulger D., Fritz C., Fujita M.,
RA   Fujita Y., Fuma S., Galizzi A., Galleron N., Ghim S.-Y., Glaser P.,
RA   Goffeau A., Golightly E.J., Grandi G., Guiseppi G., Guy B.J., Haga K.,
RA   Haiech J., Harwood C.R., Henaut A., Hilbert H., Holsappel S., Hosono S.,
RA   Hullo M.-F., Itaya M., Jones L.-M., Joris B., Karamata D., Kasahara Y.,
RA   Klaerr-Blanchard M., Klein C., Kobayashi Y., Koetter P., Koningstein G.,
RA   Krogh S., Kumano M., Kurita K., Lapidus A., Lardinois S., Lauber J.,
RA   Lazarevic V., Lee S.-M., Levine A., Liu H., Masuda S., Mauel C.,
RA   Medigue C., Medina N., Mellado R.P., Mizuno M., Moestl D., Nakai S.,
RA   Noback M., Noone D., O'Reilly M., Ogawa K., Ogiwara A., Oudega B.,
RA   Park S.-H., Parro V., Pohl T.M., Portetelle D., Porwollik S.,
RA   Prescott A.M., Presecan E., Pujic P., Purnelle B., Rapoport G., Rey M.,
RA   Reynolds S., Rieger M., Rivolta C., Rocha E., Roche B., Rose M., Sadaie Y.,
RA   Sato T., Scanlan E., Schleich S., Schroeter R., Scoffone F., Sekiguchi J.,
RA   Sekowska A., Seror S.J., Serror P., Shin B.-S., Soldo B., Sorokin A.,
RA   Tacconi E., Takagi T., Takahashi H., Takemaru K., Takeuchi M.,
RA   Tamakoshi A., Tanaka T., Terpstra P., Tognoni A., Tosato V., Uchiyama S.,
RA   Vandenbol M., Vannier F., Vassarotti A., Viari A., Wambutt R., Wedler E.,
RA   Wedler H., Weitzenegger T., Winters P., Wipat A., Yamamoto H., Yamane K.,
RA   Yasumoto K., Yata K., Yoshida K., Yoshikawa H.-F., Zumstein E.,
RA   Yoshikawa H., Danchin A.;
RT   "The complete genome sequence of the Gram-positive bacterium Bacillus
RT   subtilis.";
RL   Nature 390:249-256(1997).
RN   [5]
RP   FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP   PROPERTIES, SUBCELLULAR LOCATION, INDUCTION, AND N-TERMINAL END.
RC   STRAIN=168;
RX   PubMed=8396026; DOI=10.1111/j.1432-1033.1993.tb18127.x;
RA   Lesuisse E., Schanck K., Colson C.;
RT   "Purification and preliminary characterization of the extracellular lipase
RT   of Bacillus subtilis 168, an extremely basic pH-tolerant enzyme.";
RL   Eur. J. Biochem. 216:155-160(1993).
RN   [6]
RP   FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, AND BIOPHYSICOCHEMICAL
RP   PROPERTIES.
RC   STRAIN=168 / BCL 1050;
RX   PubMed=11029590; DOI=10.1046/j.1432-1327.2000.01736.x;
RA   Eggert T., Pencreac'h G., Douchet I., Verger R., Jaeger K.-E.;
RT   "A novel extracellular esterase from Bacillus subtilis and its conversion
RT   to a monoacylglycerol hydrolase.";
RL   Eur. J. Biochem. 267:6459-6469(2000).
RN   [7]
RP   PROBABLE ACTIVE SITE, SUBCELLULAR LOCATION, AND INDUCTION.
RC   STRAIN=168 / BCL 1050;
RX   PubMed=11583117; DOI=10.1016/s0014-5793(01)02665-5;
RA   Eggert T., van Pouderoyen G., Dijkstra B.W., Jaeger K.-E.;
RT   "Lipolytic enzymes LipA and LipB from Bacillus subtilis differ in
RT   regulation of gene expression, biochemical properties, and three-
RT   dimensional structure.";
RL   FEBS Lett. 502:89-92(2001).
