NANA_CLOPE
ID NANA_CLOPE Reviewed; 288 AA.
AC Q9S4K9; O08360;
DT 25-MAR-2003, integrated into UniProtKB/Swiss-Prot.
DT 25-MAR-2003, sequence version 2.
DT 03-AUG-2022, entry version 125.
DE RecName: Full=N-acetylneuraminate lyase;
DE Short=NAL;
DE Short=Neu5Ac lyase;
DE EC=4.1.3.3;
DE AltName: Full=N-acetylneuraminate pyruvate-lyase;
DE AltName: Full=N-acetylneuraminic acid aldolase;
DE AltName: Full=Sialate lyase;
DE AltName: Full=Sialic acid aldolase;
DE AltName: Full=Sialic acid lyase;
GN Name=nanA; OrderedLocusNames=CPE0185;
OS Clostridium perfringens (strain 13 / Type A).
OC Bacteria; Firmicutes; Clostridia; Eubacteriales; Clostridiaceae;
OC Clostridium.
OX NCBI_TaxID=195102;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PROTEIN SEQUENCE OF 1-31.
RC STRAIN=A99;
RX PubMed=9511987; DOI=10.1023/a:1018585920853;
RA Traving C., Roggentin P., Schauer R.;
RT "Cloning, sequencing and expression of the acetylneuraminate lyase gene
RT from Clostridium perfringens A99.";
RL Glycoconj. J. 14:821-830(1997).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND CHARACTERIZATION.
RC STRAIN=NCTC 8798 / Type A;
RX PubMed=10419949; DOI=10.1128/jb.181.15.4526-4532.1999;
RA Walters D.M., Stirewalt V.L., Melville S.B.;
RT "Cloning, sequence, and transcriptional regulation of the operon encoding a
RT putative N-acetylmannosamine-6-phosphate epimerase (nanE) and sialic acid
RT lyase (nanA) in Clostridium perfringens.";
RL J. Bacteriol. 181:4526-4532(1999).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=13 / Type A;
RX PubMed=11792842; DOI=10.1073/pnas.022493799;
RA Shimizu T., Ohtani K., Hirakawa H., Ohshima K., Yamashita A., Shiba T.,
RA Ogasawara N., Hattori M., Kuhara S., Hayashi H.;
RT "Complete genome sequence of Clostridium perfringens, an anaerobic flesh-
RT eater.";
RL Proc. Natl. Acad. Sci. U.S.A. 99:996-1001(2002).
RN [4]
RP CHARACTERIZATION, AND MUTAGENESIS OF TYR-133; LYS-161; ASP-187 AND GLU-188.
RX PubMed=11432751; DOI=10.1046/j.1432-1327.2001.02297.x;
RA Krueger D., Schauer R., Traving C.;
RT "Characterization and mutagenesis of the recombinant N-acetylneuraminate
RT lyase from Clostridium perfringens: insights into the reaction mechanism.";
RL Eur. J. Biochem. 268:3831-3839(2001).
CC -!- FUNCTION: Catalyzes the reversible aldol cleavage of N-acetylneuraminic
CC acid (sialic acid; Neu5Ac) to form pyruvate and N-acetylmannosamine
CC (ManNAc) via a Schiff base intermediate.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=aceneuramate = N-acetyl-D-mannosamine + pyruvate;
CC Xref=Rhea:RHEA:23296, ChEBI:CHEBI:15361, ChEBI:CHEBI:17122,
CC ChEBI:CHEBI:173083; EC=4.1.3.3;
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=3.5 mM for Neu5Ac;
CC Vmax=27.5 umol/min/mg enzyme;
CC pH dependence:
CC Optimum pH is 7.6.;
CC Temperature dependence:
CC Optimum temperature is 65-70 degrees Celsius.;
CC -!- PATHWAY: Amino-sugar metabolism; N-acetylneuraminate degradation; D-
CC fructose 6-phosphate from N-acetylneuraminate: step 1/5.
CC -!- SUBUNIT: Homotetramer. {ECO:0000305}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm.
CC -!- INDUCTION: By N-acetylneuraminate.
CC -!- SIMILARITY: Belongs to the DapA family. NanA subfamily. {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; Y12876; CAA73375.1; -; Genomic_DNA.
