SPNQ_SACSN
ID SPNQ_SACSN Reviewed; 462 AA.
AC Q9ALN8;
DT 23-MAY-2018, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-2001, sequence version 1.
DT 03-AUG-2022, entry version 64.
DE RecName: Full=dTDP-4-dehydro-2,6-dideoxy-D-glucose 3-dehydratase {ECO:0000303|PubMed:18345667};
DE EC=4.2.1.164 {ECO:0000269|PubMed:18345667};
DE AltName: Full=Pyridoxamine 5'-monophosphate-dependent 3-dehydrase {ECO:0000303|PubMed:18345667};
GN Name=spnQ {ECO:0000303|PubMed:11358695};
OS Saccharopolyspora spinosa.
OC Bacteria; Actinobacteria; Pseudonocardiales; Pseudonocardiaceae;
OC Saccharopolyspora.
OX NCBI_TaxID=60894;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND FUNCTION.
RX PubMed=11358695; DOI=10.1016/s1074-5521(01)00029-1;
RA Waldron C., Matsushima P., Rosteck P.R. Jr., Broughton M.C., Turner J.,
RA Madduri K., Crawford K.P., Merlo D.J., Baltz R.H.;
RT "Cloning and analysis of the spinosad biosynthetic gene cluster of
RT Saccharopolyspora spinosa.";
RL Chem. Biol. 8:487-499(2001).
RN [2]
RP PROTEIN SEQUENCE OF N-TERMINUS, FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, SUBUNIT, SUBSTRATE SPECIFICITY,
RP AND REACTION MECHANISM.
RC STRAIN=NRRL 18537;
RX PubMed=18345667; DOI=10.1021/ja0771383;
RA Hong L., Zhao Z., Melancon C.E. III, Zhang H., Liu H.W.;
RT "In vitro characterization of the enzymes involved in TDP-D-forosamine
RT biosynthesis in the spinosyn pathway of Saccharopolyspora spinosa.";
RL J. Am. Chem. Soc. 130:4954-4967(2008).
CC -!- FUNCTION: Involved in the biosynthesis of forosamine ((4-
CC dimethylamino)-2,3,4,6-tetradeoxy-alpha-D-threo-hexopyranose), a highly
CC deoxygenated sugar component of several bioactive natural products such
CC as the insecticidal spinosyns A and D (PubMed:11358695,
CC PubMed:18345667). Catalyzes C-3 deoxygenation of dTDP-4-keto-2,6-
CC dideoxy-alpha-D-glucose to yield dTDP-4-keto-2,3,6-trideoxy-D-glucose
CC via a combined transamination-deoxygenation reaction (PubMed:18345667).
CC The catalysis is initiated by a transamination step in which pyridoxal
CC 5'-phosphate (PLP) is converted to pyridoxamine 5'-phosphate (PMP) in
CC the presence of L-glutamate (PubMed:18345667). This coenzyme then forms
CC a Schiff base with dTDP-4-keto-2,6-dideoxy-alpha-D-glucose and the
CC resulting adduct undergoes a PMP-mediated beta-dehydration reaction to
CC give a sugar enamine intermediate, which after a 2 electrons reduction
CC and hydrolysis yields dTDP-4-keto-2,3,6-trideoxy-D-glucose as a product
CC (PubMed:18345667). Requires cellular reductase (ferredoxin or
CC flavodoxin reductase) rather than a specific partner reductase
CC (PubMed:18345667). L-glutamate is 20-fold more efficient than L-
CC aspartate as an amino donor (PubMed:18345667). In the absence of an
CC electron source and in the presence of L-glutamate, catalyzes a
CC transamination reaction, converting dTDP-4-keto-2,6-dideoxy-alpha-D-
CC glucose to dTDP-4-amino-2,4,6-trideoxy-D-glucose (PubMed:18345667).
CC {ECO:0000269|PubMed:11358695, ECO:0000269|PubMed:18345667}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=dTDP-4-dehydro-2,6-dideoxy-alpha-D-glucose + 2 H(+) + 2
CC reduced [2Fe-2S]-[ferredoxin] = dTDP-4-dehydro-2,3,6-trideoxy-alpha-
CC D-hexopyranose + H2O + 2 oxidized [2Fe-2S]-[ferredoxin];
CC Xref=Rhea:RHEA:49140, Rhea:RHEA-COMP:10000, Rhea:RHEA-COMP:10001,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:33737,
CC ChEBI:CHEBI:33738, ChEBI:CHEBI:84537, ChEBI:CHEBI:90944;
CC EC=4.2.1.164; Evidence={ECO:0000269|PubMed:18345667};
CC -!- COFACTOR:
CC Name=pyridoxal 5'-phosphate; Xref=ChEBI:CHEBI:597326;
CC Evidence={ECO:0000305|PubMed:18345667};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=49 uM for dTDP-4-keto-2,6-dideoxy-alpha-D-glucose
CC {ECO:0000269|PubMed:18345667};
CC Note=kcat is 2.6 min(-1) for dTDP-4-keto-2,6-dideoxy-alpha-D-glucose
CC as substrate. {ECO:0000269|PubMed:18345667};
CC pH dependence:
CC Optimum pH is 7.5. {ECO:0000269|PubMed:18345667};
CC Temperature dependence:
CC Optimum temperature is 37 degrees Celsius.
