Mortierella alpine mutant strain, method for producing arachidonic acid oil by using same and arachidonic acid oil
A technology of arachidonic acid and Mortierella alpina, which is applied in the field of Mortierella alpina mutant strains, can solve the problems of reducing the content of long-chain saturated fatty acids, not mentioned, etc., 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 CCTCCNO: M2013419 strain as the starting strain, which was deposited by Carbio Bioengineering (Wuhan) Co., Ltd. in China Center for Type Culture Collection (CCTCC) on September 13, 2013. The deposit address is, China , Wuhan, Wuhan University, the deposit number is CCTCCNO: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 move ...
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: CCTCCM2015421) on a potato dextrose agar (PDA) medium plate, and culture at 25-27°C After 10 days until the spores mature, 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; pH7.
[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: Inoculate the commercially available Mortierella alpina and the Mortierella alpina mutant used in the present invention (preservation number: CCTCCM2015421) on a potato dextrose agar (PDA) medium plate, 25-27 Cultivate at ℃ 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 selection volume of 100L,...
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