from yeast overproducer, lyophilizate
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Hexokinase (HK) material number and pack size: Material Number Pack Size 11119796103 custom fill
- Recombinant enzyme that converts hexose to hexose-6-phosphate.
- Use Hexokinase in diagnostic tests for blood glucose using the hexokinase method and for the determination of creatine kinase.
Nomenclature: ATP:D-hexose 6-phosphotransferase
Molecular weight: 57 kD (SDS-PAGE)
Isoelectric point: 4.5-5.0
Michaelis constants (D-glucose):
Phosphate buffer, 0.1 mol/L, pH 7.0; +25°C: 3.05 x 10-4 mol/L
Phosphate buffer, 0.1 mol/L, pH 7.4; +30°C: 1.90 x 10-4 mol/L
Tea buffer, 0.1 mol/L, pH 7.6; +25°C: 2.30 x 10-4 mol/L
Michaelis constants (ATP):
Tris buffer, 0.1 mol/L, pH 7.6; +28°C: 1.60 x 10-4 mol/L
Tea buffer, 0.1 mol/L, pH 7.6; +25°C: 1.90 x 10-4 mol/L
Inhibitors: EDTA, SH-blocking compounds, sorbose-1-phosphate, polyphosphates, 6-deoxy-6-fluoroglucose, 2-C-hydroxymethylglucose, lyxose.
Activators: Mg2+, catecholamines
pH optimum: 7.0-10.0 (see figure)
Temperature dependence: See figure
pH stability: 5.0-9.0 (see figure)
Thermal stability: 100% for 20 minutes at +37°C, 50% for 20 minutes at +45°C, 75% for 5 hours at +37°C (see figures)
Specificity: Hexokinase phosphorylates D-glucose, D-fructose, D-mannose, D-glucosamin, 2-deoxyglucose. L-Arabinose, D-xylose, L-rhamnose, D-galactose, D-lactose, sucrose, maltose, trehalose, raffinose, N-acetyl glucosamine do not react. ATP can be partially replaced by other nucleotides. Appearance: Yellowish lyophilizate
Solubility: Clear, colorless solution in water (c=10 mg/mL)
pH value (c=10 mg/mL in water): 6.5-7.5
Activity (+25°C, glucose): ≥70 U/mg lyophilizate
Activity (+30°C): ≥98 U/mg lyophilizate
Activity (+37°C): ≥115 U/mg
Protein (Biuret): 0.15±0.05 mg/mg lyophilizate
Contaminants (expressed as percentage of Hexokinase activity):
Alcohol dehydrogenase: ≤0.001
Creatine kinase: ≤0.001
Glutamate dehydrogenase: ≤0.05
Glutathione reductase: ≤0.005
''NADH oxidase'': ≤0.001
''NADPH oxidase'': ≤0.001
6-Phosphogluconate dehydrogenase: ≤0.001
Phophoglucose isomerase: ≤0.002
Stability: At +2 to +8°C within specification range for 18 months. Store dry.