ID NANA_LACPL Reviewed; 292 AA. AC P59407; F9ULC3; DT 25-MAR-2003, integrated into UniProtKB/Swiss-Prot. DT 25-MAR-2003, sequence version 1. DT 07-JUN-2017, entry version 93. DE RecName: Full=N-acetylneuraminate lyase {ECO:0000255|HAMAP-Rule:MF_01237, ECO:0000303|PubMed:21317263}; DE Short=NAL {ECO:0000255|HAMAP-Rule:MF_01237, ECO:0000303|PubMed:21317263}; DE Short=Neu5Ac lyase {ECO:0000255|HAMAP-Rule:MF_01237, ECO:0000303|PubMed:21317263}; DE EC=4.1.3.3 {ECO:0000255|HAMAP-Rule:MF_01237, ECO:0000269|PubMed:21317263}; DE AltName: Full=N-acetylneuraminate pyruvate-lyase {ECO:0000255|HAMAP-Rule:MF_01237}; DE AltName: Full=N-acetylneuraminic acid aldolase {ECO:0000255|HAMAP-Rule:MF_01237}; DE AltName: Full=Sialate lyase {ECO:0000255|HAMAP-Rule:MF_01237}; DE AltName: Full=Sialic acid aldolase {ECO:0000255|HAMAP-Rule:MF_01237, ECO:0000303|PubMed:21317263}; DE AltName: Full=Sialic acid lyase {ECO:0000255|HAMAP-Rule:MF_01237}; GN Name=nanA {ECO:0000255|HAMAP-Rule:MF_01237, GN ECO:0000303|PubMed:21317263}; OrderedLocusNames=lp_3568; OS Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1). OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Lactobacillaceae; OC Lactobacillus. OX NCBI_TaxID=220668; RN [1] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=ATCC BAA-793 / NCIMB 8826 / WCFS1; RX PubMed=12566566; DOI=10.1073/pnas.0337704100; RA Kleerebezem M., Boekhorst J., van Kranenburg R., Molenaar D., RA Kuipers O.P., Leer R., Tarchini R., Peters S.A., Sandbrink H.M., RA Fiers M.W.E.J., Stiekema W., Klein Lankhorst R.M., Bron P.A., RA Hoffer S.M., Nierop Groot M.N., Kerkhoven R., De Vries M., Ursing B., RA De Vos W.M., Siezen R.J.; RT "Complete genome sequence of Lactobacillus plantarum WCFS1."; RL Proc. Natl. Acad. Sci. U.S.A. 100:1990-1995(2003). RN [2] RP FUNCTION, CATALYTIC ACTIVITY, SUBUNIT, BIOPHYSICOCHEMICAL PROPERTIES, RP BIOTECHNOLOGY, AND 3D-STRUCTURE MODELING. RC STRAIN=ATCC BAA-793 / NCIMB 8826 / WCFS1; RX PubMed=21317263; DOI=10.1128/AEM.02927-10; RA Sanchez-Carron G., Garcia-Garcia M.I., Lopez-Rodriguez A.B., RA Jimenez-Garcia S., Sola-Carvajal A., Garcia-Carmona F., RA Sanchez-Ferrer A.; RT "Molecular characterization of a novel N-acetylneuraminate lyase from RT Lactobacillus plantarum WCFS1."; RL Appl. Environ. Microbiol. 77:2471-2478(2011). CC -!- FUNCTION: Catalyzes the reversible aldol cleavage of N- CC acetylneuraminic acid (sialic acid; Neu5Ac) to form pyruvate and CC N-acetylmannosamine (ManNAc) via a Schiff base intermediate. CC {ECO:0000255|HAMAP-Rule:MF_01237, ECO:0000269|PubMed:21317263}. CC -!- CATALYTIC ACTIVITY: N-acetylneuraminate = N-acetyl-D-mannosamine + CC pyruvate. {ECO:0000255|HAMAP-Rule:MF_01237, CC ECO:0000269|PubMed:21317263}. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=1.8 mM for N-acetylneuraminate {ECO:0000269|PubMed:21317263}; CC KM=160 mM for N-acetyl-D-mannosamine CC {ECO:0000269|PubMed:21317263}; CC KM=19.9 mM for pyruvate {ECO:0000269|PubMed:21317263}; CC Note=kcat is 10.08 sec(-1) for the cleavage of Neu5Ac and 4.8 CC sec(-1) for the synthetic reaction. CC {ECO:0000269|PubMed:21317263}; CC pH dependence: CC Optimum pH is 7-7.3 in both synthetic and cleavage directions. CC Is active over a broad pH range, from pH 5.0 to 9.0, in both CC directions. Maintains 5 to 10% activity in the synthetic CC direction above pH 11.0. The enzyme is also stable at basic pHs, CC where it maintains around 