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Method for synthesizing theaflavine crude extracts by ultra-fine Cladosporium sp. biology

A technology of biosynthesis and Cladosporium, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of lack of enzyme sources, many reaction by-products, and low enzyme activity in the enzymatic method, and achieve process steps Simple, low equipment investment and low production cost

Inactive Publication Date: 2011-03-09
HUNAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the preparation of theaflavins by in vitro oxidation is mainly through chemical methods or enzymatic methods, but chemical methods have the disadvantages of difficult to grasp the end point and many reaction by-products; enzymatic methods have not been widely used due to problems such as lack of enzyme sources and low enzyme activity. application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1, get soil from tea garden 10cm deep place, take soil sample 5g and put into 95ml sterile water filled with glass beads, shake for 5min, dilute 1000 times, get 0.2ml and inoculate on the primary screening medium plate, Place the plate in a constant temperature incubator at 28°C and incubate for 3 days; streak and inoculate the strains with oxidation circles on the primary screening medium onto the purified medium until the strain of Cladosporium ultrafineii is obtained; then transfer the purified strains to Insert it into the PDA liquid expansion medium, shake it at a speed of 110r / min, and cultivate it at a constant temperature at 23°C for 4 days; 4 NO 3 3g / L, 70.08% purity of large leaf tea catechin 3g / L, CuSO 4 0.1mmol / L, FeSO 4 In the fermented liquid culture medium of 0.1mmol / L, in the container, the filling amount of cultured liquid is 50%, and the inoculum size of bacterial strain is expansion culture medium: the volume ratio of fermentation medium ...

Embodiment 2

[0035]Example 2, take soil from the depth of 15cm in the tea garden, weigh 10g of soil sample and put it into 190ml sterile water filled with glass beads, shake for 5min, dilute 1000 times, get 0.2ml and inoculate it on the medium plate for primary screening , put the plate in a constant temperature incubator at 28°C, and cultivate it for 3 days; streak and inoculate the strains with oxidation circles on the primary screening medium onto the purified medium until the strain of Cladosporium ultrafineii is obtained; and then inoculate the purified strains Transfer to the PDA liquid expansion medium, shake at a speed of 120r / min, and incubate at a constant temperature of 25°C for 4 days; 4 NO 3 3g / L, 76.00% purity of large leaf tea catechin 3g / L, CuSO 4 0.1mmol / L, FeSO 4 In the fermentation medium of 0.1mmol / L, the liquid filling capacity of the culture medium in the container is 60%, and the inoculation amount of the bacterial strain is expansion medium: the volume ratio of fe...

Embodiment 3

[0036] Example 3, take soil from the depth of 20cm in the tea garden, weigh 10g of the soil sample and put it into 190ml sterile water filled with glass beads, shake for 5min, dilute 1000 times, get 0.2ml and inoculate it on the primary screening medium plate, and The plates were placed in a constant temperature incubator at 28°C and cultured for 3 days; the strains with oxidation circles on the primary screening medium were streaked and inoculated onto the purified medium until the strain of Cladosporium ultrafineum was obtained; then the purified strains were transferred to Put it into the PDA liquid expansion medium, shake it at a speed of 130r / min, and cultivate it at a constant temperature of 25°C for 4 days; 4 NO 3 3g / L, 73.87% purity of large leaf tea catechin 3g / L, CuSO 4 0.1mmol / L, FeSO 4 In the fermentation medium of 0.1mmol / L, the liquid filling capacity of the culture medium in the container is 65%, and the inoculum size of the bacterial strain is expansion medi...

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Abstract

The invention relates to a method for biologically synthesizing crude extracts of theaflavins with cladosporium tenuissimum, which comprises following steps: obtaining high-producing strain cladosporium tenuissimum of polyphenol oxidase through screening a large amount of microbes, culturing the strains to synthesize polyphenol oxidase in optimized fermentation medium under the constant temperature and under the condition that the pH value is 4.5-5.6, wherein the rotation speed is 110-130r / min and the temperature is 22-25 DEG C., obtaining crude enzyme liquid of the polyphenol oxidase, and inputting oxygen to simulate oxidation catechin for 40-50 minutes in vitro to prepare the theaflavins. The theaflavins content which is higher than 20.00% in raw products of the theaflavins can be obtained through applying the method of the invention, the process steps are simple, the equipment investment is little, the working efficiency is high, the production cost is low, the method does not use any organic solution in the reaction process, and the method is a green environmental protection theaflavins producing method.

Description

Technical field: [0001] The invention relates to a method for producing theaflavins by microbial extracellular enzymes, in particular to a method for biosynthesizing crude theaflavins by Cladosporium tenuissimum. Background technique: [0002] Theaflavin is the main active substance of black tea, which has a variety of pharmacological and health effects, and is even superior to catechin in some aspects. It has broad application prospects in the fields of natural medicine, functional food, daily chemical industry, and functional beverage development. The currently used theaflavins are mainly separated and extracted from black tea. The total content of theaflavins (TFs) in black tea is low, about 0.2%-2.0%. Technical obstacles have seriously hindered its development and utilization. Obtaining high-content, low-cost theaflavins has become an important research topic in the fields of tea science, food and medicine. In vitro simulated oxidation (including enzymatic oxidation an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12R1/645C12P17/16
Inventor 傅冬和赵淑娟刘仲华
Owner HUNAN AGRI UNIV
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