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A kind of method for preparing scutellarein by biotransforming scutellarin

A technology of scutellarin and scutellarein, applied in biochemical equipment and methods, methods based on microorganisms, microorganisms, etc., can solve the problems of low conversion yield, poor specificity, and few by-products, and achieve high conversion efficiency , Strong anti-pollution ability and less by-products

Active Publication Date: 2019-06-14
ZHEJIANG UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for preparing scutellarein by using Aspergillus niger (Aspergillus niger) JH-2 to ferment and prepare crude enzyme liquid to biotransform scutellarin, which can better overcome the existing acid hydrolysis and biotransformation methods for preparing scutellarein The problem of poor performance and low conversion yield, this process has the advantages of low cost, simple process, high conversion yield and less by-products

Method used

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  • A kind of method for preparing scutellarein by biotransforming scutellarin
  • A kind of method for preparing scutellarein by biotransforming scutellarin
  • A kind of method for preparing scutellarein by biotransforming scutellarin

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Experimental program
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Effect test

Embodiment 1

[0019] The selection of embodiment 1 transformation strain

[0020] For the 3 strains of Aspergillus niger in the laboratory, including Aspergillus niger XW-2 (preservation number CCTCC No: M 2011358), Aspergillus niger MJ (preservation number CCTCC NO: M 2013329) and niger Aspergillus JH-2 (Deposit No. CCTCC No: M2014398), and 5 Aspergillus niger strains screened from soil, including Aspergillus niger ZC03, ZC04, ZC08, ZC11 and ZC14 ​​strains. First pick a full cyclospore from the above-mentioned Aspergillus niger slant strain stored in a refrigerator at 4°C, inoculate it in a fresh slant medium, and cultivate the slant in a biochemical incubator at 28°C for 3 days to obtain activated Aspergillus niger slant. Dip the spores from the slant of the activated strains with a cotton swab, inoculate them into 50 mL of enzyme-producing medium, and cultivate them with shaking at 28 °C and 200 r / min for 3 days (the dry cell concentration of the culture solution of different strains is ...

Embodiment 2

[0031] Embodiment 2: verification of transformation stability of Aspergillus niger JH-2

[0032] Using Aspergillus niger JH-2 as the enzyme-producing strain, under the fermentation scale of 50mL shake flask, the step of seed expansion cultivation was added, and the crude enzyme solution was fermented to transform scutellarin, and the transformation stability of the strain was verified. The specific process steps were as follows:

[0033] (1) The aspergillus niger JH-2 slant bacterial classification preserved in 4 ℃ of refrigerators is inoculated in fresh PDA slant medium, and slant is cultivated at 30 ℃ of constant temperature for 2 days, and described PDA slant medium composition and preparation method are the same as embodiment 1 ;

[0034] (2) Use a cotton swab to dip the spores of Aspergillus niger JH-2 after activation in step (1) into 50mL seed culture medium twice, and cultivate them at 30°C and 200r / min constant temperature shaking conditions for 2 days to obtain a dry...

Embodiment 3

[0038] Embodiment 3: preferred conversion process

[0039] On the basis of Example 2, the composition of the enzyme-producing medium, fermentation and culture conditions, conversion substrate concentration, and conversion time were optimized. Aspergillus niger JH-2 was fermented to prepare the crude enzyme solution for the conversion of scutellarin, and the conversion of scutellarein was obtained rate increased significantly. The preferred biotransformation method prepares the processing steps of scutellarein as follows:

[0040] (1) Inoculate the Aspergillus niger JH-2 slant bacterial classification preserved in 4 DEG C of refrigerators on fresh slant medium, and culture the slant at a constant temperature of 30 DEG C for 2 days, and the composition and preparation method of the slant medium are the same as in Example 1;

[0041] (2) Use a cotton swab to dip the spores of Aspergillus niger JH-2 after activation in step (1) into 50mL seed medium for 2 times, and cultivate the...

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Abstract

The invention discloses a method for preparing scutellarein by biotransformation of scutellarin. In the method, the filtrate obtained by suction filtration of the fermented liquid after Aspergillus niger JH-2 enzyme-producing culture is used as a biocatalyst, and scutellarin is used as the biocatalyst. For the substrate, methanol is used as a co-solvent to form a transformation system, and the transformation reaction is carried out under the condition of constant temperature oscillation at 30-35°C. After the transformation reaction is completed, the transformation liquid is separated and purified to obtain scutellarein; the feeding concentration of the substrate is 2g / L , the total conversion yield of scutellarein was 85.6%, and the purity was 82.3%. Aspergillus niger JH‑2 has low nutritional requirements, short culture time, and strong anti-pollution ability; it grows fast and has high conversion efficiency; no harmful substances are produced during the conversion process , few by-products; the invention converts scutellarein with low activity into scutellarein with better activity, which can be applied in large-scale industrialization, and the production process has the advantages of short cycle, high conversion rate, and low environmental pollution.

Description

(1) Technical field [0001] The invention belongs to the technical field of biochemical engineering, and in particular relates to a method for preparing scutellarein by biotransforming scutellarin. (1) Background technology [0002] Scutellarein (scutellarein), also known as high baicalein, baicalein and baicalein, etc., has a CAS number of 529-53-3 and a chemical structure of 4',5,6,7-tetrahydroxyflavone (4', 5,6,7-tetrahydroxy-flavone), the molecular formula is C 15 h 10 o 6 , with a molecular weight of 286.24, belongs to flavonoids and has no optical activity. The pure product is light yellow needle-like crystal, the molecule is planar, insoluble in water, soluble in hot methanol, methanol-chloroform, dimethyl sulfoxide and other solvents, because it contains 4 phenolic hydroxyl groups, it is weakly acidic. The 7-O-β-glucuronide of scutellarein, namely breviscapine (breviscapine), also known as scutellarin, scutellarin and baicalin, etc., the CAS number is 27740-01-8, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/06C12R1/685
CPCC12P17/06C12N1/145C12R2001/685
Inventor 郑晨沈琪华梅建凤董志红王晔易喻应国清
Owner ZHEJIANG UNIV OF TECH
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