80% residual synthetic activity after CC 15 days of incubation. {ECO:0000269|PubMed:21317263}; CC Temperature dependence: CC Optimum temperature is 60 degrees Celsius for synthetic activity CC and up to 70 degrees Celsius for the cleavage reaction. Is very CC thermostable, maintaining 80% of cleavage activity after 48 CC hours at 60 degrees Celsius. However, at higher temperatures (70 CC degrees Celsius), activity decreases to less than 10% in 8 CC hours. Temperature stability is further improved by the presence CC of stabilizing additives. {ECO:0000269|PubMed:21317263}; CC -!- PATHWAY: Amino-sugar metabolism; N-acetylneuraminate degradation; CC D-fructose 6-phosphate from N-acetylneuraminate: step 1/5. CC {ECO:0000255|HAMAP-Rule:MF_01237}. CC -!- SUBUNIT: Homotetramer. {ECO:0000255|HAMAP-Rule:MF_01237, CC ECO:0000269|PubMed:21317263}. CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_01237}. CC -!- BIOTECHNOLOGY: The good synthetic activity of this enzyme at basic CC pHs, together with its thermostability and pH stability at such CC pHs, underlines the potential biotechnological application of this CC new NAL for the chemo-enzymatic synthesis of sialic acid and its CC derivatives. Thus, these characteristics make this enzyme a CC promising biocatalyst. {ECO:0000303|PubMed:21317263}. CC -!- SIMILARITY: Belongs to the DapA family. NanA subfamily. CC {ECO:0000255|HAMAP-Rule:MF_01237}. 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; AL935263; CCC80530.1; -; Genomic_DNA. DR RefSeq; WP_003646215.1; NC_004567.2. DR RefSeq; YP_004891044.1; NC_004567.2. DR ProteinModelPortal; P59407; -. DR SMR; P59407; -. DR STRING; 220668.lp_3568; -. DR EnsemblBacteria; CCC80530; CCC80530; lp_3568. DR GeneID; 1061995; -. DR KEGG; lpl:lp_3568; -. DR PATRIC; fig|220668.9.peg.2977; -. DR eggNOG; ENOG4107SCR; Bacteria. DR eggNOG; COG0329; LUCA. DR HOGENOM; HOG000173608; -. DR KO; K01639; -. DR OMA; LYEYNQC; -. DR BioCyc; LPLA220668:G137Z-2997-MONOMER; -. DR UniPathway; UPA00629; UER00680. DR Proteomes; UP000000432; Chromosome. DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell. DR GO; GO:0008747; F:N-acetylneuraminate lyase activity; IEA:UniProtKB-EC. DR GO; GO:0005975; P:carbohydrate metabolic process; IEA:UniProtKB-KW. 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 PANTHER; PTHR42849:SF1; PTHR42849:SF1; 1. DR Pfam; PF00701; DHDPS; 1. DR PIRSF; PIRSF001365; DHDPS; 1. DR PRINTS; PR00146; DHPICSNTHASE. DR SMART; SM01130; DHDPS; 1. DR PROSITE; PS00665; DHDPS_1; 1. DR PROSITE; PS00666; DHDPS_2; 1. PE 1: Evidence at protein level; KW Carbohydrate metabolism; Complete proteome; Cytoplasm; Lyase; KW Reference proteome; Schiff base. FT CHAIN 1 292 N-acetylneuraminate lyase. FT /FTId=PRO_0000103215. FT REGION 46 47 Substrate binding. {ECO:0000255|HAMAP- FT Rule:MF_01237}. FT ACT_SITE 163 163 Schiff-base intermediate with substrate. FT {ECO:0000255|HAMAP-Rule:MF_01237}. FT SITE 135 135 Involved in proton transfer during FT cleavage. {ECO:0000255|HAMAP- FT Rule:MF_01237}. SQ SEQUENCE 292 AA; 32708 MW; 4184967AE9C91747 CRC64; MSKKLLYAAQ MTAFDKDGNI NLDGIRALVR YNIDVNKVDG LYVCGSTGEA FMLNTDEKKQ VMETVYDEAN GAIDLVAQVG SLNLKEAKEL AKFATDLGYP KLSAVTPFYY NFTFEQIKDY YNEILKDVDN KLLIYSIPAL TGVALTTDQF AELFENPKII GIKYTNADFY LLERVRNAFP DKLILSGFDE MLLPALALNV DGCIGSTYNL NAPRVREEMD AFEAGDIDKA RQLQNISNDM ITDLIANDIY PTLKLVMKHM GVDAGYVKKP MSHPTPEMEA GATAIYEKYF KN //