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 (Comment) | Organism |
---|---|
expression in Escherichia coli | Caenorhabditis elegans |
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 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 Maximum [Da] | Comment | Organism |
---|---|---|---|
38800 | - |
SDS-PAGE | Caenorhabditis elegans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Caenorhabditis elegans | Q17427 | - |
- |
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 | Comment | Organism |
---|---|---|
? | x * 19400, SDS-PAGE, mutant Q155V/C158G and wild-type | Caenorhabditis elegans |
dimer | and monomer, 2 * 19400, SDS-PAGE | Caenorhabditis elegans |
Synonyms | Comment | Organism |
---|---|---|
GNA1 | - |
Caenorhabditis elegans |
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 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 |