CAH_THEMA
ID CAH_THEMA Reviewed; 325 AA.
AC Q9WXT2; G4FGZ2;
DT 31-OCT-2012, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1999, sequence version 1.
DT 03-AUG-2022, entry version 119.
DE RecName: Full=Cephalosporin-C deacetylase {ECO:0000305};
DE EC=3.1.1.41 {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095};
DE AltName: Full=Acetylxylan esterase;
DE EC=3.1.1.72 {ECO:0000269|PubMed:21255309};
GN Name=axeA {ECO:0000303|PubMed:21255309}; OrderedLocusNames=TM_0077;
OS Thermotoga maritima (strain ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826
OS / MSB8).
OC Bacteria; Thermotogae; Thermotogales; Thermotogaceae; Thermotoga.
OX NCBI_TaxID=243274;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 / MSB8;
RX PubMed=10360571; DOI=10.1038/20601;
RA Nelson K.E., Clayton R.A., Gill S.R., Gwinn M.L., Dodson R.J., Haft D.H.,
RA Hickey E.K., Peterson J.D., Nelson W.C., Ketchum K.A., McDonald L.A.,
RA Utterback T.R., Malek J.A., Linher K.D., Garrett M.M., Stewart A.M.,
RA Cotton M.D., Pratt M.S., Phillips C.A., Richardson D.L., Heidelberg J.F.,
RA Sutton G.G., Fleischmann R.D., Eisen J.A., White O., Salzberg S.L.,
RA Smith H.O., Venter J.C., Fraser C.M.;
RT "Evidence for lateral gene transfer between Archaea and Bacteria from
RT genome sequence of Thermotoga maritima.";
RL Nature 399:323-329(1999).
RN [2] {ECO:0000305}
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RC STRAIN=ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 / MSB8
RC {ECO:0000269|PubMed:21255309};
RX PubMed=21255309; DOI=10.1111/j.1751-7915.2009.00150.x;
RA Drzewiecki K., Angelov A., Ballschmiter M., Tiefenbach K.J., Sterner R.,
RA Liebl W.;
RT "Hyperthermostable acetyl xylan esterase.";
RL Microb. Biotechnol. 3:84-92(2010).
RN [3] {ECO:0000305}
RP FUNCTION, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP MUTAGENESIS OF GLN-88; TYR-92; TRP-102; TRP-105; TRP-124; SER-188; PHE-213;
RP PRO-228 AND ILE-276.
RX PubMed=22659119; DOI=10.1016/j.bbapap.2012.05.009;
RA Hedge M.K., Gehring A.M., Adkins C.T., Weston L.A., Lavis L.D.,
RA Johnson R.J.;
RT "The structural basis for the narrow substrate specificity of an acetyl
RT esterase from Thermotoga maritima.";
RL Biochim. Biophys. Acta 1824:1024-1030(2012).
RN [4] {ECO:0007744|PDB:1VLQ, ECO:0007744|PDB:3M81, ECO:0007744|PDB:3M82, ECO:0007744|PDB:3M83}
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) IN COMPLEX WITH PMSF AND DIETHYL
RP PHOSPHONATE, FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND SUBUNIT.
RC STRAIN=ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 / MSB8
RC {ECO:0000269|PubMed:22411095};
RX PubMed=22411095; DOI=10.1002/prot.24041;
RA Levisson M., Han G.W., Deller M.C., Xu Q., Biely P., Hendriks S.,
RA Ten Eyck L.F., Flensburg C., Roversi P., Miller M.D., McMullan D.,
RA von Delft F., Kreusch A., Deacon A.M., van der Oost J., Lesley S.A.,
RA Elsliger M.A., Kengen S.W., Wilson I.A.;
RT "Functional and structural characterization of a thermostable acetyl
RT esterase from Thermotoga maritima.";
RL Proteins 80:1545-1559(2012).
CC -!- FUNCTION: Esterase that removes acetyl groups from a number of O-
CC acetylated small substrates, such as acetylated xylose, short xylo-
CC oligosaccharides and cephalosporin C. Has no activity towards polymeric
CC acetylated xylan, 4-methylumbelliferyl acetate or alpha-naphthyl
CC acetate. Able to catalyze rapid hydrolysis of a range of substrates
CC preferably with acetate groups, independent of the alcohol moiety.
CC Exhibits a narrow selectivity for short chain acyl esters (C2-C3).
CC Displays broad substrate specificity by hydrolyzing acetate at 2, 3,
CC and 4 positions of 4-nitrophenyl-beta-D-xylopyranoside (pNP-Xyl) with
CC similar efficiency. Cannot cleave amide linkages.
