Method for producing 17 alpha-methyl-teslosterone by employing spherical arthrobacterium
A technology of Arthrobacter globosa and hydrogen methyl testis, which is applied in the field of microbial pharmacy and medical engineering, can solve the problems of restricting large-scale industrial production, lengthy process steps, difficult quality assurance, etc., and achieves easy separation and purification, and specific reaction Good, the effect of a single product
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Embodiment 1
[0032] A production method of 17α-dehydromethyltestosterone, the steps of the production method are:
[0033] (1). First-level seed culture: Arthrobacter sphaericus (A.Globiformis ATCC 31250) is used as the production strain to connect a full ring from the slope to 30ml seed medium, on a rotary shaker with a speed of 160r / min and a temperature of 30°C Cultivate under the conditions for 22 hours to obtain a seed culture solution suitable for inoculation.
[0034] (2). Secondary fermentation culture: insert the seed culture solution into the fermentation medium with an inoculation amount of 5% volume ratio, control the shaking table speed at 180r / min during the fermentation process, and cultivate at 32°C for 24 hours , to obtain cell fermentation liquid.
[0035] (3). Transformation: Grind 17α-methyltestosterone into fine powder, with a particle size of 14μ, and put it into the bacterial fermentation broth under aseptic conditions, and the feeding amount is 5g / L; then add hydro...
Embodiment 2
[0038] (1). Primary seed culture: Arthrobacter sphaericus (A.Globiformis ATCC 31250) is connected to a 30ml seed medium from a slant, and cultured at 32°C for 18 hours on a rotary shaker at 180r / min. Promptly obtain the seed culture solution suitable for inoculation;
[0039] (2).Secondary fermentation culture: insert the well-grown seed culture solution into 30ml fermentation medium at an inoculum size of 3% (v / v); control the shaking table speed 170r / min during the fermentation process, 30°C Cultivate for 22 hours to obtain the cell fermentation liquid;
[0040] (3). Transformation: Grind 17α-methyltestosterone into fine powder, with a particle size of 15μ, and put it into the fermentation broth of bacteria under aseptic conditions, and the feeding amount is 6g / L; then add hydrophilic to the fermentation broth of bacteria 3% ethanol as a neutral organic solvent; in C 1,2 During the dehydrogenation conversion process, the rotation speed of the shaker was controlled at 190 r...
Embodiment 3
[0043] (1). Primary seed culture: Arthrobacter sphaericus (A.Globiformis ATCC 31250) is connected to a full ring from the slope to 30ml seed medium, and cultivated at 31°C for 20 hours on a rotary shaker at 170r / min. Promptly obtain the seed culture solution suitable for inoculation;
[0044] (2). Secondary fermentation culture: insert the well-grown seed culture solution into 30ml fermentation medium at an inoculum size of 4% (v / v); control the shaking table speed 160r / min during the fermentation process, 33 Cultivate for 18 hours at ℃ to obtain the cell fermentation broth;
[0045] (3). Transformation: Grind 17α-methyltestosterone into fine powder, with a particle size of 16μ, and put it into the bacterial fermentation broth under aseptic conditions, and the feeding amount is 8g / L; then add hydrophilic to the bacterial fermentation broth Proactive organic solvent 5% dimethyl sulfoxide, in C 1,2 During the dehydrogenation conversion process, the rotation speed of the shaker...
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