Pichia kudriavzevii ZJPH0802 and application thereof in preparation of curcumin derivatives
A technology of curcumin derivatives and curcumin, applied in the direction of microorganism-based methods, microorganisms, biochemical equipment and methods, etc., can solve the problems of low bioavailability, poor absorption capacity, and poor water solubility of curcumin
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[0046] Example 1: Preparation of curcumin derivatives
[0047] (1) Slant culture: Inoculate Pichia kudriazwei ZJPH0802 into slant medium, cultivate at 30°C for 3-5 days to obtain slant strain; said slant medium is wort medium with final concentration composition It is: add 20 g agar per liter of wort (from Hangzhou Brewery), natural pH, sterilize at 121°C for 20 minutes, cool after sterilization to make a slope;
[0048] (2) Seed culture: pick a loop of bacteria from step (1) slant bacteria and inoculate it into a 250 mL shake flask containing 100 mL of seed culture medium, cultivate at 30°C and 200 rpm for 16 hours to prepare seed liquid; The final concentration of the seed culture medium is: glucose 25 g / L, yeast extract 2.5 g / L, NH 4 Cl 5.5 g / L, KH 2 PO 4 1 g / L, K 2 HPO 4 1 g / L, MgSO 4 ·7H 2 O 0.4 g / L, solvent is water, pH 6.5, sterilize at 121°C for 20 minutes, then cool after sterilization;
[0049] (3) Fermentation culture: inoculate the seed solution into a 250 mL shake fla...
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[0071] Example 2: Orthogonal optimization of fermentation medium components
[0072] Table 2 Orthogonal factor level table of fermentation medium
[0073]
[0074] Utilize Pichia kudriavzevii ZJPH0802 strain, carry out slant culture and seed culture according to the method of Example 1, and optimize the glucose, yeast extract, and NH in the fermentation medium according to the test plan designed in Table 2. 4 Cl and MgSO 4 ·7H 2 The concentration of O. Add the fermented wet bacteria into a 250 mL Erlenmeyer flask containing 50 mL of 0.1 M, pH 6.5 phosphate buffer (the amount of wet bacteria added is 50 g / L based on the dry weight of the bacteria), and the concentration of the substrate curcumin It is 50 mg / L, converted at 30°C and 200 rpm for 16 h. After the reaction, the bacteria were removed by centrifugation to obtain the supernatant. The supernatant was extracted three times with ethyl acetate. The extracts were combined and distilled under reduced pressure at 50°C to remove e...
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[0091] Example 11~16 The influence of pH value on the transformation activity of Pichia Kudriazwei ZJPH0802
[0092] Using Pichia kudriavzevii ZJPH0802 strain, after fermentation and cultivation according to the method in Example 1, the wet bacteria were added to 50 mL of 0.1M phosphate buffer solution with a pH of 5.8~8.0 (table 7) In a 250 mL Erlenmeyer flask (the final concentration of wet bacteria is 50 g / L), the substrate curcumin concentration is 50 mg / L, and the conversion is carried out at 30°C and 200 rpm for 16 h. After the reaction, the cells were removed by centrifugation and the supernatant was obtained. The supernatant was extracted three times with ethyl acetate. The extracts were combined and concentrated under reduced pressure to dryness to remove ethyl acetate. The concentrate was dissolved in chromatographic methanol for testing. The method is the same as step (5) of Example 1, and the yield of product 6 (t=62 min) is calculated according to the area normalizat...
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