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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

Active Publication Date: 2013-03-13
菏泽建数智能科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, curcumin's poor water solubility and unstable structure limit its application in the fields of food and medicine. Its low selectivity, long-lasting drug effect and poor ability to be absorbed by the body lead to low bioavailability. limit its clinical application
At present, many scholars have synthesized many curcumin derivatives and analogs, and tested and evaluated their physical and chemical properties and biological activities, but there are few reports on the structural modification of curcumin by using microbial cell transformation methods

Method used

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  • Pichia kudriavzevii ZJPH0802 and application thereof in preparation of curcumin derivatives
  • Pichia kudriavzevii ZJPH0802 and application thereof in preparation of curcumin derivatives
  • Pichia kudriavzevii ZJPH0802 and application thereof in preparation of curcumin derivatives

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1: the preparation of curcumin derivative

[0047] (1) Slant culture: Inoculate Pichia kudriazwhizwei ZJPH0802 into the slant medium, and culture at 30°C for 3-5 days to obtain the slant strain; the slant medium is wort medium, and the final concentration consists of Method: Add 20 g of agar to each liter of wort (from Hangzhou Brewery), sterilize at 121°C for 20 minutes at natural pH, cool down after sterilization, and make a slope;

[0048] (2) Seed culture: Pick a ring of bacteria from the slant of step (1) and inoculate it into a 250 mL shake flask containing 100 mL of seed medium, and incubate at 30°C and 200 rpm for 16 hours to prepare a seed liquid; The final concentration of the seed 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, sterilized at 121°C for 20 minutes, cooled after sterilization;

[0049] (3) Fermentation culture: Inoculate th...

Embodiment 2

[0071] Embodiment 2: Orthogonal optimization of fermentation medium components

[0072] Table 2 Orthogonal factor level table of fermentation medium

[0073]

[0074] Utilize Pichia kudriavzevii (Pichia kudriavzevii) ZJPH0802 bacterial strain, after carrying out slant culture and seed culture by the method for embodiment 1, optimize the glucose, yeast extract, NH 4 Cl and MgSO 4 ·7H 2 O concentration. Add the fermented wet cells into a 250 mL Erlenmeyer flask filled with 50 mL of 0.1 M, pH 6.5 phosphate buffer (the amount of wet cells added is 50 g / L based on the dry weight of the cells), and the substrate curcumin concentration 50 mg / L, at 30°C, 200 rpm for 16 h. After the reaction, the cells were removed by centrifugation to obtain the supernatant, which was extracted three times with ethyl acetate, the extracts were combined, and the ethyl acetate was distilled off under reduced pressure at 50°C, and the residue was dissolved in chromatographic methanol to be detecte...

Embodiment 11~16

[0091] Example 11~16 Effect of pH value on the transformation activity of Pichia kudriazwi ZJPH0802

[0092] Utilize the Pichia kudriavzevii ZJPH0802 bacterial strain, after fermenting and culturing according to the method of Example 1, add the wet cells into 50 mL of 0.1M, pH 5.8 to 8.0 phosphate buffer respectively (Table 7) in a 250 mL Erlenmeyer flask (the final concentration of wet cells was 50 g / L), the substrate curcumin concentration was 50 mg / L, and transformed at 30°C and 200 rpm for 16 h. After the reaction, the cells were removed by centrifugation to obtain the supernatant, which was extracted three times with ethyl acetate, the extracts were combined, concentrated under reduced pressure to dryness to remove the ethyl acetate, and the concentrate was dissolved in chromatographic methanol for detection. 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 normalization method, and the results are ...

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Abstract

The invention discloses Pichia kudriavzevii ZJPH0802 and an application thereof in preparation of curcumin derivatives. The strain is collected in China Center for Type Culture Collection (CCTCC), the address is Wuhan University, Wuhan, China, the collection data is September 24, 2012, and the collection number is CCTCC M 2012373. The invention provides a new strain, namely the Pichia kudriavzevii ZJPH0802 for preparing the curcumin derivatives by biotransformation, and structural modification can be performed on curcumin by utilizing a strain resting cell transformation method for obtaining the corresponding derivatives; the pharmacological or biological activity of structurally modified matters of the curcumin can be improved to different extents relative to a curcumin substrate before modification so as to be conductive to development of new pharmaceutical preparations. The technology for obtaining the curcumin derivatives by adopting the biotransformation method, disclosed by the invention, is simpler and environment-friendly, a biological catalyst is a microbial thallus, and the Pichia kudriavzevii ZJPH0802 further has the advantages of self-fermentation and production, stable quality and low cost.

Description

(1) Technical field [0001] The present invention relates to a new curcumin transformation strain----Pichia kudriavzevii ZJPH0802, and its application in the preparation of curcumin derivatives by microbial transformation. (2) Background technology [0002] Curcumin is orange-yellow crystalline powder with slightly bitter taste, insoluble in water, soluble in ethanol, propylene glycol and other organic solvents, easily soluble in glacial acetic acid and alkaline solution, reddish-brown when alkaline, and reddish-brown when neutral and acidic Yellow in color, it is a hydrophobic polyphenolic compound with a special basic skeleton of 1,7-diarylheptane, composed of two ortho-methylated phenols and a β-diketone, and its molecular formula is C 21 h 20 o 6 . Among them, the β-diketone structure has enol-ketone tautomerism, which mainly exists in the structure of ketone formula (1) in acidic and neutral solutions, and mainly exists in the structure of enol formula (2) in alkaline...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12P7/22C12R1/84
Inventor 王普张维宇黄金李军何秀娟
Owner 菏泽建数智能科技有限公司
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