One produces 1α,25(oh) 2 vd 3 media and methods
A culture medium and seed technology, applied in the field of drug biotransformation, can solve the problem of low yield
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Embodiment 1
[0055] Get pseudonocardia (Pseudonocardia autotrophica) CGMCCNo.7935 slant, dig block from the slant and insert in the 250mL shaking flask that 25mL seed culture medium is housed, on shaker 28 ℃, 240rpm cultivates 3 days, obtains seed liquid; Press 10 % Inoculum Amount Inoculate the seed solution into a 250mL shake flask containing 25mL of biotransformation medium, culture it on a shaker at 28°C and 240rpm for 2 days, and take 25mg / mL of 25OHVD 3 Add 1mL of ethanol solution into the biotransformation solution, continue the transformation and culture for 5 days, and then stop the transformation; take 2mL of the transformation solution, add 8mL of methanol, shake for 10 minutes, and centrifuge to get the supernatant for HPLC detection. After determination, 1α,25(OH) in shake flask 2 VD 3 The output reaches 580mg / L.
[0056] The seed medium and biotransformation medium used consist of: peptone 0.5g / l00mL, dextrin 0.5g / l00mL, sodium dihydrogen phosphate 0.2g / l00mL, magnesium sul...
Embodiment 2
[0058] Get pseudonocardia (Pseudonocardia autotrophica) CGMCCNo.7935 slant, dig block from the slant and put in the 250mL shaking flask that 25mL seed culture medium is housed, on shaker 28 ℃, cultivate 3 days at 240rpm, obtain seed liquid, press 10% Inoculation amount Inoculate the seed solution into a 250mL shake flask containing 25mL of biotransformation medium, culture it on a shaker at 28°C and 240rpm for 2 days, and take 25OHVD with a concentration of 25mg / mL 3 Add 1mL of ethanol solution into the biotransformation solution, continue the transformation and culture for 5 days, then stop the transformation; take 2mL of the transformation solution, add 8mL of methanol, shake for 10 minutes, centrifuge to get the supernatant for HPLC detection. After determination, 1α,25(OH) in shake flask 2 VD 3 The output reaches 470mg / L.
[0059] The seed medium and biotransformation medium used consist of: peptone 0.2g / l00mL, dextrin 1.0g / l00mL, sodium dihydrogen phosphate 0.1g / l00mL, ...
Embodiment 3
[0061] Get pseudonocardia (Pseudonocardia autotrophica) CGMCCNo.7935 slant, dig block from the slant and put in the 250mL shaking flask that 25mL seed culture medium is housed, on shaker 28 ℃, cultivate 3 days at 240rpm, obtain seed liquid, press 10% Inoculation amount Inoculate the seed solution into a 250mL shake flask containing 25mL of biotransformation medium, culture it on a shaker at 28°C and 240rpm for 2 days, and take 25OHVD with a concentration of 25mg / mL 3 Add 1mL of ethanol solution into the biotransformation solution, continue the transformation and culture for 5 days, then stop the transformation; take 2mL of the transformation solution, add 8mL of methanol, shake for 10 minutes, centrifuge to get the supernatant for HPLC detection. After determination, 1α,25(OH) in shake flask 2 VD 3 The output reaches 504mg / L.
[0062] The composition of seed medium and biotransformation medium used therein: peptone 1.0g / l00mL, dextrin 0.8g / l00mL, sodium dihydrogen phosphate ...
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