RN   [8] {ECO:0007744|PDB:1I6W}
RP   X-RAY CRYSTALLOGRAPHY (1.5 ANGSTROMS) OF 32-212, PROBABLE ACTIVE SITE, AND
RP   PROBABLE SUBUNIT.
RC   STRAIN=168;
RX   PubMed=11491291; DOI=10.1006/jmbi.2001.4659;
RA   van Pouderoyen G., Eggert T., Jaeger K.-E., Dijkstra B.W.;
RT   "The crystal structure of Bacillus subtilis lipase: a minimal alpha/beta
RT   hydrolase fold enzyme.";
RL   J. Mol. Biol. 309:215-226(2001).
RN   [9] {ECO:0007744|PDB:1ISP}
RP   X-RAY CRYSTALLOGRAPHY (1.3 ANGSTROMS) OF 32-212, PROBABLE ACTIVE SITE, AND
RP   SUBUNIT.
RC   STRAIN=168;
RX   PubMed=12077437; DOI=10.1107/s090744490200714x;
RA   Kawasaki K., Kondo H., Suzuki M., Ohgiya S., Tsuda S.;
RT   "Alternate conformations observed in catalytic serine of Bacillus subtilis
RT   lipase determined at 1.3 A resolution.";
RL   Acta Crystallogr. D 58:1168-1174(2002).
CC   -!- FUNCTION: Active toward triacylglycerides with a preference for esters
CC       with C8:0 acyl groups; barely active on C18:1 or C18:4 substrates.
CC       Active against p-nitrophenylesters with fatty acid chain lengths from
CC       C6 to C18. {ECO:0000269|PubMed:11029590, ECO:0000269|PubMed:8396026,
CC       ECO:0000305|PubMed:1320940}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=a triacylglycerol + H2O = a diacylglycerol + a fatty acid +
CC         H(+); Xref=Rhea:RHEA:12044, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:17855, ChEBI:CHEBI:18035, ChEBI:CHEBI:28868; EC=3.1.1.3;
CC         Evidence={ECO:0000269|PubMed:11029590, ECO:0000269|PubMed:8396026};
CC   -!- ACTIVITY REGULATION: Strongly inhibited when incubated with the serine
CC       reagent phenylmethylsulfonyl fluoride. Activated by the addition of
CC       calcium to the reaction mixture. When calcium was incubated with the
CC       lipase but not added to the reaction mixture, its effect is lower but
CC       still observable. Magnesium, manganese and strontium are not able to
CC       replace calcium with full retention of activity.
CC       {ECO:0000269|PubMed:8396026}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       pH dependence:
CC         Optimum pH is 10 (PubMed:11029590). The activity decreases strongly
CC         above pH 10.5 or below pH 6.5. The enzyme is remarkably stable at
CC         alkaline pH, showing maximum stability at pH 12 and retaining more
CC         than 65% of its activity after 24 hours at pH 13 (PubMed:8396026).
CC         {ECO:0000269|PubMed:11029590, ECO:0000269|PubMed:8396026};
CC       Temperature dependence:
CC         Optimum temperature is 35 degrees Celsius. Stable for at least 30
CC         minutes at 40 degrees Celsius. Virtually no activity remains after 30
CC         minutes at 55 degrees Celsius. {ECO:0000269|PubMed:11029590,
CC         ECO:0000269|PubMed:8396026};
CC   -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:11491291,
CC       ECO:0000269|PubMed:12077437}.
CC   -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:11583117,
CC       ECO:0000269|PubMed:8396026}.
CC   -!- INDUCTION: Maximally expressed in late exponential growth phase.
CC       Expression decreases rapidly in the stationary phase. Expressed in both
CC       rich and minimal media with glucose as carbon source (at protein
CC       level). {ECO:0000269|PubMed:11583117, ECO:0000269|PubMed:8396026}.
CC   -!- SIMILARITY: Belongs to the AB hydrolase superfamily. {ECO:0000305}.
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DR   EMBL; M74010; AAA22574.1; -; Genomic_DNA.
