GAD_SACS2
ID GAD_SACS2 Reviewed; 395 AA.
AC Q97U27;
DT 26-JUN-2013, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-2001, sequence version 1.
DT 03-AUG-2022, entry version 98.
DE RecName: Full=D-gluconate/D-galactonate dehydratase {ECO:0000303|PubMed:15474024};
DE Short=GAD {ECO:0000303|PubMed:15474024};
DE Short=GNAD {ECO:0000303|PubMed:15474024};
DE EC=4.2.1.140 {ECO:0000269|PubMed:15474024, ECO:0000269|PubMed:15509194};
DE EC=4.2.1.39 {ECO:0000269|PubMed:15474024, ECO:0000269|PubMed:15509194};
DE EC=4.2.1.6 {ECO:0000269|PubMed:15474024, ECO:0000269|PubMed:15509194};
GN Name=gad {ECO:0000303|PubMed:15474024};
GN Synonyms=gnaD {ECO:0000303|PubMed:15509194}; OrderedLocusNames=SSO3198;
OS Saccharolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)
OS (Sulfolobus solfataricus).
OC Archaea; Crenarchaeota; Thermoprotei; Sulfolobales; Sulfolobaceae;
OC Saccharolobus.
OX NCBI_TaxID=273057;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 35092 / DSM 1617 / JCM 11322 / P2;
RX PubMed=11427726; DOI=10.1073/pnas.141222098;
RA She Q., Singh R.K., Confalonieri F., Zivanovic Y., Allard G., Awayez M.J.,
RA Chan-Weiher C.C.-Y., Clausen I.G., Curtis B.A., De Moors A., Erauso G.,
RA Fletcher C., Gordon P.M.K., Heikamp-de Jong I., Jeffries A.C., Kozera C.J.,
RA Medina N., Peng X., Thi-Ngoc H.P., Redder P., Schenk M.E., Theriault C.,
RA Tolstrup N., Charlebois R.L., Doolittle W.F., Duguet M., Gaasterland T.,
RA Garrett R.A., Ragan M.A., Sensen C.W., Van der Oost J.;
RT "The complete genome of the crenarchaeon Sulfolobus solfataricus P2.";
RL Proc. Natl. Acad. Sci. U.S.A. 98:7835-7840(2001).
RN [2]
RP PROTEIN SEQUENCE OF 1-5, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, COFACTOR, SUBSTRATE SPECIFICITY, AND SUBUNIT.
RC STRAIN=ATCC 35092 / DSM 1617 / JCM 11322 / P2;
RX PubMed=15474024; DOI=10.1016/j.febslet.2004.08.074;
RA Lamble H.J., Milburn C.C., Taylor G.L., Hough D.W., Danson M.J.;
RT "Gluconate dehydratase from the promiscuous Entner-Doudoroff pathway in
RT Sulfolobus solfataricus.";
RL FEBS Lett. 576:133-136(2004).
RN [3]
RP PROTEIN SEQUENCE OF 1-10, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, COFACTOR, SUBSTRATE SPECIFICITY, SUBUNIT, AND NOMENCLATURE.
RC STRAIN=ATCC 35092 / DSM 1617 / JCM 11322 / P2;
RX PubMed=15509194; DOI=10.1042/bj20041053;
RA Kim S., Lee S.B.;
RT "Identification and characterization of Sulfolobus solfataricus D-gluconate
RT dehydratase: a key enzyme in the non-phosphorylated Entner-Doudoroff
RT pathway.";
RL Biochem. J. 387:271-280(2005).