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Deacetylation of xylans and xylo-oligosaccharides.;
CC EC=3.1.1.72; Evidence={ECO:0000269|PubMed:21255309,
CC ECO:0000269|PubMed:22411095};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=cephalosporin C + H2O = acetate + deacetylcephalosporin C +
CC H(+); Xref=Rhea:RHEA:22596, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:30089, ChEBI:CHEBI:57511, ChEBI:CHEBI:58366; EC=3.1.1.41;
CC Evidence={ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095};
CC -!- ACTIVITY REGULATION: Activity stimulated by up to 40% in the presence
CC of divalent cations such as BaCl(2), CaCl(2), MgCl(2) and MnCl(2) at 3
CC mM concentration, but inhibited by 82-85% in the presence of CdCl(2)
CC and ZnCl(2) at 3 mM concentration. {ECO:0000269|PubMed:21255309}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.12 mM for p-nitrophenyl-acetate {ECO:0000269|PubMed:21255309,
CC ECO:0000269|PubMed:22411095, ECO:0000269|PubMed:22659119};
CC KM=6.05 mM for glucose penta-acetate {ECO:0000269|PubMed:21255309,
CC ECO:0000269|PubMed:22411095, ECO:0000269|PubMed:22659119};
CC KM=4.18 mM for cephalosporin-C {ECO:0000269|PubMed:21255309,
CC ECO:0000269|PubMed:22411095, ECO:0000269|PubMed:22659119};
CC KM=20.8 mM for 7-aminocephalosporonic acid (7-ACA)
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC KM=760 uM for p-nitrophenyl acetate {ECO:0000269|PubMed:21255309,
CC ECO:0000269|PubMed:22411095, ECO:0000269|PubMed:22659119};
CC KM=3.7 uM for fluoroscein di(acetoxymethyl) ether
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC KM=0.97 uM for fluoroscein dipropyloxymethyl ether
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC KM=0.41 uM for fluoroscein dibutyloxymethyl ether
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC KM=1.0 uM for fluoroscein divaleryloxymethyl ether
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC KM=2.6 uM for fluoroscein dicaproyloxymethyl ether
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC KM=0.51 uM for fluoroscein dimethacryloxymethyl ether
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC KM=0.96 uM for fluoroscein dicyclobutylcarboxymethyl ether
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC KM=0.54 uM for fluoroscein dimethylcyclopropanecarboxymethyl ether
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC KM=0.185 mM for p-nitrophenyl-acetate {ECO:0000269|PubMed:21255309,
CC ECO:0000269|PubMed:22411095, ECO:0000269|PubMed:22659119};
CC KM=0.137 mM for p-nitrophenyl-propionate
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC KM=3.6 mM for 2-O-acetyl-pNP-Xyl {ECO:0000269|PubMed:21255309,
CC ECO:0000269|PubMed:22411095, ECO:0000269|PubMed:22659119};
CC KM=4.2 mM for 3-O-acetyl-pNP-Xyl {ECO:0000269|PubMed:21255309,
CC ECO:0000269|PubMed:22411095, ECO:0000269|PubMed:22659119};
CC KM=4.0 mM for 4-O-acetyl-pNP-Xyl {ECO:0000269|PubMed:21255309,
CC ECO:0000269|PubMed:22411095, ECO:0000269|PubMed:22659119};
CC Vmax=113.5 umol/min/mg enzyme with pNP-acetate as substrate
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC Vmax=366.8 umol/min/mg enzyme with glucose penta-acetate as substrate
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC Vmax=19.2 umol/min/mg enzyme with cephalosporin-C as substrate
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC pH dependence:
CC Optimum pH is 6.5. Half-maximal activity between pH 5.0-7.5.
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC Temperature dependence:
CC Optimum temperature is 90 degrees Celsius. Activity drops sharply
CC above 90 degrees Celsius. Stable up to 100-104 degrees Celsius.
CC {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095,
CC ECO:0000269|PubMed:22659119};
CC -!- SUBUNIT: Homohexamer, formed by a trimer of dimers. Also exists as a
CC homodimer. {ECO:0000269|PubMed:21255309, ECO:0000269|PubMed:22411095}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000250|UniProtKB:P94388}.
CC -!- SIMILARITY: Belongs to the carbohydrate esterase 7 family.
CC {ECO:0000255}.
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DR EMBL; AE000512; AAD35171.1; -; Genomic_DNA.
DR PIR; E72421; E72421.
DR RefSeq; NP_227893.1; NC_000853.1.
DR RefSeq; WP_004082599.1; NZ_CP011107.1.
