Spironolactone derivative and microbial transformation preparation method and application thereof
A technology of microbial transformation and spironolactone, applied in biochemical equipment and methods, microorganism-based methods, microorganisms, etc., to achieve the effects of easy large-scale cultivation, simple cultivation conditions, and good diuretic function
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Embodiment 1
[0032] Embodiment 1: the preparation of biocatalyst
[0033](1) Strain activation culture: Cunninghamella elegans ATCC9245 (purchased from the American Type Culture Collection) was inoculated into potato agar medium and cultured at a constant temperature of 28°C for 3 days to obtain spores of the fungus. The potato agar (PDA) medium (purchased from EMD Millipore Chemicals, USA) was dissolved in distilled water according to the instructions, and sterilized by high pressure steam at 121°C for 15 minutes;
[0034] (2) Expanded cultivation of seeds: Dip the spores from the activated plate in step (1) with a cotton swab and transfer them to a triangular flask containing 50mL of seed medium, and culture at 28°C and 250r / min constant temperature for 3 day, to obtain the seed solution. The seed medium is potato liquid (PDB) medium (purchased from EMD Millipore Chemicals in the United States), dissolved in distilled water according to the instructions, 50 mL is divided into 250 mL Erl...
Embodiment 2
[0036] The follow-up analysis of the biotransformation product of embodiment 2 spironolactone
[0037] Get 50mL of the cellulite fermented broth of C. silvery mildew ATCC9245 prepared by the method in Example 1, containing 1.64g of wet cellulite; 50mL), transform at 28°C and 250r / min at constant temperature for 4 days. After the transformation culture is over, filter the bacteria with gauze to separate the culture medium. Ultrasonic leaching for 30 minutes, filtering to remove the bacteria, and the filtrate was evaporated to dryness under reduced pressure to obtain a concentrate. The concentrate was dissolved in 0.5mL of anhydrous methanol and filtered to obtain the extract a, which was to be analyzed; the culture solution was extracted with 50mL of ethyl acetate , Ethyl acetate was evaporated to dryness under reduced pressure to obtain a concentrate, which was dissolved in 0.5 mL of anhydrous methanol and filtered to obtain extract b for analysis.
[0038] Under the same con...
Embodiment 3
[0042] Embodiment 3: the preparation of biocatalyst
[0043] (1) Strain activation culture: inoculate Cunninghamella elegans (Cunninghamella elegans) ATCC9245 into potato agar (PDA) medium, and culture at a constant temperature of 28°C for 3 days to obtain the spores of the bacteria. The PDA medium was dissolved in distilled water according to the instructions, and sterilized by high-pressure steam at 121°C for 15 minutes;
[0044] (2) Expanded cultivation of seeds: Dip the spores from the activated plate in step (1) with a cotton swab and transfer them to 4 triangular flasks containing 50mL seed medium, and culture at 28°C and 250r / min constant temperature for 3 day, to obtain the seed solution. The seed medium is PDB medium, dissolved in distilled water according to the instructions, 50mL is divided into 250mL Erlenmeyer flasks, and sterilized by high-pressure steam at 121°C for 15min;
[0045] (3) Bacterial fermentation culture: Inoculate the seed solution prepared in ste...
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