ID   ASPA_PSEAE              Reviewed;         474 AA.
AC   Q9HTD7;
DT   15-MAY-2007, integrated into UniProtKB/Swiss-Prot.
DT   01-MAR-2001, sequence version 1.
DT   02-SEP-2026, entry version 123.
DE   RecName: Full=Aspartate ammonia-lyase {ECO:0000303|PubMed:27988856};
DE            Short=Aspartase {ECO:0000303|PubMed:27988856};
DE            EC=4.3.1.1 {ECO:0000269|PubMed:27988856};
DE   AltName: Full=PA-AspA {ECO:0000303|PubMed:27988856};
GN   Name=aspA {ECO:0000303|PubMed:27988856}; OrderedLocusNames=PA5429;
OS   Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM
OS   14847 / LMG 12228 / 1C / PRS 101 / PAO1).
OC   Bacteria; Pseudomonadati; Pseudomonadota; Gammaproteobacteria;
OC   Pseudomonadales; Pseudomonadaceae; Pseudomonas.
OX   NCBI_TaxID=208964;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 /
RC   1C / PRS 101 / PAO1;
RX   PubMed=10984043; DOI=10.1038/35023079;
RA   Stover C.K., Pham X.-Q.T., Erwin A.L., Mizoguchi S.D., Warrener P.,
RA   Hickey M.J., Brinkman F.S.L., Hufnagle W.O., Kowalik D.J., Lagrou M.,
RA   Garber R.L., Goltry L., Tolentino E., Westbrock-Wadman S., Yuan Y.,
RA   Brody L.L., Coulter S.N., Folger K.R., Kas A., Larbig K., Lim R.M.,
RA   Smith K.A., Spencer D.H., Wong G.K.-S., Wu Z., Paulsen I.T., Reizer J.,
RA   Saier M.H. Jr., Hancock R.E.W., Lory S., Olson M.V.;
RT   "Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic
RT   pathogen.";
RL   Nature 406:959-964(2000).
RN   [2]
RP   FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP   PROPERTIES, AND BIOTECHNOLOGY.
RC   STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 /
RC   1C / PRS 101 / PAO1;
RX   PubMed=27988856; DOI=10.1007/s12010-016-2362-7;
RA   Patel A.T., Akhani R.C., Patel M.J., Dedania S.R., Patel D.H.;
RT   "Bioproduction of L-Aspartic Acid and Cinnamic Acid by L-Aspartate Ammonia
RT   Lyase from Pseudomonas aeruginosa PAO1.";
RL   Appl. Biochem. Biotechnol. 182:792-803(2017).
CC   -!- FUNCTION: Catalyzes the reversible conversion of L-aspartate to
CC       fumarate and ammonia (PubMed:27988856). Can also utilize L-
CC       phenylalanine to form cinnamic acid (PubMed:27988856). Exhibits the
CC       highest specific activity towards L-phenylalanine, but catalytic
CC       efficiency is 3-fold higher with L-aspartate (PubMed:27988856).
CC       {ECO:0000269|PubMed:27988856}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=L-aspartate = fumarate + NH4(+); Xref=Rhea:RHEA:16601,
CC         ChEBI:CHEBI:28938, ChEBI:CHEBI:29806, ChEBI:CHEBI:29991; EC=4.3.1.1;
CC         Evidence={ECO:0000269|PubMed:27988856};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=L-phenylalanine = (E)-cinnamate + NH4(+);
CC         Xref=Rhea:RHEA:21384, ChEBI:CHEBI:15669, ChEBI:CHEBI:28938,
CC         ChEBI:CHEBI:58095; Evidence={ECO:0000269|PubMed:27988856};
CC   -!- ACTIVITY REGULATION: Does not require any divalent metal ion for
CC       activation of catalysis, but the activity is slightly increased in the
CC       presence of Mg(2+), Mn(2+), Ca(2+) or Co(2+).
