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Literature summary for 2.3.1.4 extracted from

  • Ma, W.; Liu, Y.; Lv, X.; Li, J.; Du, G.; Liu, L.
    Combinatorial pathway enzyme engineering and host engineering overcomes pyruvate overflow and enhances overproduction of N-acetylglucosamine in Bacillus subtilis (2019), Microb. Cell Fact., 18, 001 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.3.1.4

Application

Application Comment Organism
synthesis use of enzyme for synthesis of N-acetylglucosamine in Bacillus subtilis. GNA1 is evolved through error-prone PCR under pyruvate stress to enhance its catalytic activity. Urease from Bacillus paralicheniformis is expressed intracellularly to neutralize the intracellular pH. The activity of mutant CeGNA1 increases by 11.5% at pH 6.5-7.5, with the catalytic efficiency increasing by 27.5% to 1.25 per s and microM. Modulated expression of urease increases the intracellular pH from 6.0 to 6.8. The final engineered produce 25.6 g/l N-acetylglucosamine with a yield of 0.43 g N-acetylglucosamine/g glucose in a shake flask fermentation and produces 82.5 g/l N-acetylglucosamine with a yield of 0.39 g GlcNAc/g glucose by fed-batch fermentation Caenorhabditis elegans

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Caenorhabditis elegans

Protein Variants

Protein Variants Comment Organism
Q155V/C158G activity increases by 11.5% at pH 6.5-7.5, with the catalytic efficiency increasing by 27.5% to 1.25 per s and microM Caenorhabditis elegans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.122
-
 D-glucosamine 6-phosphate mutant Q155W/C158G, pH 7.5, 30°C Caenorhabditis elegans
0.139
-
 D-glucosamine 6-phosphate wild-type, pH 7.5, 30°C Caenorhabditis elegans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
38800
-
 SDS-PAGE Caenorhabditis elegans

Organism

Organism UniProt Comment Textmining
Caenorhabditis elegans Q17427
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetyl-CoA + D-glucosamine 6-phosphate
-
 Caenorhabditis elegans CoA + N-acetyl-D-glucosamine 6-phosphate
-
 ?

Subunits

Subunits Comment Organism
? x * 19400, SDS-PAGE, mutant Q155V/C158G and wild-type Caenorhabditis elegans
dimer and monomer, 2 * 19400, SDS-PAGE Caenorhabditis elegans

Synonyms

Synonyms Comment Organism
GNA1
-
 Caenorhabditis elegans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
136
-
 D-glucosamine 6-phosphate wild-type, pH 7.5, 30°C Caenorhabditis elegans
151
-
 D-glucosamine 6-phosphate mutant Q155W/C158G, pH 7.5, 30°C Caenorhabditis elegans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
980
-
 D-glucosamine 6-phosphate wild-type, pH 7.5, 30°C Caenorhabditis elegans
1230
-
 D-glucosamine 6-phosphate mutant Q155W/C158G, pH 7.5, 30°C Caenorhabditis elegans