DR PDB; 1VLQ; X-ray; 2.10 A; A/B/C/D/E/F/G/H/I/J/K/L=1-325.
DR PDB; 3M81; X-ray; 2.50 A; A/B/C/D/E/F=1-325.
DR PDB; 3M82; X-ray; 2.40 A; A/B/C/D/E/F=1-325.
DR PDB; 3M83; X-ray; 2.12 A; A/B/C/D/E/F=1-325.
DR PDB; 5FDF; X-ray; 1.76 A; A/B/C/D/E/F=1-325.
DR PDB; 5GMA; X-ray; 2.10 A; A/B/C/D/E/F=1-325.
DR PDB; 5HFN; X-ray; 2.75 A; A/B/C/D/E/F=1-325.
DR PDB; 5JIB; X-ray; 1.86 A; A/B/C/D/E/F=1-325.
DR PDBsum; 1VLQ; -.
DR PDBsum; 3M81; -.
DR PDBsum; 3M82; -.
DR PDBsum; 3M83; -.
DR PDBsum; 5FDF; -.
DR PDBsum; 5GMA; -.
DR PDBsum; 5HFN; -.
DR PDBsum; 5JIB; -.
DR AlphaFoldDB; Q9WXT2; -.
DR SMR; Q9WXT2; -.
DR STRING; 243274.THEMA_04420; -.
DR ESTHER; thema-TM0077; Acetyl-esterase_deacetylase.
DR EnsemblBacteria; AAD35171; AAD35171; TM_0077.
DR KEGG; tma:TM0077; -.
DR eggNOG; COG3458; Bacteria.
DR InParanoid; Q9WXT2; -.
DR OMA; VHFAARG; -.
DR OrthoDB; 721431at2; -.
DR BRENDA; 3.1.1.41; 6331.
DR EvolutionaryTrace; Q9WXT2; -.
DR Proteomes; UP000008183; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0046555; F:acetylxylan esterase activity; IDA:UniProtKB.
DR GO; GO:0005509; F:calcium ion binding; IDA:UniProtKB.
DR GO; GO:0052689; F:carboxylic ester hydrolase activity; IBA:GO_Central.
DR GO; GO:0047739; F:cephalosporin-C deacetylase activity; IDA:UniProtKB.
DR GO; GO:0030245; P:cellulose catabolic process; IEA:UniProtKB-KW.
DR GO; GO:1901266; P:cephalosporin C metabolic process; IDA:UniProtKB.
DR GO; GO:0005976; P:polysaccharide metabolic process; IDA:UniProtKB.
DR GO; GO:0045491; P:xylan metabolic process; IDA:UniProtKB.
DR Gene3D; 3.40.50.1820; -; 1.
DR InterPro; IPR029058; AB_hydrolase.
DR InterPro; IPR008391; AXE1_dom.
DR InterPro; IPR039069; CE7.
DR PANTHER; PTHR40111; PTHR40111; 1.
DR Pfam; PF05448; AXE1; 1.
DR SUPFAM; SSF53474; SSF53474; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Calcium; Carbohydrate metabolism; Cellulose degradation;
KW Cytoplasm; Hydrolase; Metal-binding; Polysaccharide degradation;
KW Reference proteome; Serine esterase.
FT CHAIN 1..325
FT /note="Cephalosporin-C deacetylase"
FT /id="PRO_0000419766"
FT ACT_SITE 188
FT /note="Nucleophile"
FT /evidence="ECO:0000269|PubMed:22411095"
FT ACT_SITE 274
FT /note="Charge relay system"
FT /evidence="ECO:0000250|UniProtKB:P94388"
FT ACT_SITE 303
FT /note="Charge relay system"
FT /evidence="ECO:0000250|UniProtKB:P94388"
FT BINDING 92
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:P94388"
FT BINDING 188
FT /ligand="paraoxon"
FT /ligand_id="ChEBI:CHEBI:27827"
FT /ligand_note="inhibitor"
FT /note="covalent"
FT /evidence="ECO:0000269|PubMed:22411095"
FT MUTAGEN 88
FT /note="Q->A: Minimal change in catalytic efficiency toward
FT acetate substrates."
FT /evidence="ECO:0000269|PubMed:22659119"
FT MUTAGEN 92
FT /note="Y->A: Reduction in catalytic efficiency towards
FT fluoroscein diacetoxymethyl ether; when associated with A-
FT 228."
FT /evidence="ECO:0000269|PubMed:22659119"
FT MUTAGEN 102
FT /note="W->A: Minimal change in catalytic efficiency toward
FT the acetate substrates."