CC       {ECO:0000269|PubMed:27988856}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=0.346 uM for L-aspartate {ECO:0000269|PubMed:27988856};
CC         KM=1.719 mM for L-phenylalanine {ECO:0000269|PubMed:27988856};
CC         KM=21233 mM for fumarate {ECO:0000269|PubMed:27988856};
CC         Note=kcat is 4.908 sec(-1) with L-aspartate as substrate. kcat is
CC         8.002 sec(-1) with L-phenylalanine as substrate. kcat is 13.64
CC         sec(-1) with fumarate as substrate. {ECO:0000269|PubMed:27988856};
CC       pH dependence:
CC         Optimum pH is 8.0. {ECO:0000269|PubMed:27988856};
CC       Temperature dependence:
CC         Optimum temperature is 35 degrees Celsius (PubMed:27988856). Retains
CC         80% activity at 15 degrees Celsius (PubMed:27988856). Shows very high
CC         thermal stability, and retains about 56% of its activity after 7 days
CC         of incubation at 35 degrees Celsius (PubMed:27988856).
CC         {ECO:0000269|PubMed:27988856};
CC   -!- SUBUNIT: Homotetramer. {ECO:0000250|UniProtKB:P0AC38}.
CC   -!- BIOTECHNOLOGY: Catalytic efficiency establishes PA-AspA as an efficient
CC       enzyme for the production of the industrially important substances L-
CC       aspartate and cinnamic acid (PubMed:27988856). Catalytic efficiency can
CC       be enhanced through protein engineering and enzyme immobilization which
CC       can improve the production ratio (PubMed:27988856).
CC       {ECO:0000269|PubMed:27988856}.
CC   -!- SIMILARITY: Belongs to the class-II fumarase/aspartase family.
CC       Aspartase subfamily. {ECO:0000305}.
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DR   EMBL; AE004091; AAG08814.1; -; Genomic_DNA.
DR   PIR; C82968; C82968.
DR   RefSeq; NP_254116.1; NC_002516.2.
DR   RefSeq; WP_003096822.1; NC_002516.2.
DR   AlphaFoldDB; Q9HTD7; -.
DR   SMR; Q9HTD7; -.
DR   FunCoup; Q9HTD7; 157.
DR   STRING; 208964.PA5429; -.
DR   PaxDb; 208964-PA5429; -.
DR   GeneID; 878603; -.
DR   KEGG; pae:PA5429; -.
DR   PATRIC; fig|208964.12.peg.5690; -.
DR   PseudoCAP; PA5429; -.
DR   HOGENOM; CLU_021594_4_1_6; -.
DR   InParanoid; Q9HTD7; -.
DR   OrthoDB; 9802809at2; -.
DR   PhylomeDB; Q9HTD7; -.
DR   BioCyc; PAER208964:G1FZ6-5556-MONOMER; -.
DR   BRENDA; 4.3.1.1; 5087.
DR   Proteomes; UP000002438; Chromosome.
DR   AbasyAtlas; Q9HTD7; -.
DR   GO; GO:0005829; C:cytosol; IBA:GO_Central.
DR   GO; GO:0008797; F:aspartate ammonia-lyase activity; EXP:UniProtKB.
DR   GO; GO:0045548; F:phenylalanine ammonia-lyase activity; EXP:UniProtKB.
DR   GO; GO:0006531; P:aspartate metabolic process; IBA:GO_Central.
DR   GO; GO:0006099; P:tricarboxylic acid cycle; IEA:InterPro.
DR   CDD; cd01357; Aspartase; 1.
DR   FunFam; 1.10.40.30:FF:000002; Fumarate hydratase class II; 1.
DR   FunFam; 1.10.275.10:FF:000001; Fumarate hydratase, mitochondrial; 1.
DR   FunFam; 1.20.200.10:FF:000001; Fumarate hydratase, mitochondrial; 1.
DR   Gene3D; 1.10.40.30; Fumarase/aspartase (C-terminal domain); 1.
DR   Gene3D; 1.20.200.10; Fumarase/aspartase (Central domain); 1.
DR   Gene3D; 1.10.275.10; Fumarase/aspartase (N-terminal domain); 1.
DR   InterPro; IPR004708; ApsA.
DR   InterPro; IPR051546; Aspartate_Ammonia-Lyase.
DR   InterPro; IPR024083; Fumarase/histidase_N.
DR   InterPro; IPR018951; Fumarase_C_C.
DR   InterPro; IPR020557; Fumarate_lyase_CS.
