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Literature summary for 4.1.3.3 extracted from

  • Hu, S.; Chen, J.; Yang, Z.; Shao, L.; Bai, H.; Luo, J.; Jiang, W.; Yang, Y.
    Coupled bioconversion for preparation of N-acetyl-D-neuraminic acid using immobilized N-acetyl-D-glucosamine-2-epimerase and N-acetyl-D-neuraminic acid lyase (2010), Appl. Microbiol. Biotechnol., 85, 1383-1391.
    View publication on PubMed
Search for more literature for 4.1.3.3

Application

Application Comment Organism
biotechnology coupled bioconversion for preparation of N-acetyl-D-neuraminic acid using immobilized N-acetyl-D-glucosamine-2-epimerase and N-acetyl-D-neuraminic acid lyase, a two-step enzymatic system involving immobilized both enzymes is used for the conversion of GlcNAc to NeuAc in a single reactor, optimum ratio is 3-6.25 U/ml of N-acetyl-D-glucosamine-2-epimerase and 12.5-25 U/ml of N-acetyl-D-neuraminic acid lyase Sus scrofa

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Escherichia coli
expression in Escherichia coli Sus scrofa

General Stability

General Stability Organism
Amberzyme oxirane resin-immobilized NAL can be used up to five reaction cycles without loss of activity or significant decrease of the conversion rate, the activity yield of immobilized NAL is approximately 10% of the free enzyme Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
N-acetyl-D-neuraminic acid Sus scrofa
-
 N-acetyl-D-mannosamine + pyruvate
-
 r

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-
Escherichia coli TG1
-
-
-
Sus scrofa Q9BEC7
-
-

Purification (Commentary)

Purification (Comment) Organism
by FP-based affinity chromatography Sus scrofa
FPIDA-Co affinity support column chromatography Escherichia coli

Source Tissue

Source Tissue Comment Organism Textmining
kidney
-
 Sus scrofa
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 cell density of Escherichia coli BL21 (DE3) carrying the pNA1 plasmid is 37 (OD600) after fermentation for 15 h, and the highest activity for N-acetyl-D-neuraminic acid lyase is 8.472 U/ml after fermentation for 9 h, enzyme activity is assayed in reaction solution (20 mM Neu5Ac in 50 mM potassium phosphate buffer, pH 7.5) Sus scrofa
0.0693
-
 immobilized recombinant enzyme, the activity yield of immobilized enzyme is approximately 10% of the free enzyme Sus scrofa
2.45
-
 crude extract, in 50 mM potassium phosphate buffer, pH 7.5, at 37°C Escherichia coli
36.26
-
 after 14.8fold purification, in 50 mM potassium phosphate buffer, pH 7.5, at 37°C Escherichia coli
36.26
-
 purified enzyme after expression in Escherichia coli cells, enzyme activity is assayed in reaction solution (20 mM N-acetyl-D-neuraminic acid in 50 mM potassium phosphate buffer, pH 7.5) Sus scrofa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
N-acetyl-D-neuraminic acid
-
 Sus scrofa N-acetyl-D-mannosamine + pyruvate
-
 r
N-acetylneuraminate
-
 Escherichia coli N-acetyl-D-mannosamine + pyruvate
-
 r
N-acetylneuraminate
-
 Escherichia coli TG1 N-acetyl-D-mannosamine + pyruvate
-
 r

Synonyms

Synonyms Comment Organism
N-acetyl-D-neuraminic acid lyase
-
 Escherichia coli
N-acetyl-D-neuraminic acid lyase
-
 Sus scrofa
NAL
-
 Escherichia coli
NAL
-
 Sus scrofa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Sus scrofa