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Method for rapid and efficient breeding of oil-producing strains and application thereof

An oil and fast technology, applied in the biological field, can solve the problems of cumbersome dyeing and extraction methods, achieve the effect of simple and efficient mutagenesis method, increase the probability of mutation, and reduce the workload of screening

Inactive Publication Date: 2018-10-23
CHINA THREE GORGES UNIV
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AI Technical Summary

Problems solved by technology

[0004] The ultimate goal of mutagenesis is to obtain strains with high oil production, but traditional staining and extraction methods are cumbersome

Method used

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  • Method for rapid and efficient breeding of oil-producing strains and application thereof
  • Method for rapid and efficient breeding of oil-producing strains and application thereof
  • Method for rapid and efficient breeding of oil-producing strains and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] ARTP mutagenesis

[0038] First make the seed liquid into OD 600 =1.0 bacterial suspension, keep the temperature of ARTP at 20°C, adjust the power to 120W, and the flow of helium to 10L / min. Take 10ul of the bacterial suspension and put it on the metal slide that has been burnt with alcohol for 30S, put the slide into the mutagenesis chamber, the emitter is 2nm away from the bacterial liquid, and use different times (0, 10, 20, 30, 40, 50, 60S) for mutagenesis treatment, put the mutagenized cells into 1 mL of normal saline and shake for 1 min, take 100 uL and spread it on a plate medium and culture it at 28°C for 2 days, and calculate the lethal rate.

[0039] Sesamol Screening

[0040] Prepare 62.5mg / mL sesamol mother solution, add 0, 40, 80, 120, 160, uL to 50mL plate medium respectively, the final concentration is 0, 0.05, 0.1, 0.15, 0.2mg / mL. OD 600 =1.0 of the seed bacteria suspension was taken and spread on the resistance plate containing sesamol, cultured at 28...

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Abstract

The invention provides a method for rapid and efficient breeding of oil-producing strains. Firstly, Mutagenesis of Trichosporon cutaneum is carried out by an atmospheric pressure and room temperatureplasma mutagenesis instrument. Then, the mutant Trichosporon cutaneum is inoculated on a flat medium containing sesamol. Strains with vigorous growth are selected and inoculated into 48-well liquid mediums, and high throughput screening is carried out on the cultured strains by 96-well plate Nile red fluorescence staining to obtain mutant strains with fast proliferation and high oil yield. Finally, oil composition is detected by gas chromatography. After mutagenesis and screening, a strain A1 with high oil yield is screened out. Compared with the original strain, the biomass, oil yield and oilproductivity of the strain A1 are increased by 41.6%, 120.3% and 58.8% respectively. The fatty acid composition of the strain A1 is the same as that of the original strain by gas chromatography composition analysis, including palmitic acid, oleic acid, linoleic acid, linolenic acid, etc. Combining ARTP plasma mutagenesis technology with malic enzyme inhibitor and Nile red fluorescent staining, high-throughput screening is realized and a strain with high oil yield is successfully bred.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an oil-producing bacterial strain, a rapid and efficient breeding method and an application thereof. Background technique [0002] Biodiesel is a clean and renewable energy that can directly replace fossil fuels. Biodiesel is a fatty acid methyl ester produced from vegetable oil and animal fat through methyl esterification. However, at present, the development of the market is greatly restricted due to its unstable source of raw materials and high cost. Microbial oil refers to the use of many microorganisms, such as yeast, algae, mold, etc., to produce and accumulate a large amount of oil in the body through microbial metabolism under certain growth conditions. The main component of the oil is triglyceride (TAG), which is similar to vegetable oil in composition. For example, palm oil and rapeseed oil are similar, mainly C16 and C18. The key to realizing high-yield micr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16C12N15/01C12N13/00C11B1/02C12R1/645
CPCC11B1/025C12N13/00C12N15/01C12N1/145C12R2001/645
Inventor 龚大春许鹏飞刘云郭金玲吕育财雷生娇田毅红
Owner CHINA THREE GORGES UNIV
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