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New separated bacterial strain of mortierella alpine and application thereof

A technology of Mortierella alpine, strains, applied in fungi, microorganism-based methods, microorganisms, etc., can solve problems such as poor genetic stability, AA yield can not meet industrial production, reverse mutation and so on

Active Publication Date: 2014-05-21
WILMAR SHANGHAI BIOTECH RES & DEV CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the AA production of current Mortierella alpina strains still cannot meet the requirements of industrial production
Moreover, the strains used in many studies are mostly obtained by mutagenesis of low-yield natural strains. Although the yield has been improved, there are certain uncertainties in the process of use.
As we all know, mutagenized strains are less genetically stable than natural screening strains, and are prone to reverse mutations

Method used

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  • New separated bacterial strain of mortierella alpine and application thereof
  • New separated bacterial strain of mortierella alpine and application thereof
  • New separated bacterial strain of mortierella alpine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 strain screening

[0070] The original strain G-12 isolated from the soil on the hillside of Sailimu Lake, Xinjiang, was diluted on a PDA plate, and 20 single colonies that first grew out were picked and compared to shake flask experiments. Biomass produced (i.e. the amount of dry bacteria obtained per liter of fermentation broth: DCW, g / L), oil content (i.e. after breaking the wall to extract the oil, the weight percentage of the obtained oil in the dry biomass) and AA in the oil content, and one of the best strains was screened out, named Mortierella Alpina (Mortierella Alpina) WG-12N.

[0071] Experimental procedure :

[0072] 1) pick the Mortierella alpina G-12 on the original bacterial strain G-12 slope (PDA), prepare the spore suspension (concentration is 10 3 -10 4 / ml), serially diluted to 10 -3 -10 -4 Double the concentration for use.

[0073] 2) Take the diluted G-12 spore suspension and evenly spread it on a solid plate (see Table 3 for the co...

Embodiment 2

[0098] Embodiment 2 passage experiment

[0099] In this example, the Mortierella alpina strain CGMCC No. 6254 screened in Example 1 was subcultured, and shake flask fermentation experiments were performed respectively. Experimental results show that the bacterial strain of the present invention has very excellent passage stability.

[0100] Experimental procedure :

[0101] 1) The bacterial strain CGMCC No.6254 that is obtained by screening is carried out subculture fermentation test: prepare spore suspension (concentration is 10 3 -10 4 / ml), and inoculated into the seed medium at an inoculum size of 4% (v / v) (Table 7). A 250ml common Erlenmeyer bottle contains 50ml liquid, culture temperature is 25°C, shaker speed is 120rpm, culture time is 48 hours, and it is ready for use.

[0102] 2) Inoculate the above-mentioned cultured seed culture (the culture obtained in step 1) into the fermentation medium at an inoculum size of 10% (v / v) (see Table 4). A 250ml common Erlenme...

Embodiment 3

[0110] Embodiment 3 fermentation conditions

[0111] In this example, the Mortierella alpina strain CGMCC No.6254 of the present invention was subjected to a shake flask fermentation experiment.

[0112] Experimental procedure :

[0113] 1) pick CGMCC No.6254 slant (PDA) spores, make spore suspension (concentration is 10 3 -10 4 / ml), directly inoculated into the fermentation medium at an inoculum size of 4% (v / v) (see Table 4).

[0114] 2) Cultivate at 150 rpm for 8 days at 20°C, 22°C, 25°C and 28°C, respectively.

[0115] Three shake flask culture methods are: 30mL / 250mL ordinary Erlenmeyer flask, 45mL / 250mL ordinary Erlenmeyer flask and 45mL / 250mL baffle Erlenmeyer flask.

[0116] 3) Analyze the biomass, oil content and FAC content in the oil produced by each sample bottle as described in Example 1.

[0117] Experimental results :

[0118] Table 9 Experimental results of different culture methods

[0119]

[0120] Note: DCW(g / L), TFA(%), AA(%), TFA(g / L) and AA(g...

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Abstract

The invention relates to a new separated bacterial strain of mortierella alpine (Mortierella Alpina) and an application thereof. Particularly, the invention relates to CGMCC No.6254, offspring thereof, culture and / or mutant strain and a method and application in preparing arachidonic acid by using the bacterial strain, offspring thereof, culture and / or mutant strain. The separated bacterial strain is a natural bacterial strain, and can stably produce arachidonic acid with a yield as high as 14.32g / L.

Description

technical field [0001] The present invention relates to an isolated bacterial strain of Mortierella Alpina or its progeny, culture or mutant and a method for preparing arachidonic acid by using the strain or its progeny, culture or mutant and uses. Background technique [0002] The chemical name of Arachidonic Acid (AA or ARA for short) is 5,8,11,14-eicosatetraenoic acid, which belongs to the omega-6 series of long-chain polyunsaturated fatty acids (PUFA). The most important and most abundant 20-carbon polyunsaturated fatty acid. AA has a series of physiological activities such as esterifying cholesterol, inhibiting platelet aggregation, increasing blood vessel elasticity, reducing blood viscosity, regulating white blood cell function, and improving immunity. AA is also the immediate precursor of many eicosenoic acid derivatives with important physiological regulatory functions. The nutritional and health functions of AA are reflected in various aspects such as infant gro...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P7/64C12R1/645
Inventor 田桂尾姜翠红吴清杭常桂芳
Owner WILMAR SHANGHAI BIOTECH RES & DEV CENT
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