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A method for synthesizing uracil by Pseudomonas aeruginosa

A technology of Pseudomonas aeruginosa and uracil, which is applied in the field of biological fermentation in biomedicine, can solve problems such as no breakthrough progress, and achieve the effects of environmental protection, simple extraction process and cost reduction.

Active Publication Date: 2019-01-15
TUOXIN GROUP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its synthesis method is mainly chemical synthesis method, and there is still no breakthrough in the direct synthesis of uracil from cytosine by biosynthesis

Method used

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  • A method for synthesizing uracil by Pseudomonas aeruginosa
  • A method for synthesizing uracil by Pseudomonas aeruginosa
  • A method for synthesizing uracil by Pseudomonas aeruginosa

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Conversion reaction condition optimization

[0045] 1. Optimization of culture conditions for enzyme production

[0046] 1.1 The optimal culture temperature of CUR201604001 strain

[0047] Inoculate the seed culture solution cultivated for 12 hours in the enzyme-producing medium according to the inoculum amount of 5%, and incubate at 500 rpm under different temperature conditions in the temperature range of 28-42°C for 16 hours with a ventilation rate of 600L / h, and measure the OD660 and enzyme activity of the culture solution ( figure 2 ). The results showed that the cell mass and enzyme activity of CUR2016 04001 strain increased with the increase of culture temperature in the range of 28-36°C, and the enzyme activity and cell mass decreased significantly above 38°C. Therefore, the optimal temperature range for enzyme production of CUR201604001 strain was 36-38°C.

[0048] 1.2 Optimum pH value for culture of CUR201604001 strain

[0049] Inoculate the seed culture...

Embodiment 2

[0065] Scale-up of 100L-scale synthesis of uracil using Pseudomonas aeruginosa

[0066] 1. Cell preparation

[0067] 1.1 Cell activation and enzyme production culture

[0068] Activation medium: yeast extract 5g / L, sodium chloride 10g / L, peptone 10g / L, pH7.0;

[0069] Culture conditions: 35°C, 200rpm, 24h;

[0070] 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;

[0071] Culture conditions: 38°C, DO≥50%, 20h.

[0072] 1.2 Cell collection

[0073] The enzyme-producing culture solution was concentrated by microfiltration, and then the wet bacteria were collected by centrifugation, and stored at -20°C until use.

[0074] 2. Enzymatic reaction

[0075] Reaction system preparation: Cytosine 40Kg; wet bacteria volume: 12Kg; purified water 100L; pH: 7.0. After the preparation of the reaction system is...

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Abstract

The invention discloses a method for synthesizing uracil by using Pseudomonas aeruginosa, belonging to the technical field of biological fermentation. Cytosine is converted into uracil in one step byusing Pseudomonas aeruginosa as bacterial strain and wet cell as enzyme source. Compared with the literature, the reaction rate and conversion rate of uracil produced by this process are faster, the conversion rate of substrate is more than 99%, the process is simple, the cost is low, and the operation is suitable for industrialization.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation in biomedicine, and relates to the biosynthesis of pyrimidine bases, in particular to a method for synthesizing uracil by using Pseudomonas aeruginosa. Background technique [0002] Pseudomonas aeruginosa (Pseudomonas aeruginosa) was formerly known as Pseudomonas aeruginosa. It is widely distributed in nature and is one of the most common bacteria in soil. All kinds of water, air, skin, respiratory tract and intestinal tract of normal people have this bacteria. The important condition for the existence of this bacterium is a humid environment. This bacterium is a common opportunistic pathogen and belongs to non-fermenting Gram-negative bacilli. The bacteria are slender and of different lengths, sometimes club-shaped or linear, arranged in pairs or short chains. There is a single flagella at one end of the bacterium, and the bacterial movement can be seen under a dark-field micr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/12C12N1/20C12R1/385
CPCC12P17/12C12N1/205C12R2001/385
Inventor 杨西宁邢善涛李涛王东琨卓凯悦柳芳
Owner TUOXIN GROUP