Biological transformation method by utilizing hydrophobic compound of cyclodextrin
A biotransformation and cyclodextrin technology, applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., can solve the problems of long biotransformation time, limited bioavailability, low conversion efficiency, etc.
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Embodiment 1
[0076] Compound C of cortisone acetate 1,2 dehydrogenation reaction
[0077] Microbial strain
[0078] The microbial strain adopts Arthrobacter sp.CICC 10193, which can realize the C 1,2 The conversion of cortisone acetate to prednisone acetate by dehydrogenation.
[0079] Incline medium (g / L): glucose 10, yeast extract 10, agar 20, pH7.0
[0080] Seed medium (g / L): glucose 10, corn steep liquor 10, peptone 5, dipotassium hydrogen phosphate 2.5, pH 7.0
[0081] Fermentation medium (g / L): glucose 10, corn steep liquor 15, peptone 5, dipotassium hydrogen phosphate 2.5, pH 7.0
[0082] Cultivation of slant seeds: take a ring of slant strains and transfer to fresh slant medium, and culture at 32°C for 24 hours.
[0083] Bacterial culture:
[0084] Take 5mL of sterile water to wash the slanted seeds to prepare a bacterial suspension. Transplant into 30mL seed medium according to 1% inoculum amount, culture at 32°C and shake at 160r / min for 24h.
[0085] The seed culture sol...
Embodiment 2
[0090] Embodiment 2 is basically the same as Example 1
[0091] Compound C of cortisone acetate 1,2 dehydrogenation reaction
[0092] The conversion substrate cortisone acetate is ground into a fine powder, and is directly thrown into the fermentation medium at a concentration of 4% of the weight of the fermentation broth, then 5% industrial alcohol by volume of the fermentation medium is added, and the substrate acetic acid can be added simultaneously. The sulfobutyl ether-β-cyclodextrin with a pine molar ratio of 0.3:1 was fermented at 34° C. and 160 r / min for 50 h.
[0093] Heat the culture solution to a slight boil, stop the transformation, and let it cool down to room temperature naturally for analysis.
[0094] External standard method detection. The conversion rate was determined to be 98.7%. The conversion rate of the blank control sample without adding was 92.6% in the simultaneous experiment under the same conditions.
Embodiment 3
[0095] Embodiment 3 is basically the same as Example 1
[0096] Compound C of cortisone acetate 1,2 dehydrogenation reaction
[0097] The conversion substrate cortisone acetate is ground into a fine powder, and is directly thrown into the fermentation medium at a concentration of 2.5% of the weight of the fermentation broth, and then 4% industrial alcohol by volume of the fermentation medium is added, and the substrate acetic acid can be added simultaneously. The hydroxypropyl-β-cyclodextrin with a pine molar ratio of 0.25:1 was fermented at 34° C. and 160 r / min for 50 h.
[0098] Heat the culture solution to a slight boil, stop the transformation, and let it cool down to room temperature naturally for analysis.
[0099] External standard method detection. The conversion rate was determined to be 98.9%. The conversion rate of the blank control sample without adding was 93.7% in the simultaneous experiment under the same conditions.
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