11-alpha hydroxylation reaction preparation method of steride hormone substance important intermediate
A technology for steroid hormones and intermediates, which is applied in the field of bioengineering transformation, can solve the problems of complex and tedious processes and high production costs, and achieve the effects of shortening reaction time, simplifying preparation method and shortening total reaction time.
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Embodiment 1
[0026] (1) The first step is to cultivate the seeds of Metarhizium anisopliae. Firstly, the slant medium and seed medium are prepared and sterilized. The mold strain is spread on the slant medium and cultivated at 27 degrees Celsius for 5-7 days. Add sterile water to the slant of the bacteria, and sweep it down with a cotton swab to make a concentration of about 1.2×10 8 spores / mL. Take 0.5ml of spore liquid, inoculate it into 100mL of sterilized seed medium, and cultivate it for about 24 hours at 28 degrees Celsius and 150-200r / min. Among them, the composition of the seed medium (g / L): 0.05 dipotassium hydrogen phosphate, 0.05 magnesium sulfate, 0.005 ferric ammonium citrate, 2 citric acid, 2 ammonium nitrate, 10 calcium carbonate, 20 glycerol, and 5 glucose. Slant medium (g / L): Add 15g of agar on the basis of the seed medium.
[0027] (2) The second step is to carry out the cultivation of the shake flask fermentation medium, transplant 80ml of the cultured seed liquid into...
Embodiment 2
[0032] (1) The first step is to cultivate the seeds of Ochraus japonicus. First, prepare the slant medium and seed medium, and sterilize it. Take the mold strain and spread it on the slant medium, and cultivate it at 25 degrees Celsius for 5-7 days. Add sterile water to the slant of the bacteria, and sweep it down with a cotton swab to make a concentration of about 1.2×10 8 spores / mL. Take 0.5ml of spore liquid, inoculate into 100mL sterilized seed medium, and cultivate for about 24h at 27 degrees, 150-200r / min. Among them, the composition of the seed medium (g / L): 0.05 dipotassium hydrogen phosphate, 0.05 magnesium sulfate, 0.005 ferric ammonium citrate, 2 citric acid, 2 ammonium nitrate, 10 calcium carbonate, 20 glycerol, and 5 glucose. Slant medium (g / L): Add 15g of agar on the basis of the seed medium.
[0033] (2) The second step is to carry out the cultivation of the shake flask fermentation medium, transplant 80ml of the cultured seed liquid into the sterilized 1600ml...
Embodiment 3
[0038] (1) The first step is to cultivate the seeds of Trichoderma and Metarhizium anisopliae. First, configure the slant medium and seed medium, and sterilize it. Take the mold strain and spread it on the slant medium, and cultivate it at 32 degrees Celsius for 5 to 7 days. . Add sterile water to the slant of the bacteria, and sweep it down with a cotton swab to make a concentration of about 1.2×10 8 spores / mL. Take 0.5ml of spore liquid, inoculate into 100mL sterilized seed medium, and cultivate for about 24h at 32 degrees, 150-200r / min. Among them, the composition of the seed medium (g / L): 0.05 dipotassium hydrogen phosphate, 0.05 magnesium sulfate, 0.005 ferric ammonium citrate, 2 citric acid, 2 ammonium nitrate, 10 calcium carbonate, 20 glycerol, and 5 glucose. Slant medium (g / L): Add 15g of agar on the basis of the seed medium.
[0039] (2) The second step is to carry out the cultivation of the shake flask fermentation medium, transplant 80ml of the cultured seed liqu...
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