Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary 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

Application

EC Number Application Comment Organism
4.1.3.3 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
5.1.3.8 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)

EC Number Cloned (Comment) Organism
4.1.3.3 expressed in Escherichia coli BL21(DE3) cells Escherichia coli
4.1.3.3 expression in Escherichia coli Sus scrofa
5.1.3.8 expression in Escherichia coli Sus scrofa

General Stability

EC Number General Stability Organism
4.1.3.3 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)

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

Organism

EC Number Organism UniProt Comment Textmining
4.1.3.3 Escherichia coli
-
-
-
4.1.3.3 Escherichia coli TG1
-
-
-
4.1.3.3 Sus scrofa Q9BEC7
-
-
5.1.3.8 Sus scrofa P17560
-
-

Purification (Commentary)

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

Source Tissue

EC Number Source Tissue Comment Organism Textmining
4.1.3.3 kidney
-
Sus scrofa
-
5.1.3.8 kidney
-
Sus scrofa
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.1.3.3 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
4.1.3.3 0.0693
-
immobilized recombinant enzyme, the activity yield of immobilized enzyme is approximately 10% of the free enzyme Sus scrofa
4.1.3.3 2.45
-
crude extract, in 50 mM potassium phosphate buffer, pH 7.5, at 37°C Escherichia coli
4.1.3.3 36.26
-
after 14.8fold purification, in 50 mM potassium phosphate buffer, pH 7.5, at 37°C Escherichia coli
4.1.3.3 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
5.1.3.8 additional information
-
Escherichia coli cells harboring the recombinant plasmid are grown at 37°C in TB medium and induced by adding lactose to a final concentration of 1% lactose, samples are taken every 2 h, and the highest activity is 0.825 U/ml after fermentation for 22 h, activity is determined in 1 ml of reaction solution (100 mM Tris-HCl, pH 7.5, 100 mM GlcNAc, 10 mM MgCl2, and 2.5 mM ATP) Sus scrofa
5.1.3.8 0.07818
-
AGE, immobilized recombinant enzyme, the activity yield of immobilized enzyme is approximately 30% of the free enzyme, activity is determined in 1 ml of reaction solution (100 mM Tris-HCl, pH 7.5, 100 mM GlcNAc, 10 mM MgCl2, and 2.5 mM ATP) Sus scrofa
5.1.3.8 0.49
-
specific activity in crude extract after expression in Escherichia coli cells, activity is determined in 1 ml of reaction solution (100 mM Tris-HCl, pH 7.5, 100 mM GlcNAc, 10 mM MgCl2, and 2.5 mM ATP) Sus scrofa
5.1.3.8 29.13
-
recombinant enzyme after purification, activity is determined in 1 ml of reaction solution (100 mM Tris-HCl, pH 7.5, 100 mM GlcNAc, 10 mM MgCl2, and 2.5 mM ATP) Sus scrofa

Substrates and Products (Substrate)

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

Synonyms

EC Number Synonyms Comment Organism
4.1.3.3 N-acetyl-D-neuraminic acid lyase
-
Escherichia coli
4.1.3.3 N-acetyl-D-neuraminic acid lyase
-
Sus scrofa
4.1.3.3 NAL
-
Escherichia coli
4.1.3.3 NAL
-
Sus scrofa
5.1.3.8 AGE
-
Sus scrofa
5.1.3.8 N-acetyl-D-glucosamine-2-epimerase
-
Sus scrofa

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.1.3.3 37
-
assay at Sus scrofa
5.1.3.8 37
-
assay at Sus scrofa