ECTD_VIRSA
ID ECTD_VIRSA Reviewed; 300 AA.
AC Q2TDY4;
DT 16-APR-2014, integrated into UniProtKB/Swiss-Prot.
DT 24-JAN-2006, sequence version 1.
DT 03-AUG-2022, entry version 48.
DE RecName: Full=Ectoine dioxygenase {ECO:0000303|PubMed:17636255};
DE EC=1.14.11.55 {ECO:0000269|PubMed:17636255, ECO:0000269|PubMed:24714029};
DE AltName: Full=Ectoine hydroxylase {ECO:0000303|PubMed:17636255};
GN Name=ectD {ECO:0000303|PubMed:17636255};
OS Virgibacillus salexigens (Salibacillus salexigens).
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Virgibacillus.
OX NCBI_TaxID=61016;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-15, FUNCTION,
RP CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES,
RP INDUCTION, MASS SPECTROMETRY, AND COFACTOR.
RC STRAIN=ATCC 700290 / DSM 11483 / CCUG 52350 / JCM 30552 / C-20Mo;
RX PubMed=17636255; DOI=10.1074/jbc.m704023200;
RA Bursy J., Pierik A.J., Pica N., Bremer E.;
RT "Osmotically induced synthesis of the compatible solute hydroxyectoine is
RT mediated by an evolutionarily conserved ectoine hydroxylase.";
RL J. Biol. Chem. 282:31147-31155(2007).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) IN COMPLEX WITH IRON, AND COFACTOR.
RX PubMed=20498719; DOI=10.1371/journal.pone.0010647;
RA Reuter K., Pittelkow M., Bursy J., Heine A., Craan T., Bremer E.;
RT "Synthesis of 5-hydroxyectoine from ectoine: crystal structure of the non-
RT heme iron(II) and 2-oxoglutarate-dependent dioxygenase EctD.";
RL PLoS ONE 5:E10647-E10647(2010).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (1.78 ANGSTROMS), FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, AND SUBUNIT.
RX PubMed=24714029; DOI=10.1371/journal.pone.0093809;
RA Widderich N., Hoppner A., Pittelkow M., Heider J., Smits S.H., Bremer E.;
RT "Biochemical properties of ectoine hydroxylases from extremophiles and
RT their wider taxonomic distribution among microorganisms.";
RL PLoS ONE 9:e93809-e93809(2014).
CC -!- FUNCTION: Involved in the biosynthesis of 5-hydroxyectoine, called
CC compatible solute, which helps organisms to survive extreme osmotic
CC stress by acting as a highly soluble organic osmolyte. Catalyzes the 2-
CC oxoglutarate-dependent selective hydroxylation of L-ectoine to yield
CC (4S,5S)-5-hydroxyectoine. {ECO:0000269|PubMed:17636255,
CC ECO:0000269|PubMed:24714029}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-oxoglutarate + L-ectoine + O2 = 5-hydroxyectoine + CO2 +
CC succinate; Xref=Rhea:RHEA:45740, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16526, ChEBI:CHEBI:16810, ChEBI:CHEBI:30031,
CC ChEBI:CHEBI:58515, ChEBI:CHEBI:85413; EC=1.14.11.55;
CC Evidence={ECO:0000269|PubMed:17636255, ECO:0000269|PubMed:24714029};
CC -!- COFACTOR:
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000269|PubMed:17636255, ECO:0000269|PubMed:20498719,
CC ECO:0000269|PubMed:24714029};
CC Note=Binds 1 Fe(2+) ion. {ECO:0000269|PubMed:20498719,
CC ECO:0000269|PubMed:24714029};
CC -!- ACTIVITY REGULATION: Stimulated by FeSO(4) up to a concentration of 1
CC mM, whereas higher concentrations inhibited the ectoine hydroxylase.
CC Also inhibited by ascorbate. {ECO:0000269|PubMed:17636255}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.5 mM for ectoine (at pH 7.5 and 32 degrees Celsius)
CC {ECO:0000269|PubMed:17636255};
CC KM=4.9 mM for 2-oxoglutarate (at pH 7.5 and 32 degrees Celsius)
CC {ECO:0000269|PubMed:24714029};
CC KM=5.2 mM for 2-oxoglutarate (at pH 7.5 and 32 degrees Celsius)
CC {ECO:0000269|PubMed:17636255};
CC KM=5.9 mM for ectoine (at pH 7.5 and 32 degrees Celsius)
CC {ECO:0000269|PubMed:24714029};
CC Vmax=13.8 umol/min/mg enzyme (at pH 7.5 and 32 degrees Celsius)
CC {ECO:0000269|PubMed:17636255};
CC Vmax=6.4 umol/min/mg enzyme (at pH 7.5 and 32 degrees Celsius)
CC {ECO:0000269|PubMed:24714029};
CC Note=kcat is 7.7 sec(-1) for ectoin as substrate (at pH 7.5 and 32
CC degrees Celsius). {ECO:0000269|PubMed:24714029};
CC pH dependence:
CC Optimum pH is 7.5. {ECO:0000269|PubMed:17636255,
CC ECO:0000269|PubMed:24714029};
CC Temperature dependence:
CC Optimum temperature is 32 degrees Celsius (PubMed:17636255,
CC PubMed:24714029). Active from 5 to 50 degrees Celsius
CC (PubMed:24714029). {ECO:0000269|PubMed:17636255,
CC ECO:0000269|PubMed:24714029};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:24714029}.
