Method for preparing genistein
A genistein and strain technology, applied in the field of microorganisms, can solve the problems of expensive equipment, high production cost and high technical requirements
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Embodiment 1
[0048] Embodiment 1: Streptomyces ruberus CGMCC No.21851 liquid fermentation culture
[0049] (1) Preparation of seed medium: Thaw and inoculate the suspension of Streptomyces rubrum CGMCC No.21851 frozen in a refrigerator at minus 80°C and inoculate it into the seed medium (the composition of the medium composition: glucose 4 g / L, yeast extract powder 4 g / L, malt extract powder 5 g / L, vitamin B1 g / L, sodium chloride 1 g / L, pH 7.2). Incubate at 29°C for 1 day for inoculation into the fermentation medium.
[0050] (2) Liquid fermentation culture: Add the seed medium to the fermentation medium at a ratio of 10% (soybean meal 30 g / L, glucose 4 g / L, yeast extract powder 4 g / L, vitamin B1 g / L, chlorine NaCl 1 g / L, pH 7.2). After culturing at 29°C for 3 days, a fermentation culture was obtained.
Embodiment 2
[0051] Example 2: Extraction of Streptomyces rubrum CGMCC No.21851 Fermentation and Obtaining of Extract
[0052] The fermented soybean meal fermented by Streptomyces rubrum CGMCC No.21851 in Example 1 was ultrasonically leached three times with the organic solvent ethyl acetate, the extracts were combined, and the extracts were suspended and steamed under reduced pressure to obtain ethyl acetate extract.
Embodiment 3
[0053] Embodiment 3: Separation, preparation and structural identification of genistein
[0054] Get the ethyl acetate extract in embodiment 2 and adopt normal phase chromatographic column to remove main impurity, use 10% dichloromethane-methanol, 50% dichloromethane-methanol and 90% dichloromethane-methanol to elute 2- 3 column volumes, collect the eluate, analyze by TLC or HPLC (eluted with 35% acetonitrile-water solution for 50min), combine the collected solution containing genistein respectively; after removing the eluting solvent by suspension evaporation under reduced pressure, get The crude product containing genistein was purified by semi-preparative HPLC to obtain genistein. The mobile phase used in normal phase HPLC was 35% acetonitrile-water, and the elution peaks containing genistein were collected and subtracted. After suspension steaming or freeze-drying, pure genistein can be obtained.
[0055] Structural identification of the compound of formula I: the compoun...
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