DR   EMBL; AB000617; BAA22231.1; -; Genomic_DNA.
DR   EMBL; DQ250714; ABB54395.1; -; Genomic_DNA.
DR   EMBL; AL009126; CAB12064.1; -; Genomic_DNA.
DR   PIR; S23934; S23934.
DR   RefSeq; NP_388152.1; NC_000964.3.
DR   RefSeq; WP_003246250.1; NZ_JNCM01000030.1.
DR   PDB; 1I6W; X-ray; 1.50 A; A/B=32-212.
DR   PDB; 1ISP; X-ray; 1.30 A; A=32-212.
DR   PDB; 1R4Z; X-ray; 1.80 A; A/B=32-212.
DR   PDB; 1R50; X-ray; 1.45 A; A/B=32-212.
DR   PDB; 1T2N; X-ray; 1.80 A; A=32-212.
DR   PDB; 1T4M; X-ray; 2.00 A; A=32-212.
DR   PDB; 2QXT; X-ray; 2.00 A; A/B=34-212.
DR   PDB; 2QXU; X-ray; 1.90 A; A/B/C/D/E/F/G/H=34-212.
DR   PDB; 3D2A; X-ray; 1.73 A; A=32-212.
DR   PDB; 3D2B; X-ray; 1.95 A; A/B=32-212.
DR   PDB; 3D2C; X-ray; 2.18 A; A/B/C/D/E/F/G/H/I/J/K/L=32-212.
DR   PDB; 3QMM; X-ray; 1.89 A; A/B=32-212.
DR   PDB; 3QZU; X-ray; 1.85 A; A/B=32-212.
DR   PDB; 5CRI; X-ray; 1.63 A; A/B=32-212.
DR   PDB; 5CT4; X-ray; 1.49 A; A/B=33-212.
DR   PDB; 5CT5; X-ray; 1.75 A; A/B=33-212.
DR   PDB; 5CT6; X-ray; 1.90 A; A/B=33-212.
DR   PDB; 5CT8; X-ray; 1.29 A; A=33-212.
DR   PDB; 5CT9; X-ray; 1.40 A; A=33-212.
DR   PDB; 5CTA; X-ray; 1.24 A; A=33-212.
DR   PDB; 5CUR; X-ray; 1.30 A; A=33-212.
DR   PDBsum; 1I6W; -.
DR   PDBsum; 1ISP; -.
DR   PDBsum; 1R4Z; -.
DR   PDBsum; 1R50; -.
DR   PDBsum; 1T2N; -.
DR   PDBsum; 1T4M; -.
DR   PDBsum; 2QXT; -.
DR   PDBsum; 2QXU; -.
DR   PDBsum; 3D2A; -.
DR   PDBsum; 3D2B; -.
DR   PDBsum; 3D2C; -.
DR   PDBsum; 3QMM; -.
DR   PDBsum; 3QZU; -.
DR   PDBsum; 5CRI; -.
DR   PDBsum; 5CT4; -.
DR   PDBsum; 5CT5; -.
DR   PDBsum; 5CT6; -.
DR   PDBsum; 5CT8; -.
DR   PDBsum; 5CT9; -.
DR   PDBsum; 5CTA; -.
DR   PDBsum; 5CUR; -.
DR   AlphaFoldDB; P37957; -.
DR   BMRB; P37957; -.
DR   SMR; P37957; -.
DR   STRING; 224308.BSU02700; -.
DR   DrugBank; DB08475; [(4R)-2,2-dimethyl-1,3-dioxolan-4-yl]methyl hydrogen hex-5-enylphosphonate.
DR   DrugBank; DB08548; [(4S)-2,2-dimethyl-1,3-dioxolan-4-yl]methyl hydrogen hex-5-enylphosphonate.
DR   ESTHER; bacsu-lip; Lipase_2.
DR   PaxDb; 224308-BSU02700; -.
DR   EnsemblBacteria; CAB12064; CAB12064; BSU_02700.
DR   GeneID; 938389; -.
DR   KEGG; bsu:BSU02700; -.
