Method of using Klebsiella michiganensis to synthesize 2'-deoxy-2-aminoadenosine
An aminoadenosine, Lebsiella technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as no breakthrough progress, reduce waste liquid discharge, reduce costs, improve The effect of utilization
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Embodiment 1
[0075] Conversion reaction conditions optimization
[0076] 1. Optimization of culture conditions for enzyme production
[0077] 1.1 The optimum culture temperature of strain DUR201505001
[0078] The 20h-cultivated seed culture medium was inoculated into the enzyme-producing medium at 5% inoculum volume, and the ventilation rate was 600L / h, and the culture was stirred at 500rpm for 20h under different temperature conditions in the temperature range of 28-42°C, and the OD660 and enzyme activity of the culture medium were determined. ( figure 2 ). The results showed that the cell mass and enzyme activity of strain DUR201505001 increased with the increase of the cultivation temperature in the culture temperature range of 28-38 °C. The enzyme activity did not change significantly or decreased when the temperature was higher than 38 °C, but the cell growth began to decrease significantly. . Therefore, the optimal culture temperature of DUR201505001 strain in the enzyme produc...
Embodiment 2
[0097] 100L-scale immobilized enzymatic synthesis of 2'-deoxy-2-aminoadenosine
[0098] 1. Bacterial preparation
[0099] 1.1 Bacterial activation and enzyme production culture
[0100] Activation medium: yeast extract 5g / L, sodium chloride 10g / L, peptone 10g / L, pH 7.0
[0101] Culture conditions: 38°C, 200rpm, 12h
[0102] Enzyme production medium: yeast extract 75g / L, sodium chloride 15g / L, corn steep liquor 60mL / L, calcium chloride 0.8g / L, magnesium sulfate heptahydrate 0.6g / L, manganese sulfate monohydrate 0.8g / L, Zinc chloride 0.1g / L, pH7.0
[0103] Culture conditions: 38°C, DO≥50%, 14h
[0104] 1.2 Bacteria collection
[0105] The enzyme-producing culture solution was centrifuged (centrifugation at 5000 rpm for 10 min), then, the wet cells were washed once with a pH 7.0, 20 mmol / L potassium dihydrogen phosphate buffer, and the wet cells obtained by centrifugation were frozen at -20 °C for later use.
[0106] 2. Preparation of Immobilized Enzyme
[0107] 2.1 Enzyme...
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