FT /evidence="ECO:0000269|PubMed:22659119"
FT MUTAGEN 105
FT /note="W->A: Minimal change in catalytic efficiency toward
FT the acetate substrates."
FT /evidence="ECO:0000269|PubMed:22659119"
FT MUTAGEN 124
FT /note="W->A: Minimal change in catalytic efficiency toward
FT the acetate substrates."
FT /evidence="ECO:0000269|PubMed:22659119"
FT MUTAGEN 188
FT /note="S->A: Activity abolished towards fluoroscein
FT diacetoxymethyl ether."
FT /evidence="ECO:0000269|PubMed:22659119"
FT MUTAGEN 213
FT /note="F->A: Reduction in catalytic efficiency towards
FT fluoroscein diacetoxymethyl ether; when associated with A-
FT 228."
FT /evidence="ECO:0000269|PubMed:22659119"
FT MUTAGEN 228
FT /note="P->A: Reduces catalytic activity by 33-fold against
FT fluoroscein diacetoxymethyl ether; 8-fold decrease in
FT activity against dimethacryloxymethyl ether; 4-fold
FT increase in catalytic efficiency toward
FT dicyclobutylcarboxymethyl ether. Reduction in catalytic
FT efficiency towards fluoroscein diacetoxymethyl ether; when
FT associated with A-92; A-213 or A-276."
FT /evidence="ECO:0000269|PubMed:22659119"
FT MUTAGEN 276
FT /note="I->A: Greater than 10-fold reduction in catalytic
FT efficiency toward the acetate substrates. Reduction in
FT catalytic efficiency towards fluoroscein diacetoxymethyl
FT ether; when associated with A-228."
FT /evidence="ECO:0000269|PubMed:22659119"
FT HELIX 8..11
FT /evidence="ECO:0007829|PDB:5FDF"
FT HELIX 24..36
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 43..46
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 52..62
FT /evidence="ECO:0007829|PDB:5FDF"
FT HELIX 64..66
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 68..76
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 81..89
FT /evidence="ECO:0007829|PDB:5FDF"
FT HELIX 99..102
FT /evidence="ECO:0007829|PDB:5FDF"
FT HELIX 104..107
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 111..115
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 123..125
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 134..137
FT /evidence="ECO:0007829|PDB:1VLQ"
FT STRAND 142..144
FT /evidence="ECO:0007829|PDB:5FDF"
FT TURN 145..150
FT /evidence="ECO:0007829|PDB:5FDF"
FT TURN 152..154
FT /evidence="ECO:0007829|PDB:5FDF"
FT HELIX 156..172
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 177..187
FT /evidence="ECO:0007829|PDB:5FDF"
FT HELIX 189..200
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 205..211
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 213..215
FT /evidence="ECO:0007829|PDB:5HFN"
FT HELIX 217..223
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 225..227
FT /evidence="ECO:0007829|PDB:5GMA"
FT HELIX 229..238
FT /evidence="ECO:0007829|PDB:5FDF"
FT HELIX 240..242
FT /evidence="ECO:0007829|PDB:5HFN"
FT HELIX 243..251
FT /evidence="ECO:0007829|PDB:5FDF"
FT HELIX 255..259
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 266..271
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 275..277
FT /evidence="ECO:0007829|PDB:5FDF"
FT HELIX 279..288
FT /evidence="ECO:0007829|PDB:5FDF"
FT STRAND 291..298
FT /evidence="ECO:0007829|PDB:5FDF"
FT TURN 303..306
FT /evidence="ECO:0007829|PDB:5FDF"
FT HELIX 307..322
FT /evidence="ECO:0007829|PDB:5FDF"
SQ SEQUENCE 325 AA; 37156 MW; AF2879D86680A3C4 CRC64;
MAFFDLPLEE LKKYRPERYE EKDFDEFWEE TLAESEKFPL DPVFERMESH LKTVEAYDVT
FSGYRGQRIK GWLLVPKLEE EKLPCVVQYI GYNGGRGFPH DWLFWPSMGY ICFVMDTRGQ
GSGWLKGDTP DYPEGPVDPQ YPGFMTRGIL DPRTYYYRRV FTDAVRAVEA AASFPQVDQE
RIVIAGGSQG GGIALAVSAL SKKAKALLCD VPFLCHFRRA VQLVDTHPYA EITNFLKTHR
DKEEIVFRTL SYFDGVNFAA RAKIPALFSV GLMDNICPPS TVFAAYNYYA GPKEIRIYPY
NNHEGGGSFQ AVEQVKFLKK LFEKG
