Mortierella alpine mutant strain and applications thereof

A technology of Mortierella alpina and mutant strains, applied in the direction of microorganisms, fungi, biochemical equipment and methods, etc., can solve problems such as the control of the content of long-chain saturated fatty acids that have not been mentioned, and achieve low freezing point, high oil quality, Guaranteed quality effect

Active Publication Date: 2014-03-19
CABIO BIOTECH WUHAN CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It also does not mention the control of the content of long-chain saturated fatty acids

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1. The breeding method of Mortierella alpina mutant strain

[0017] (1) Commercially available Mortierella alpina was used as the starting strain.

[0018] (2) Inoculate the strains on potato dextrose agar (PDA) medium and culture them at a constant temperature of 28°C for 8 days until the spores mature.

[0019] (3) Filter the pure spore liquid through gauze or filter paper, pour the spore liquid into a sterile culture dish and irradiate it with an ultraviolet lamp. The irradiation distance of the ultraviolet lamp is 30 cm, the irradiation time is 60 seconds, and the power of the ultraviolet lamp is 30 watts. .

[0020] (4) The spore liquid after ultraviolet mutagenesis is dried with sterile air to become bacterial plaque. Aseptically move the petri dish containing bacterial plaque into a high-energy particle beam implanter, and undergo mutagenesis by high-energy ion beam implantation.

[0021] (5) The plaques were eluted with sterile water, diluted, spre...

Embodiment 2

[0028] Embodiment 2 utilizes Mortierella alpina mutant strain to produce arachidonic acid

[0029] a) Preparation of spore suspension: Inoculate commercially available Mortierella alpina and the mutant strain of Mortierella alpina with the preservation number of the present invention: CCTCC M2013419 on a potato dextrose agar (PDA) medium plate, 25-28 Cultivate at ℃ for 7 days until the spores mature, and then scrape the spores and hyphae on the potato dextrose agar (PDA) medium plate to fill with 10 ml of sterile water, and shake to obtain a spore suspension.

[0030] b) Seed shake flask culture: inoculate the spore suspension in step (1) into the seed bottle with the culture medium, the inoculum amount is 10% (volume ratio), and place it on a shaker at 25°C and 120 rpm for 36 hours, the medium is: carbon source glucose 20g / l; nitrogen source yeast powder 5g / l; pH 5.5.

[0031] c) Fermentation bottle culture: When the bacterial concentration in the seed bottle reaches 15% (vo...

Embodiment 3

[0037] Embodiment 3 utilizes Mortierella alpina mutant strain to produce arachidonic acid

[0038]a) Preparation of spore suspension: take commercially available Mortierella alpina and the Mortierella alpina used in the present invention (preservation number: CCTCC M2013419) and inoculate them on potato dextrose agar (PDA) medium plates at 25-28°C Cultivate for 8 days until the spores mature, then scrape the spores and hyphae on the potato dextrose agar (PDA) medium plate to fill with 20 ml of sterile water, and shake to obtain the spore suspension.

[0039] b) Shake flask seed culture: inoculate the spore suspension in step (1) into the seed bottle with the medium, the inoculum amount is 15% (volume ratio), and place it on a shaker at 28°C and 220 rpm for 48 hours, the medium is: carbon source sucrose 35g / l; nitrogen source yeast extract powder 12g / l; pH7.

[0040] c) Expanded seed cultivation: the final fermenter culture adopts a volume of 50 L, so the seed tank is selected...

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Abstract

The invention relates to a mortierella alpine mutant strain ZR36 and applications thereof. Mortierella alpine ZR36 is preserved in China Center for Type Culture Collection (CCTCC) on 13th, September, 2013; preservation address is Wuhan university, Wuhan, China; and preservation number is CCTCC No.M2013419. The invention also provides applications of mortierella alpine ZR36 in production of arochidonic acid oil with low long-chain saturated fatty acid content. The content of long-chain saturated fatty acids, especially the content of long-chain saturated fatty acids containing more than twenty carbon atoms, in the arochidonic acid oil prepared using mortierella alpine ZR36 CCTCC No.M2013419 is relatively low (the total content of the long-chain saturated fatty acids containing more than twenty carbon atoms is less than 15wt%), so that freezing point of the obtained arochidonic acid oil can be reduced significantly, crystallization separation of saturated fatty acids caused by solidification of the arochidonic acid oil at relatively low temperatures is impossible, and oil quality is relatively high.

Description

technical field [0001] The invention relates to a mutant strain of Mortierella alpina and its application. Background technique [0002] Arachidonic acid (ARA) is an important polyunsaturated fatty acid (Polyunsaturated fatty acids, PUFAs), as a very important structural lipid widely exists in mammalian organs, muscles and blood tissues Among them, in addition, it is also a direct derivative of many eicosanic acid derivatives such as prostaglandin E2 (PGE2), prostacyclin (PGI2), thromboxane A2 (TXA2), leukotriene B4 (LTB4) and C4 (LTC4). These biologically active substances have important regulatory effects on lipoprotein metabolism, hemorheology, blood vessel elasticity, leukocyte function and platelet activation. [0003] Arachidonic acid oil has entered the stage of industrial production. However, most of the current research only focuses on the content of arachidonic acid in oil, but ignores the content of other components in oil, especially the content of long-chain sa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P7/64C12R1/645
Inventor 汪志明余超陆姝欢江旭伍维
Owner CABIO BIOTECH WUHAN CO LTD
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