DR EMBL; AF130859; AAD28763.1; -; Genomic_DNA.
DR EMBL; BA000016; BAB79891.1; -; Genomic_DNA.
DR RefSeq; WP_003457546.1; NC_003366.1.
DR AlphaFoldDB; Q9S4K9; -.
DR SMR; Q9S4K9; -.
DR STRING; 195102.gene:10489429; -.
DR EnsemblBacteria; BAB79891; BAB79891; BAB79891.
DR GeneID; 29572700; -.
DR KEGG; cpe:CPE0185; -.
DR HOGENOM; CLU_049343_6_0_9; -.
DR OMA; VVPPVCT; -.
DR BioCyc; MetaCyc:MON-18990; -.
DR UniPathway; UPA00629; UER00680.
DR Proteomes; UP000000818; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0008747; F:N-acetylneuraminate lyase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:UniProtKB-UniRule.
DR GO; GO:0019262; P:N-acetylneuraminate catabolic process; IEA:UniProtKB-UniPathway.
DR CDD; cd00954; NAL; 1.
DR Gene3D; 3.20.20.70; -; 1.
DR HAMAP; MF_01237; N_acetylneuram_lyase; 1.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR002220; DapA-like.
DR InterPro; IPR005264; NanA.
DR InterPro; IPR020625; Schiff_base-form_aldolases_AS.
DR InterPro; IPR020624; Schiff_base-form_aldolases_CS.
DR Pfam; PF00701; DHDPS; 1.
DR PIRSF; PIRSF001365; DHDPS; 1.
DR PRINTS; PR00146; DHPICSNTHASE.
DR SMART; SM01130; DHDPS; 1.
DR TIGRFAMs; TIGR00683; nanA; 1.
DR PROSITE; PS00665; DHDPS_1; 1.
DR PROSITE; PS00666; DHDPS_2; 1.
PE 1: Evidence at protein level;
KW Carbohydrate metabolism; Cytoplasm; Direct protein sequencing; Lyase;
KW Reference proteome; Schiff base.
FT CHAIN 1..288
FT /note="N-acetylneuraminate lyase"
FT /id="PRO_0000103208"
FT ACT_SITE 161
FT /note="Schiff-base intermediate with substrate"
FT BINDING 44..45
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT SITE 133
FT /note="Involved in proton transfer during cleavage"
FT VARIANT 211
FT /note="V -> I (in strain: NCTC 8798)"
FT VARIANT 278
FT /note="A -> E (in strain: NCTC 8798)"
FT MUTAGEN 133
FT /note="Y->F,H,W,C: Large decrease in activity but increase
FT in substrate affinity."
FT /evidence="ECO:0000269|PubMed:11432751"
FT MUTAGEN 161
FT /note="K->Q,A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:11432751"
FT MUTAGEN 161
FT /note="K->R: 33-fold decrease in activity but no change in
FT substrate affinity."
FT /evidence="ECO:0000269|PubMed:11432751"
FT MUTAGEN 187
FT /note="D->E: 9-fold decrease in substrate affinity but
FT almost no change in activity."
FT /evidence="ECO:0000269|PubMed:11432751"
FT MUTAGEN 187
FT /note="D->N: Large decrease in substrate affinity and
FT activity."
FT /evidence="ECO:0000269|PubMed:11432751"
FT MUTAGEN 188
FT /note="E->D,Q: Decrease in substrate affinity but almost no
FT change in activity."
FT /evidence="ECO:0000269|PubMed:11432751"
SQ SEQUENCE 288 AA; 32387 MW; C5938737636AD3DC CRC64;
MKGIYSALLV SFDKDGNINE KGLREIIRHN IDVCKIDGLY VGGSTGENFM LSTDEKKRIF
EIAMDEAKGQ VKLIAQVGSV NLKEAVELAK FTTDLGYDAI SAVTPFYYKF DFNEIKHYYE
TIINSVDNKL IIYSIPFLTG VNMSIEQFAE LFENDKIIGV KFTAADFYLL ERMRKAFPDK
LIFAGFDEMM LPATVLGVDG AIGSTFNVNG VRARQIFEAA QKGDIETALE VQHVTNDLIT
DILNNGLYQT IKLILQEQGV DAGYCRQPMK EATEEMIAKA KEINKKYF