CC -!- FUNCTION: Involved in the degradation of glucose and galactose via the
CC branched variant of the Entner-Doudoroff pathway. Catalyzes the
CC dehydration of gluconate to produce 2-keto-3-deoxygluconate (KDG). It
CC is also able to catalyze the dehydration of galactonate to produce 2-
CC keto-3-deoxygalactonate (KDGal). {ECO:0000269|PubMed:15474024,
CC ECO:0000269|PubMed:15509194}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=D-gluconate = 2-dehydro-3-deoxy-D-gluconate + H2O;
CC Xref=Rhea:RHEA:21612, ChEBI:CHEBI:15377, ChEBI:CHEBI:18391,
CC ChEBI:CHEBI:57990; EC=4.2.1.140;
CC Evidence={ECO:0000269|PubMed:15474024, ECO:0000269|PubMed:15509194};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=D-gluconate = 2-dehydro-3-deoxy-D-gluconate + H2O;
CC Xref=Rhea:RHEA:21612, ChEBI:CHEBI:15377, ChEBI:CHEBI:18391,
CC ChEBI:CHEBI:57990; EC=4.2.1.39;
CC Evidence={ECO:0000269|PubMed:15474024, ECO:0000269|PubMed:15509194};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=D-galactonate = 2-dehydro-3-deoxy-D-galactonate + H2O;
CC Xref=Rhea:RHEA:18649, ChEBI:CHEBI:12931, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:57989; EC=4.2.1.140;
CC Evidence={ECO:0000269|PubMed:15474024, ECO:0000269|PubMed:15509194};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=D-galactonate = 2-dehydro-3-deoxy-D-galactonate + H2O;
CC Xref=Rhea:RHEA:18649, ChEBI:CHEBI:12931, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:57989; EC=4.2.1.6; Evidence={ECO:0000269|PubMed:15474024,
CC ECO:0000269|PubMed:15509194};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:15474024, ECO:0000269|PubMed:15509194};
CC Name=Co(2+); Xref=ChEBI:CHEBI:48828;
CC Evidence={ECO:0000269|PubMed:15474024, ECO:0000269|PubMed:15509194};
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:15474024, ECO:0000269|PubMed:15509194};
CC Name=Ni(2+); Xref=ChEBI:CHEBI:49786;
CC Evidence={ECO:0000269|PubMed:15474024, ECO:0000269|PubMed:15509194};
CC Note=Binds 1 Mg(2+) ion per subunit. Can also use divalent metal ions
CC such as Co(2+), Mn(2+) and Ni(2+). {ECO:0000269|PubMed:15474024,
CC ECO:0000269|PubMed:15509194};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.45 mM for gluconate (at 78 degrees Celsius and pH 7)
CC {ECO:0000269|PubMed:15509194};
CC KM=0.81 mM for galactonate (at 70 degrees Celsius and pH 6)
CC {ECO:0000269|PubMed:15474024};
CC KM=1.57 mM for gluconate (at 70 degrees Celsius and pH 6)
CC {ECO:0000269|PubMed:15474024};
CC Vmax=0.15 umol/min/mg enzyme with gluconate as substrate (at 78
CC degrees Celsius and pH 7) {ECO:0000269|PubMed:15509194};
CC Note=kcat is 10.4 sec(-1) for gluconate and 1.08 sec(-1) for
CC galactonate (at 70 degrees Celsius and pH 6). kcat is 12.6 sec(-1)
CC for gluconate (at 78 degrees Celsius and pH 7).
CC {ECO:0000269|PubMed:15474024, ECO:0000269|PubMed:15509194};
CC pH dependence:
CC Optimum pH is between 6.5 and 7.5. {ECO:0000269|PubMed:15474024,
CC ECO:0000269|PubMed:15509194};
CC Temperature dependence:
CC Optimum temperature is between 80 and 100 degrees Celsius. At 80
CC degrees Celsius, the dehydratase is stable for over 2 hours, but at
CC 90 degrees Celsius its activity is decreased to less than 50% in 2
CC hours and its half-life diminishes to 120 min. At 100 degrees
CC Celsius, the enzyme has a half-life of less than 40 min. The activity
CC is almost undetectable below 60 degrees Celsius.
CC {ECO:0000269|PubMed:15474024, ECO:0000269|PubMed:15509194};
CC -!- PATHWAY: Carbohydrate acid metabolism; D-gluconate degradation.
CC {ECO:0000305}.
CC -!- SUBUNIT: Monomer or homooctamer. {ECO:0000269|PubMed:15474024,
CC ECO:0000269|PubMed:15509194}.
CC -!- MISCELLANEOUS: D-gluconate dehydratase activity is lost after
CC incubation with phosphatase, whereas no such decrease in activity is
CC observed when the enzymes are incubated in the same reaction mixtures
CC without phosphatase. Analysis of the amino acid sequence of D-gluconate
CC dehydratase indicate the presence of several putative phosphorylation
CC sites on serine, threonine and tyrosine residues.
