Mortierella alpina mutant strain, method for producing arachidonic acid oil using it, and arachidonic acid oil
A technology of Mortierella alpine and arachidonic acid, which is applied in the field of Mortierella alpine mutants, can solve the problems of reducing the content of long-chain saturated fatty acids, which are not mentioned, and achieve the effect of maintaining clarity, transparency and high quality.
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Embodiment 1
[0020] Breeding method of Mortierella alpina mutant strain
[0021] (1) Take the CCTCC NO:M2013419 strain as the starting strain, which was preserved in the China Center for Type Culture Collection (CCTCC) by Carbio Bioengineering (Wuhan) Co., Ltd. on September 13, 2013. The preservation address is, China, Wuhan, Wuhan University, deposit number is CCTCC NO:M2013419.
[0022] (2) The bacteria were inoculated on a potato dextrose agar (PDA) medium and cultured at a constant temperature of 28° C. for 8 days until the spores matured.
[0023] (3) Obtain pure spore liquid by filtering through gauze or filter paper, inject the spore liquid into a sterile culture dish and pass through ultraviolet lamp irradiation, the irradiation distance of ultraviolet lamp is 15 cm, the irradiation time is 120 seconds, and the power of ultraviolet lamp is 30 watts .
[0024] (4) The spore liquid after ultraviolet mutagenesis is air-dried with sterile air to become bacterial plaque. Aseptically ...
Embodiment 2
[0031] Production of arachidonic acid by Mortierella alpina mutant strain
[0032] a) Preparation of spore suspension: Inoculate commercially available Mortierella alpina and Mortierella alpina used in the present invention (preservation number: CCTCC M2015421) on a potato dextrose agar (PDA) medium plate, 25-27°C Cultivate for 10 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 a spore suspension.
[0033]b) Shake flask seed culture: inoculate the spore suspension of step (1) into the seed bottle with medium, inoculum size 15% (volume ratio), place 27 ° C, 220 rev / min shaker to cultivate 72 hours, the medium is: carbon source sucrose 35g / l; nitrogen source yeast extract powder 12g / l; pH 7.
[0034] c) Expanded seed cultivation: the final fermenter culture adopts a volume of 50 L, so the seed tank is selected as a 10 L seed enlarged fermenter. Inoculate the ...
Embodiment 3
[0042] Production of arachidonic acid by Mortierella alpina mutant strain
[0043] a) Preparation of spore suspension: take commercially available Mortierella alpina and the mutant strain of Mortierella alpina used in the present invention (preservation number: CCTCC M2015421) and inoculate them on the potato dextrose agar (PDA) medium plate, 25- Cultivate at 27°C for 10 days until the spores mature, then scrape the spores and hyphae on the potato dextrose agar (PDA) medium plate to fill with 30 ml of sterile water, and shake to obtain a spore suspension.
[0044] b) Shake flask seed culture: inoculate the spore suspension of step (1) into the seed bottle with the culture medium, the inoculum size is 20% (volume ratio), place 30°C, and cultivate 80°C on a shaker at 300 rpm. hours, the medium is: carbon source sucrose 50g / l; nitrogen source yeast extract powder 12g / l; pH 8.5.
[0045] c) Expanded cultivation of seeds: the volume of the final fermenter is 50m 3 , followed by s...
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