Method for preparing theaflavin by dynamic continuous oxidation on immobilized polyphenol oxidase

A polyphenol oxidase, theaflavin technology, applied in the direction of being fixed on/in an organic carrier, fermentation, etc., can solve the problems of toxicity, waste of enzyme source, difficult separation of product and enzyme, etc., and achieve the content of theaflavin. Stable, easy to control reaction conditions, and the effect of improving economic efficiency

Inactive Publication Date: 2011-10-26
TEA RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chemical oxidation of catechins to prepare theaflavins has the problem of oxidant toxicity, free enzymatic oxidation of tea polyphenols to prepare theaflavins has waste of enzyme sources, separation of products and enzymes is difficult, enzyme reuse cannot be realized, and continuous preparation cannot be achieved, etc. question

Method used

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  • Method for preparing theaflavin by dynamic continuous oxidation on immobilized polyphenol oxidase
  • Method for preparing theaflavin by dynamic continuous oxidation on immobilized polyphenol oxidase
  • Method for preparing theaflavin by dynamic continuous oxidation on immobilized polyphenol oxidase

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Weigh an appropriate amount of immobilized polyphenol oxidase and use the pH 3.2 disodium hydrogen phosphate-citric acid wet packing column prepared with the corresponding tea polyphenol substrate, fill the bottom of the column with a little absorbent cotton, and control the column bed at 100mL. Use disodium hydrogen phosphate-citric acid to prepare 10 mg / mL of tea polyphenol base material solution with pH 3.2 and 40% total amount of catechins, control the temperature of circulating water at 30°C, oxygen flow rate of 55 mL / min, and base flow rate of 2.5 mL / min. Oxygen and the substrate enter the packed bed reactor at the same time to react, and the theaflavin mixture is pumped out continuously with a pump, and the mixture is analyzed in liquid phase. Under this reaction condition, TF is the main body. See figure 1 , and theaflavin content is 16.25%. A theaflavin product with a content of 55% can be obtained by freeze-drying the theaflavin mixture, the freeze-drying pr...

Embodiment 2

[0026] Mix 10mg / mL of tea polyphenols (pH5.7) substrate material solution with 70% total catechins in water, control the temperature of circulating water at 28°C, oxygen flow rate of 70mL / min, and substrate flow rate of 2.0mL / min. Oxygen and the substrate enter the packed bed reactor at the same time to react, and the theaflavin mixture is pumped out continuously with a pump, and the mixture is analyzed in liquid phase. Under this reaction condition, TF is the main body. See figure 2 , and the theaflavin content is 18.39%. A theaflavin product with a content of 70% can be obtained by freeze-drying the theaflavin mixture, the freeze-drying pre-freezing temperature is minus 40° C., and the thickness of the material is 1.5 cm.

Embodiment 3

[0028] Use 30% ethanol to prepare 10 mg / mL of tea polyphenols (pH5.0) base material solution with 80% total catechins, the temperature of circulating water is controlled at 32 °C, the oxygen flow rate is 45 mL / min, and the base flow rate is 3.0 mL / min . Oxygen and the substrate enter the packed bed reactor at the same time to react, and the theaflavin mixture is pumped out continuously with a pump, and the mixture is analyzed in liquid phase. Under this reaction condition, all four main theaflavins appear, see image 3 , and the theaflavin content is 24.57%. The theaflavins mixture was separated on a separation column, and different theaflavins collected fractions were concentrated by rotary evaporation to remove the organic eluent, and the temperature was controlled at 45°C. Then freeze-dry to obtain theaflavin products with different theaflavin contents and ester type ratios, the freeze-drying pre-freezing temperature is minus 50°C, and the material thickness is 1.5cm.

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Abstract

The invention discloses a method for preparing theaflavin through carrying out dynamic continuous oxidation on immobilized polyphenol oxidase. in the method, tea polyphenols is taken as a raw material. At present, the content of theaflavin only accounts for 0.3-1.5% of dry matter of black tea, and the yield is low because the black tea is directly used for extracting. The synthesis and preparation technology of the existing theaflavin can not be dynamically and continuously prepared and is only confined in the laboratory to be prepared, thereby causing higher theaflavin cost and confining the application of theaflavin in medical heath care. In the method, tea polyphenol serves as a substrate, and immobilized polyphenol oxidase serves as a catalyst medium. Reaction temperature is controlled. The substrate and oxygen continuously enter on a packed bed reactor to oxidize tea polyphenol and polymerize to form theaflavin substance, and the theaflavin substance is continuously pumped out by a constant flow pump. In the method, the immobilized polyphenol oxidase is repeatedly utilized on a bioreactor to continuously and dynamically prepare the theaflavin to obtain the theaflavin mixed liquor. The method provides a basis for industrial preparation of theaflavin.

Description

technical field [0001] The invention relates to a method for preparing theaflavins by using tea polyphenols as raw materials and utilizing immobilized polyphenol oxidase dynamic continuous oxidation on a packed bed reactor. Background technique [0002] Theaflavins are a kind of characteristic pigment substances in black tea, which have a great influence on the color and flavor of black tea soup. The degree of fermentation of black tea results in the content of theaflavins only accounting for 0.3-1.5% of the dry matter of black tea. The yield of extraction is very low. Theaflavins are a class of compounds with polyhydroxyl benzotropolone structures, with the following four theaflavins as the main body: [0003] [0004] Theaflavins have many potential and scientifically proven medical and health functions, such as anti-oxidation, anti-cancer, anti-inflammatory activities, etc. At present, the methods for obtaining theaflavins mainly include direct extraction, chemical o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/16C12N11/02
Inventor 江和源王斌张建勇王伟伟
Owner TEA RES INST CHINESE ACAD OF AGRI SCI
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