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Bacterial strain for producing polyunsaturated fatty acids and screening method thereof

A technology of unsaturated fatty acid and screening method, applied in microorganism-based methods, biochemical equipment and methods, fungi, etc., can solve the problems of inability to achieve industrialized production, large variation, etc., to save screening costs and grow vigorously. , to avoid random effects

Inactive Publication Date: 2014-10-15
无锡超科食品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, in the prior art, mutagenesis screening is usually used to obtain polyunsaturated fatty acid-producing strains, which has the disadvantage of large variation and the inability to realize industrial production.

Method used

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  • Bacterial strain for producing polyunsaturated fatty acids and screening method thereof
  • Bacterial strain for producing polyunsaturated fatty acids and screening method thereof
  • Bacterial strain for producing polyunsaturated fatty acids and screening method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Low temperature culture for primary screening

[0041] The nine soil samples (see Table 1) from the three regions are labeled as A1, A2, A3, B1, B2, B3, C1, C2, and C3, respectively. Weigh 1g of soil sample each, add it into a 500mL Erlenmeyer flask filled with 100mL sterile water and glass beads, shake for 15min, and take 150μL suspension (dilution equivalent to 10 -2 ) on a PDA plate (three plates for each sample), and cultured at 5°C and 28°C, respectively.

[0042] Table 1 Basic conditions of collected soil

[0043]

[0044] The culture results are shown in Table 2. Under the low temperature condition of 5°C, the cultured microorganisms grow very slowly, and it takes about 7 days to grow enough colonies, most of which are moist bacterial colonies and a few filamentous fungal colonies; After 2 days of culture, more colonies appeared on the plates. After comparison, it can be seen that the culture at low temperature at 5°C inhibits the growth of many m...

Embodiment 2

[0048] Example 2 Sudan black staining microscopic examination for re-screening

[0049] 80 colonies were selected from the plate grown at low temperature for Sudan black staining, and the oil particles in the bacteria were observed under a microscope. The specific dyeing method is as follows:

[0050] 1) Activation of bacteria: transplant 80 selected colonies to a fresh PDA medium slant for purification, isolation and numbering, and culture at 28°C for three days;

[0051] 2) Smear: Drop a small drop of distilled water in the center of a clean glass slide, use an inoculation loop to aseptically pick a little bacterial lawn from the slant culture in the water drop, mix it and paint it into a film;

[0052] 3) drying: natural drying at room temperature;

[0053] 4) Fixing: Hold one end of the glass slide with the bacteria-coated side facing up, and pass through the alcohol flame 2-3 times;

[0054] 5) Staining: place the smear in a horizontal position, add Sudan black dyeing ...

Embodiment 3

[0062] Embodiment 3 liquid fermentation culture

[0063] Seed liquid culture: Inoculate five strains of 1C12, 2A11, 2A12, 2B31, and 2A34 obtained through screening into a 250mL Erlenmeyer flask containing 50mL of seed medium, and culture them on a shaker at 30°C for 3 days at 150r / min as a fermentation medium. Use strains.

[0064] Shake flask fermentation culture: After breaking up the bacteria for fermentation with a disperser, inoculate 10% of the inoculum into a 1L Erlenmeyer flask with a baffle (the liquid volume is 200ml), at 30°C, 150r / min shaker culture 120h.

[0065] Determination of biomass: After the whole fermentation liquid was broken up by a disperser, it was filtered by suction and washed 3 times with distilled water. The bacteria were collected in a 9cm plate, freeze-dried and weighed.

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Abstract

The invention discloses a bacterial strain for producing polyunsaturated fatty acids (PUFAs) and a screening method thereof. The bacterial strain is named as Mucor circinelloides in taxonomy, and the preservation number of the bacterial strain is CGMCC No.8361. The screening method comprises the following steps: (1) prescreening at a low temperature; (2) dyeing the bacterial strain with Sudan black and rescreening; (3) fermenting and culturing the bacterial strain in liquid; and (4) using gas chromatography for detecting the content of fatty acids in bacterial strain cells. The bacterial strain obtained by the screening method disclosed by the invention can produce the polyunsaturated fatty acids in high yield. Through the adoption of the screening method disclosed by the invention, the bacterial strain can be obtained quickly and repeatedly, so that the screening method has the advantages of low cost and capability of being applied to industrialized production, and the like.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a bacterial strain and a screening method thereof, in particular to a polyunsaturated fatty acid-producing bacterial strain and a screening method thereof. Background technique [0002] Polyunsaturated fatty acids (PUFAs) refer to straight-chain fatty acids with two or more carbon-carbon double bonds and 16-22 carbon atoms. According to the position of the first double bond in the carbon chain, PUFAs can be divided into ω-3, ω-6 and ω-9 series, but the ω-3 and ω-6 PUFAs have important biological significance. ω-3PUFAs mainly include α-linolenic acid (ALA), stearidonic acid, eicosadonic acid, eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and Dicosahexaenoic acid (DHA). ω-6PUFAs mainly include linoleic acid (LA), γ-linolenic acid (GLA), double homo-γ-linolenic acid (DGLA) and arachidonic acid (AA). [0003] Studies have shown that PUFAs have many health care functions fo...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N1/02C12R1/785
Inventor 刘小鸣熊文珂蒋瑜王萍李敏江丽红
Owner 无锡超科食品有限公司
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