DR   InterPro; IPR000362; Fumarate_lyase_fam.
DR   InterPro; IPR022761; Fumarate_lyase_N.
DR   InterPro; IPR008948; L-Aspartase-like.
DR   NCBIfam; TIGR00839; aspA; 1.
DR   NCBIfam; NF008909; PRK12273.1; 1.
DR   PANTHER; PTHR42696; ASPARTATE AMMONIA-LYASE; 1.
DR   PANTHER; PTHR42696:SF2; ASPARTATE AMMONIA-LYASE; 1.
DR   Pfam; PF10415; FumaraseC_C; 1.
DR   Pfam; PF00206; Lyase_1; 1.
DR   PRINTS; PR00149; FUMRATELYASE.
DR   SUPFAM; SSF48557; L-aspartase-like; 1.
DR   PROSITE; PS00163; FUMARATE_LYASES; 1.
PE   1: Evidence at protein level;
KW   Lyase; Reference proteome.
FT   CHAIN           1..474
FT                   /note="Aspartate ammonia-lyase"
FT                   /id="PRO_0000287771"
FT   REGION          322..331
FT                   /note="SS loop"
FT                   /evidence="ECO:0000250|UniProtKB:Q9LCC6"
FT   ACT_SITE        323
FT                   /note="Proton acceptor"
FT                   /evidence="ECO:0000250|UniProtKB:Q9LCC6"
FT   BINDING         105
FT                   /ligand="L-aspartate"
FT                   /ligand_id="ChEBI:CHEBI:29991"
FT                   /evidence="ECO:0000250|UniProtKB:Q9LCC6"
FT   BINDING         144
FT                   /ligand="L-aspartate"
FT                   /ligand_id="ChEBI:CHEBI:29991"
FT                   /evidence="ECO:0000250|UniProtKB:Q9LCC6"
FT   BINDING         145
FT                   /ligand="L-aspartate"
FT                   /ligand_id="ChEBI:CHEBI:29991"
FT                   /evidence="ECO:0000250|UniProtKB:Q9LCC6"
FT   BINDING         146
FT                   /ligand="L-aspartate"
FT                   /ligand_id="ChEBI:CHEBI:29991"
FT                   /evidence="ECO:0000250|UniProtKB:Q9LCC6"
FT   BINDING         191
FT                   /ligand="L-aspartate"
FT                   /ligand_id="ChEBI:CHEBI:29991"
FT                   /evidence="ECO:0000250|UniProtKB:Q9LCC6"
FT   BINDING         324
FT                   /ligand="L-aspartate"
FT                   /ligand_id="ChEBI:CHEBI:29991"
FT                   /evidence="ECO:0000250|UniProtKB:Q9LCC6"
FT   BINDING         329
FT                   /ligand="L-aspartate"
FT                   /ligand_id="ChEBI:CHEBI:29991"
FT                   /evidence="ECO:0000250|UniProtKB:Q9LCC6"
SQ   SEQUENCE   474 AA;  51070 MW;  DE868A8CA8478429 CRC64;
     MSPVASSRIE KDLLGTLEVP ADAYYGIQTL RAVNNFRLSG VPLSHYPKLV VALAMVKQAA
     ADANRQLGHL PEDKHAAISE ACARLIRGDF HEQFVVDMIQ GGAGTSTNMN ANEVIANIAL
     EAMGHTKGEY KYLHPNNDVN MAQSTNDAYP TAIRLGLLLG HDTLLASLDS LIQAFAAKGV
     EFAGVLKMGR TQLQDAVPMT LGQEFHAFAT TLGEDLDRLR RLAPELLTEV NLGGTAIGTG
     INADPGYQKL AVERLAAISG QPLKPAADLI EATSDMGAFV LFSGMLKRTA VKLSKICNDL
     RLLSSGPRTG INEINLPPRQ PGSSIMPGKV NPVIPEAVNQ VAFEVIGNDL ALTLAAEGGQ
     LQLNVMEPLI AYKIFDSIRL LQRAMDMLRE HCITGITANV ERCHELVEHS IGLVTALNPY
     IGYENSTRIA KTALESGRGV LELVREEKLL DEATLADILL PENMIAPRLI PLRA
//