P59407 · NANA_LACPL
- ProteinN-acetylneuraminate lyase
- GenenanA
- StatusUniProtKB reviewed (Swiss-Prot)
- Amino acids292 (go to sequence)
- Protein existenceEvidence at protein level
- Annotation score4/5
Function
function
Catalyzes the reversible aldol cleavage of N-acetylneuraminic acid (sialic acid; Neu5Ac) to form pyruvate and N-acetylmannosamine (ManNAc) via a Schiff base intermediate.
Catalytic activity
- aceneuramate = aldehydo-N-acetyl-D-mannosamine + pyruvate
Biotechnology
The good synthetic activity of this enzyme at basic pHs, together with its thermostability and pH stability at such pHs, underlines the potential biotechnological application of this new NAL for the chemo-enzymatic synthesis of sialic acid and its derivatives. Thus, these characteristics make this enzyme a promising biocatalyst.
Kinetics
KM | SUBSTRATE | pH | TEMPERATURE[C] | NOTES | EVIDENCE | |
---|---|---|---|---|---|---|
1.8 mM | N-acetylneuraminate | |||||
160 mM | N-acetyl-D-mannosamine | |||||
19.9 mM | pyruvate |
kcat is 10.08 sec-1 for the cleavage of Neu5Ac and 4.8 sec-1 for the synthetic reaction.
pH Dependence
Optimum pH is 7-7.3 in both synthetic and cleavage directions. Is active over a broad pH range, from pH 5.0 to 9.0, in both directions. Maintains 5 to 10% activity in the synthetic direction above pH 11.0. The enzyme is also stable at basic pHs, where it maintains around 80% residual synthetic activity after 15 days of incubation.
Temperature Dependence
Optimum temperature is 60 degrees Celsius for synthetic activity and up to 70 degrees Celsius for the cleavage reaction. Is very thermostable, maintaining 80% of cleavage activity after 48 hours at 60 degrees Celsius. However, at higher temperatures (70 degrees Celsius), activity decreases to less than 10% in 8 hours. Temperature stability is further improved by the presence of stabilizing additives.
Pathway
Amino-sugar metabolism; N-acetylneuraminate degradation; D-fructose 6-phosphate from N-acetylneuraminate: step 1/5.
Features
Showing features for binding site, active site.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Binding site | 46 | aceneuramate (UniProtKB | ChEBI) | ||||
Sequence: S | ||||||
Binding site | 47 | aceneuramate (UniProtKB | ChEBI) | ||||
Sequence: T | ||||||
Active site | 135 | Proton donor | ||||
Sequence: Y | ||||||
Active site | 163 | Schiff-base intermediate with substrate | ||||
Sequence: K | ||||||
Binding site | 165 | aceneuramate (UniProtKB | ChEBI) | ||||
Sequence: T | ||||||
Binding site | 187 | aceneuramate (UniProtKB | ChEBI) | ||||
Sequence: G | ||||||
Binding site | 189 | aceneuramate (UniProtKB | ChEBI) | ||||
Sequence: D | ||||||
Binding site | 190 | aceneuramate (UniProtKB | ChEBI) | ||||
Sequence: E | ||||||
Binding site | 206 | aceneuramate (UniProtKB | ChEBI) | ||||
Sequence: S |
GO annotations
Aspect | Term | |
---|---|---|
Cellular Component | cytosol | |
Molecular Function | N-acetylneuraminate lyase activity | |
Biological Process | carbohydrate metabolic process | |
Biological Process | N-acetylneuraminate catabolic process |
Keywords
- Molecular function
- Biological process
- Ligand
Enzyme and pathway databases
Names & Taxonomy
Protein names
- Recommended nameN-acetylneuraminate lyase
- EC number
- Short namesNAL ; Neu5Ac lyase
- Alternative names
Gene names
Organism names
- Strain
- Taxonomic lineageBacteria > Bacillota > Bacilli > Lactobacillales > Lactobacillaceae > Lactiplantibacillus
Accessions
- Primary accessionP59407
- Secondary accessions
Proteomes
Subcellular Location
Phenotypes & Variants
PTM/Processing
Features
Showing features for chain.
Type | ID | Position(s) | Description | |||
---|---|---|---|---|---|---|
Chain | PRO_0000103215 | 1-292 | N-acetylneuraminate lyase | |||
Sequence: MSKKLLYAAQMTAFDKDGNINLDGIRALVRYNIDVNKVDGLYVCGSTGEAFMLNTDEKKQVMETVYDEANGAIDLVAQVGSLNLKEAKELAKFATDLGYPKLSAVTPFYYNFTFEQIKDYYNEILKDVDNKLLIYSIPALTGVALTTDQFAELFENPKIIGIKYTNADFYLLERVRNAFPDKLILSGFDEMLLPALALNVDGCIGSTYNLNAPRVREEMDAFEAGDIDKARQLQNISNDMITDLIANDIYPTLKLVMKHMGVDAGYVKKPMSHPTPEMEAGATAIYEKYFKN |
Interaction
Structure
Sequence
- Sequence statusComplete
- Length292
- Mass (Da)32,708
- Last updated2003-03-25 v1
- Checksum4184967AE9C91747
Keywords
- Technical term
Sequence databases
Nucleotide Sequence | Protein Sequence | Molecule Type | Status | |
---|---|---|---|---|
AL935263 EMBL· GenBank· DDBJ | CCC80530.1 EMBL· GenBank· DDBJ | Genomic DNA |