CC {ECO:0000303|PubMed:15509194}.
CC -!- SIMILARITY: Belongs to the mandelate racemase/muconate lactonizing
CC enzyme family. GaD subfamily. {ECO:0000305}.
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DR EMBL; AE006641; AAK43295.1; -; Genomic_DNA.
DR PIR; H90504; H90504.
DR RefSeq; WP_009991686.1; NC_002754.1.
DR AlphaFoldDB; Q97U27; -.
DR SMR; Q97U27; -.
DR STRING; 273057.SSO3198; -.
DR EnsemblBacteria; AAK43295; AAK43295; SSO3198.
DR GeneID; 44128914; -.
DR KEGG; sso:SSO3198; -.
DR PATRIC; fig|273057.12.peg.3301; -.
DR eggNOG; arCOG01168; Archaea.
DR HOGENOM; CLU_030273_3_2_2; -.
DR InParanoid; Q97U27; -.
DR OMA; PRWCFLK; -.
DR PhylomeDB; Q97U27; -.
DR BRENDA; 4.2.1.140; 6163.
DR SABIO-RK; Q97U27; -.
DR UniPathway; UPA00792; -.
DR Proteomes; UP000001974; Chromosome.
DR GO; GO:0008869; F:galactonate dehydratase activity; IDA:UniProtKB.
DR GO; GO:0047929; F:gluconate dehydratase activity; IDA:UniProtKB.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0046177; P:D-gluconate catabolic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 3.20.20.120; -; 1.
DR Gene3D; 3.30.390.10; -; 1.
DR InterPro; IPR036849; Enolase-like_C_sf.
DR InterPro; IPR029017; Enolase-like_N.
DR InterPro; IPR029065; Enolase_C-like.
DR InterPro; IPR034599; Gluconate_dehydratase.
DR InterPro; IPR013342; Mandelate_racemase_C.
DR InterPro; IPR034593; Mandelate_racemase_DgoD-like.
DR InterPro; IPR013341; Mandelate_racemase_N_dom.
DR PANTHER; PTHR48080; PTHR48080; 1.
DR Pfam; PF13378; MR_MLE_C; 1.
DR Pfam; PF02746; MR_MLE_N; 1.
DR SFLD; SFLDF00008; gluconate_dehydratase; 1.
DR SMART; SM00922; MR_MLE; 1.
DR SUPFAM; SSF51604; SSF51604; 1.
DR SUPFAM; SSF54826; SSF54826; 1.
PE 1: Evidence at protein level;
KW Carbohydrate metabolism; Direct protein sequencing; Lyase; Magnesium;
KW Metal-binding; Reference proteome.
FT CHAIN 1..395
FT /note="D-gluconate/D-galactonate dehydratase"
FT /id="PRO_0000422650"
FT ACT_SITE 199
FT /note="Proton donor"
FT /evidence="ECO:0000250"
FT ACT_SITE 299
FT /note="Proton acceptor"
FT /evidence="ECO:0000250"
FT BINDING 197
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250"
FT BINDING 223
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250"
FT BINDING 249
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250"
FT SITE 272
FT /note="Increases basicity of active site His"
FT /evidence="ECO:0000250"
FT SITE 324
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000250"
SQ SEQUENCE 395 AA; 44729 MW; BA53EFD93F90A44D CRC64;
MRIREIEPIV LTSKEKGSAT WASIMIVTRV ITENGEVGYG EAVPTLRVIS VYNAIKQVSK
AYIGKEVEEV EKNYHEWYKQ DFYLARSFES ATAVSAIDIA SWDIIGKELG APIHKLLGGK
TRDRVPVYAN GWYQDCVTPE EFAEKAKDVV KMGYKALKFD PFGPYYDWID ERGLREAEER
VKAVREAVGD NVDILIEHHG RFNANSAIMI AKRLEKYNPG FMEEPVHHED VIGLRKYKAS
THLRVALGER LISEKETAFY VEEGLVNILQ PDLTNIGGVT VGRSVIKIAE ANDVEVAFHN
AFGSIQNAVE IQLSAVTQNL YLLENFYDWF PQWKRDLVYN ETPVEGGHVK VPYKPGLGVS
INEKIIEQLR AEPIPLDVIE EPVWVVKGTW KNYGV
