Culture medium for quickly screening filamentous fungi for producing polyunsaturated fatty acid and application of culture medium

An unsaturated fatty acid and culture medium technology is applied in the field of rapid screening of medium for polyunsaturated fatty acid-producing filamentous fungi, which can solve problems such as unreported, and achieve breakthroughs in time-consuming and labor-intensive, improving screening efficiency, and simplifying the screening process. Effect

Inactive Publication Date: 2017-06-13
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, most of the research on TTC in oil-producing microorganisms is to screen the high-yielding ARA strains according to the degree of coloring after the mycelia are dyed, while the research on cerulenin is mainly focused on the screening of high-

Method used

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  • Culture medium for quickly screening filamentous fungi for producing polyunsaturated fatty acid and application of culture medium
  • Culture medium for quickly screening filamentous fungi for producing polyunsaturated fatty acid and application of culture medium
  • Culture medium for quickly screening filamentous fungi for producing polyunsaturated fatty acid and application of culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Establishment of efficient screening method for polyunsaturated fatty acid-producing filamentous fungi

[0037] (1) Selection of bacterial strain: laboratory wild-type oleaginous filamentous fungus Mortierella alpina (Mortierella alpinaATCC 32222);

[0038] (2) Establishment of the screening method: the screening medium was based on PDA medium, and Mortierella alpina ATCC 32222, an oil-producing filamentous fungus already in our laboratory, was used as the test bacteria. Among them, the composition of the screening medium is: glucose 20g / L, potato extract 4g / L, agar 20g / L, streptomycin 30mg / L, cerulenin 1×10 -7 mol / L, TTC added concentration 0.5-2.0g / L. Incubate in the dark at 28°C for 3-7 days.

[0039] (3) Primary screening of filamentous fungi producing polyunsaturated fatty acids: prepare fresh Mortierella alpina ATCC32222 spore liquid, carry out plate coating after appropriate gradient dilution with 0.9% normal saline (the composition of plate culture m...

Embodiment 2

[0048] Example 2: Screening of filamentous fungi producing polyunsaturated fatty acids in the natural environment

[0049] Soil samples were collected from grasslands in the Qilian Mountain pastoral area at the junction of Qinghai-Gansu, and the surface soil was removed with a cleaning tool shovel, and about 150g of soil at a distance of 5-10cm was collected and packed in a sterilized centrifuge tube.

[0050] Same as Example 1, by dilution coating method, using the addition of 1×10 of the present invention -7 mol / L cerulenin and 0.5g / L triphenyltetrazolium chloride (TTC) culture medium to isolate oleaginous filamentous fungi, and culture them in the dark. The color (red) is dark and light, and the strains with fast growth speed, large colony diameter and red colony are selected as the target bacteria, and the target bacteria are transferred to the PDA medium plate for purification by using the mycelium edge purification method, and the purified bacteria are slanted. Store at...

Embodiment 3

[0053] Example 3: Screening of filamentous fungi producing polyunsaturated fatty acids in the natural environment

[0054] Soil samples were collected from the forest land of the Inner Mongolia Agricultural University campus, and the surface soil was removed with a cleaning tool shovel, and about 150 g of soil at a distance of 5-10 cm was collected and packed in a sterilized centrifuge tube.

[0055] Same as Example 1, by dilution coating method, using the addition of 1 × 10 -7 mol / L cerulenin and 0.5g / L triphenyltetrazolium chloride (TTC) culture medium to isolate oleaginous filamentous fungi, and culture them in the dark. The color (red) is light or dark, and the dark red target bacteria are transferred to the PDA medium plate for purification using the hyphae edge purification method, and the purified bacteria are stored on a slant at 4°C. Co-isolated and purified 5 strains of filamentous fungi.

[0056] The 5 strains of filamentous fungi screened were cultured in shake f...

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Abstract

The invention relates to a method for quickly screening filamentous fungi for producing polyunsaturated fatty acid. The method is characterized in that a fatty acid synthase inhibitor cerulenin is used as a resistance screening factor, triphenyltetrazolium chloride characterizing the activity of dehydrogenase is used as a screening indicator, and a method for quickly and efficiently screening the filamentous fungi for producing the polyunsaturated fatty acid at high yield from environment is established. The method disclosed by the invention is simple and quick in process; the high-yield filamentous fungi is efficiently screened, and great convenience is provided for screening and researching filamentous fungi for producing oil; meanwhile, the invention provides a high-efficiency screening method and a batch of effective original strains for producing the polyunsaturated fatty acid by utilizing microbial fermentation.

Description

[0001] 【Technical field】 [0002] The invention relates to a culture medium for rapidly screening polyunsaturated fatty acid-producing filamentous fungi and a method for rapidly screening oil-producing filamentous fungi using the medium. [0003] 【Background technique】 [0004] Polyunsaturated fatty acids (PUFAs) refer to straight-chain fatty acids containing two or more double bonds and a carbon chain length of 18-22 carbon atoms, which have a wide range of physiological functions and biological effects in the body . PUFAs are extremely important precursors in the synthesis of biomolecular eicosanoids, prostaglandins, thrombin, leukotrienes, etc. PUFAs are also important components of biological structures such as cell membrane phospholipids, and participate in the regulation of physiological metabolic processes related to biomembranes, such as cell recognition and immune response, low temperature adaptability, etc., which are extremely important for human health and the prev...

Claims

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Application Information

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IPC IPC(8): C12N1/02C12N1/14C12N1/16C12P7/64C12Q1/04G01N30/02G01N30/72
CPCC12N1/02C12N1/14C12N1/16C12P7/6427C12Q1/045G01N30/02G01N30/72
Inventor 陈海琴陈永泉陈卫姚青蔚唐鑫杨芹顾震南张灏
Owner JIANGNAN UNIV
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