CC -!- INDUCTION: Induced under high salinity stress and stationary growth
CC phase. Increased ectoine biosynthesis occurs as soon as the cells start
CC to grow in high osmolality medium, but there is no immediate 5-
CC hydroxyectoine production. Appreciable amounts of 5-hydroxyectoine are
CC made only when cell growth slowed and the culture enters in stationary
CC growth phase. {ECO:0000269|PubMed:17636255}.
CC -!- MASS SPECTROMETRY: Mass=34400; Method=Unknown;
CC Evidence={ECO:0000269|PubMed:17636255};
CC -!- SIMILARITY: Belongs to the PhyH family. EctD subfamily. {ECO:0000305}.
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DR EMBL; AY935522; AAY29689.1; -; Genomic_DNA.
DR PDB; 3EMR; X-ray; 1.85 A; A=1-300.
DR PDB; 4NMI; X-ray; 1.78 A; A=1-300.
DR PDBsum; 3EMR; -.
DR PDBsum; 4NMI; -.
DR AlphaFoldDB; Q2TDY4; -.
DR SMR; Q2TDY4; -.
DR KEGG; ag:AAY29689; -.
DR BRENDA; 1.14.11.55; 8050.
DR EvolutionaryTrace; Q2TDY4; -.
DR GO; GO:0016706; F:2-oxoglutarate-dependent dioxygenase activity; IDA:UniProtKB.
DR GO; GO:0005506; F:iron ion binding; IDA:UniProtKB.
DR GO; GO:0042400; P:ectoine catabolic process; IDA:UniProtKB.
DR InterPro; IPR012774; EctD.
DR InterPro; IPR008775; Phytyl_CoA_dOase.
DR Pfam; PF05721; PhyH; 1.
DR TIGRFAMs; TIGR02408; ectoine_ThpD; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Dioxygenase; Direct protein sequencing; Iron; Metal-binding;
KW Oxidoreductase.
FT CHAIN 1..300
FT /note="Ectoine dioxygenase"
FT /id="PRO_0000428756"
FT REGION 1..23
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT BINDING 129
FT /ligand="L-ectoine"
FT /ligand_id="ChEBI:CHEBI:58515"
FT /evidence="ECO:0000250|UniProtKB:Q1GNW5"
FT BINDING 135
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000250|UniProtKB:Q1GNW5"
FT BINDING 146
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:20498719,
FT ECO:0007744|PDB:3EMR"
FT BINDING 148
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:20498719,
FT ECO:0007744|PDB:3EMR"
FT BINDING 248
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:20498719,
FT ECO:0007744|PDB:3EMR"
FT SITE 152
FT /note="Important for ectoine stabilization"
FT /evidence="ECO:0000250|UniProtKB:Q1GNW5"
FT STRAND 8..11
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 14..17
FT /evidence="ECO:0007829|PDB:4NMI"
FT HELIX 27..32
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 33..35
FT /evidence="ECO:0007829|PDB:4NMI"
FT HELIX 37..46
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 47..51
FT /evidence="ECO:0007829|PDB:4NMI"
FT HELIX 57..73
FT /evidence="ECO:0007829|PDB:4NMI"
FT TURN 74..76
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 82..84
FT /evidence="ECO:0007829|PDB:4NMI"
FT TURN 86..88
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 91..95
FT /evidence="ECO:0007829|PDB:4NMI"
FT HELIX 97..100
FT /evidence="ECO:0007829|PDB:4NMI"
FT HELIX 102..108
FT /evidence="ECO:0007829|PDB:4NMI"
FT HELIX 111..121
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 125..133
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 143..146
FT /evidence="ECO:0007829|PDB:4NMI"
FT HELIX 148..156
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 163..171
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 175..177
FT /evidence="ECO:0007829|PDB:4NMI"
FT HELIX 186..188
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 189..192
FT /evidence="ECO:0007829|PDB:4NMI"
FT HELIX 216..226
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 239..243
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 248..250
FT /evidence="ECO:0007829|PDB:4NMI"
FT STRAND 261..268
FT /evidence="ECO:0007829|PDB:4NMI"
FT HELIX 269..271
FT /evidence="ECO:0007829|PDB:4NMI"
FT TURN 285..287
FT /evidence="ECO:0007829|PDB:4NMI"
SQ SEQUENCE 300 AA; 34406 MW; 0913B16BBCB96BE2 CRC64;
MEDLYPSRQN NQPKILKRKD PVIYTDRSKD NQAPITKEQL DSYEKNGFLQ IKNFFSEDEV
IDMQKAIFEL QDSIKDVASD KVIREPESND IRSIFHVHQD DNYFQDVAND KRILDIVRHL
LGSDVYVHQS RINYKPGFKG KEFDWHSDFE TWHVEDGMPR MRAISVSIAL SDNYSFNGPL
MLIPGSHNYF VSCVGETPDN NYKESLKKQK LGVPDEESLR ELTRIGGGIS VPTGKAGSVT
LFESNTMHGS TSNITPYPRN NLFMVYNSVK NRLVEPFSGG EKRPEYIAVR EKQPVYSAVN
