Pichia pastoris for producing methanol protein and lipase at same time and application thereof
A technology of methanol protein and Pichia pastoris, applied in the field of bioengineering, can solve the problems of outdated fermentation process, single product, high cost, etc., and achieve the effects of simplifying the production process, improving production efficiency and reducing raw material consumption
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[0025] (1) Cultivation of first-class seeds: YPD medium is used as the first-class seed medium, which is divided into 2000mL triangular flasks, with a liquid volume of 500mL per bottle, wrapped with 8 layers of gauze, sterilized at 121°C for 30 minutes, and cooled for later use. Take 3-5 strains of strain zfwx-208 preserved on the slant of the test tube (also known as the slant of the test tube), scrape it off with an inoculation loop and put it into a triangular flask with 500mL of first-level seed medium, inoculate two bottles on the slant of each test tube, and inoculate Afterwards, shaking and culturing in a shaker at 30°C for 22-26 hours, with a shaker speed of 200-300rpm, to obtain first-grade shake flask seeds;
[0026] (2) Cultivation of secondary seeds: Secondary seed medium adopts inorganic salt medium, sterilized at 121°C for 30 minutes and cooled down, then add 4mL of sterilized by 0.22μm microporous membrane filter to each liter of inorganic salt medium Trace elem...
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