DR   PATRIC; fig|224308.179.peg.280; -.
DR   eggNOG; COG1075; Bacteria.
DR   InParanoid; P37957; -.
DR   OrthoDB; 503948at2; -.
DR   PhylomeDB; P37957; -.
DR   BioCyc; BSUB:BSU02700-MONOMER; -.
DR   SABIO-RK; P37957; -.
DR   EvolutionaryTrace; P37957; -.
DR   Proteomes; UP000001570; Chromosome.
DR   GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR   GO; GO:0016298; F:lipase activity; IBA:GO_Central.
DR   GO; GO:0004806; F:triglyceride lipase activity; IDA:CACAO.
DR   GO; GO:0016042; P:lipid catabolic process; IBA:GO_Central.
DR   Gene3D; 3.40.50.1820; alpha/beta hydrolase; 1.
DR   InterPro; IPR029058; AB_hydrolase.
DR   InterPro; IPR002918; Lipase_EstA/Esterase_EstB.
DR   PANTHER; PTHR32015; FASTING INDUCED LIPASE; 1.
DR   PANTHER; PTHR32015:SF1; FASTING INDUCED LIPASE; 1.
DR   Pfam; PF01674; Lipase_2; 1.
DR   SUPFAM; SSF53474; alpha/beta-Hydrolases; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Hydrolase; Lipid degradation; Lipid metabolism;
KW   Reference proteome; Secreted; Signal.
FT   SIGNAL          1..31
FT                   /evidence="ECO:0000305|PubMed:8396026"
FT   CHAIN           32..212
FT                   /note="Lipase EstA"
FT                   /id="PRO_0000017819"
FT   ACT_SITE        108
FT                   /note="Nucleophile"
FT                   /evidence="ECO:0000305|PubMed:11491291,
FT                   ECO:0000305|PubMed:11583117, ECO:0000305|PubMed:12077437"
FT   ACT_SITE        164
FT                   /note="Charge relay system"
FT                   /evidence="ECO:0000305|PubMed:11491291,
FT                   ECO:0000305|PubMed:11583117, ECO:0000305|PubMed:12077437"
FT   ACT_SITE        187
FT                   /note="Charge relay system"
FT                   /evidence="ECO:0000305|PubMed:11491291,
FT                   ECO:0000305|PubMed:11583117, ECO:0000305|PubMed:12077437"
FT   CONFLICT        29
FT                   /note="A -> V (in Ref. 3; ABB54395)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        51
FT                   /note="A -> G (in Ref. 3; ABB54395)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        60
FT                   /note="Q -> H (in Ref. 3; ABB54395)"
FT                   /evidence="ECO:0000305"
FT   CONFLICT        133
FT                   /note="L -> V (in Ref. 1; AAA22574 and 3; ABB54395)"
FT                   /evidence="ECO:0000305"
FT   STRAND          37..40
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   HELIX           47..50
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   HELIX           51..59
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   HELIX           64..66
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   STRAND          67..69
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   HELIX           79..97
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   STRAND          102..107
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   HELIX           110..120
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   HELIX           123..125
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   STRAND          127..134
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   HELIX           137..139
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   STRAND          155..161
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   STRAND          165..167
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   HELIX           169..172
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   STRAND          177..184
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   HELIX           189..191
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   HELIX           194..204
FT                   /evidence="ECO:0007829|PDB:5CTA"
FT   TURN            205..207
FT                   /evidence="ECO:0007829|PDB:5CTA"
SQ   SEQUENCE   212 AA;  22791 MW;  B8A70E027461188F CRC64;
     MKFVKRRIIA LVTILMLSVT SLFALQPSAK AAEHNPVVMV HGIGGASFNF AGIKSYLVSQ
     GWSRDKLYAV DFWDKTGTNY NNGPVLSRFV QKVLDETGAK KVDIVAHSMG GANTLYYIKN
     LDGGNKVANV VTLGGANRLT TGKALPGTDP NQKILYTSIY SSADMIVMNY LSRLDGARNV
     QIHGVGHIGL LYSSQVNSLI KEGLNGGGQN TN
//