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Candidatropicalis for producing ribonucleic acid with high yield and application thereof

A ribonucleic acid and Candida technology, which is applied in the field of biomedicine, can solve problems such as the inability to meet market demands, and achieve the effect of improving the ability to produce RNA

Active Publication Date: 2010-08-18
CHINA PHARM UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the maximum production capacity of domestic RNA is only maintained at about 200 tons, and only 100 tons of nucleotides can be produced, which is far from meeting market demand.

Method used

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  • Candidatropicalis for producing ribonucleic acid with high yield and application thereof
  • Candidatropicalis for producing ribonucleic acid with high yield and application thereof
  • Candidatropicalis for producing ribonucleic acid with high yield and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Screening of Candida tropicalis strains with high yield of ribonucleic acid (RNA)

[0040] 1.1. Strains

[0041] Tropical Candida (Candidatropicalis), ATCC14246, (purchased from the American Type Culture Collection), preserved in our laboratory

[0042] 1.2. Medium

[0043] (1) Potassium chloride sensitive screening medium A (%): glucose 3, beef extract 0.3, yeast extract 0.3, urea 0.5, FeSO 4 ·7H 2 O0.01, agar 2.5; potassium chloride sensitivity screening medium B: add 1.5mol / L KCl to medium A

[0044] (2) Seed medium S (%): glucose 4, (NH 4 ) 2 SO 4 1.5, KH 2 PO 4 0.9,K 2 HPO 4 0.3, MgSO 4 ·7H 2 O 0.3, FeSO 4 ·7H 2 O 0.003, yeast extract 0.3, corn steep liquor 0.3, CaCO 3 0.5

[0045] (3) Screening fermentation medium M (%): paraffin oil 1, urea 0.8, 80% phosphoric acid 0.3, KCl 0.15, MgSO 4 ·7H 2 O 0.1, FeSO 4 ·7H 2 O 0.003, CaCl 2 2H 2 O 0.01, NaCl 0.1, yeast extract 0.3, corn steep liquor 0.3

[0046] (4) Plate medium and slant medium C (%) f...

Embodiment 2

[0055] Medium for high RNA production of Candida tropicalis strain CPU-1

[0056] 1. Seed medium (%): Glucose 4, (NH4 ) 2 SO 4 1.5, KH 2 PO 4 0.9,K 2 HPO 4 0.3, MgSO 4 ·7H 2 O 0.3, FeSO 4 ·7H 2 O 0.003, yeast extract 0.3, corn steep liquor 0.3, CaCO 3 0.5. 200 rpm, 30°C for 16 hours. The fermentation culture conditions are as follows: the temperature of shake flask culture is 30° C., the stirring speed is 200 r / min, the inoculum size is 5%, and the seed age is 16 hours.

[0057] 2. Optimized fermentation medium (%): glucose 5, yeast extract 1.2, corn steep liquor 0.9, (NH 4 ) 2 SO 4 1.0, KH 2 PO 4 0.3. The fermentation culture conditions are: 200 rpm, 30° C., and culture for 16 hours.

Embodiment 3

[0059] Optimization of the fermentation medium of Candida tropicalis lipolytica with high yield of ribonucleic acid

[0060] 3.1. Medium

[0061] 3.1.1. Seed culture medium (%): see embodiment 2

[0062] 3.1.2. Shake flask fermentation medium (%): glucose 4, 80% phosphoric acid 0.3, (NH 4 ) 2 SO 4 0.5, KCl 0.15, MgSO 4 ·7H 2 O0.1, FeSO 4 ·7H 2 O 0.003, CaCl 2 2H 2 O 0.01, NaCl 0.1, yeast extract 0.3, corn steep liquor 0.3

[0063] 3.2. Optimization of culture medium

[0064] Optimization of carbon source: Transfer CPU-1 from the seed slant medium into 50ml seed liquid medium, and culture in a shake flask at 200rpm and 30°C for 16h. Inoculate 5% inoculum in 100ml shake flask fermentation medium, respectively use sorbitol, fructose, sucrose, dextrin, glucose, lactose and galactose as the only carbon source, culture in shake flask at 200rpm, 30℃ for 16h, measure RNA content (attached image 3 ); the result is that glucose is selected as the carbon source. 10 kinds o...

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Abstract

The invention relates to the field of biological medicine, in particular to a Candidatropicalis strain for producing ribonucleic acid with high yield and an application thereof. The Candida lipolytica strain of the invention is submitted to be preserved at China General Microbiological Culture Collection Center on December 30th, 2009, and the preserving number is CGMCCNo.3558. The strain of the invention not only has high content of nucleic acid but also has the characteristic that paraffin oil can be used by Candida lipolytica as a carbon source to be applied to industrialized production of nucleic acid.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a Candida tropicalis strain (Candida tropicalis) with high yield of ribonucleic acid (RNA) and the application of the strain with high yield of RNA. Background technique [0002] Ribonucleic acid (ribonucleic acid, RNA) is a high molecular compound in organisms, widely present in all animals, plant cells, and microorganisms, often combined with proteins to form nucleoproteins, which plays an important role in maintaining the normal functions of the body and is a specific physiological condition essential nutrients. Nucleic acid can be divided into ribonucleic acid (RNA) and deoxyribonucleic acid, referred to as DNA. DNA is the main material basis for storing, replicating and transmitting genetic information, while RNA plays an important role in the process of protein synthesis. Exogenous nucleic acid has certain functions in body nutrition, immune regulation, and disease prevention an...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12N15/01C12N13/00C12P19/34C12R1/74
Inventor 周长林方圆林忠邱蔚然曹静金由辛
Owner CHINA PHARM UNIV
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