CC {ECO:0000269|PubMed:18345667};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:18345667}.
CC -!- SIMILARITY: Belongs to the DegT/DnrJ/EryC1 family. {ECO:0000305}.
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DR EMBL; AY007564; AAG23278.1; -; Genomic_DNA.
DR AlphaFoldDB; Q9ALN8; -.
DR SMR; Q9ALN8; -.
DR STRING; 994479.GL877879_gene4837; -.
DR KEGG; ag:AAG23278; -.
DR BioCyc; MetaCyc:MON-16621; -.
DR BRENDA; 4.2.1.164; 13744.
DR GO; GO:0016829; F:lyase activity; IEA:UniProtKB-KW.
DR CDD; cd00616; AHBA_syn; 1.
DR Gene3D; 3.40.640.10; -; 1.
DR Gene3D; 3.90.1150.10; -; 1.
DR InterPro; IPR000653; DegT/StrS_aminotransferase.
DR InterPro; IPR015424; PyrdxlP-dep_Trfase.
DR InterPro; IPR015421; PyrdxlP-dep_Trfase_major.
DR InterPro; IPR015422; PyrdxlP-dep_Trfase_small.
DR PANTHER; PTHR30244; PTHR30244; 1.
DR Pfam; PF01041; DegT_DnrJ_EryC1; 1.
DR SUPFAM; SSF53383; SSF53383; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; Lyase; Pyridoxal phosphate.
FT CHAIN 1..462
FT /note="dTDP-4-dehydro-2,6-dideoxy-D-glucose 3-dehydratase"
FT /id="PRO_0000444370"
FT ACT_SITE 246
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000250|UniProtKB:D3QY10"
FT BINDING 112..113
FT /ligand="pyridoxal 5'-phosphate"
FT /ligand_id="ChEBI:CHEBI:597326"
FT /ligand_note="ligand shared between dimeric partners"
FT /note="in other chain"
FT /evidence="ECO:0000250|UniProtKB:D3QY10"
FT BINDING 220
FT /ligand="pyridoxal 5'-phosphate"
FT /ligand_id="ChEBI:CHEBI:597326"
FT /ligand_note="ligand shared between dimeric partners"
FT /note="in other chain"
FT /evidence="ECO:0000250|UniProtKB:D3QY10"
FT BINDING 241
FT /ligand="pyridoxal 5'-phosphate"
FT /ligand_id="ChEBI:CHEBI:597326"
FT /ligand_note="ligand shared between dimeric partners"
FT /note="in other chain"
FT /evidence="ECO:0000250|UniProtKB:D3QY10"
FT BINDING 314
FT /ligand="pyridoxal 5'-phosphate"
FT /ligand_id="ChEBI:CHEBI:597326"
FT /ligand_note="ligand shared between dimeric partners"
FT /evidence="ECO:0000250|UniProtKB:D3QY10"
SQ SEQUENCE 462 AA; 50385 MW; F47895B7D805AF88 CRC64;
MQSRKTRALG KGRARVTSCD DTCATATEMV PDAKDRILAS VRDYHREQES PTFVAGSTPI
RPSGAVLDED DRVALVEAAL ELRIAAGGNA RRFESEFARF FGLRKAHLVN SGSSANLLAL
SSLTSPKLGE ARLRPGDEVI TAAVGFPTTI NPAVQNGLVP VFVDVELGTY NATPDRIKAA
VTERTRAIML AHTLGNPFAA DEIAEIAKEH ELFLVEDNCD AVGSTYRGRL TGTFGDLTTV
SFYPAHHITS GEGGCVLTGS LELARIIESL RDWGRDCWCE PGVDNTCRKR FDYHLGTLPP
GYDHKYTFSH VGYNLKTTDL QAALALSQLS KISAFGSARR RNWRRLREGL SGLPGLLLPV
ATPHSDPSWF GFAITISADA GFTRAALVNF LESRNIGTRL LFGGNITRHP AFEQVRYRIA
DALTNSDIVT DRTFWVGVYP GITDQMIDYV VESIAEFVAK SS
