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Method for converting polydatin to resveratrol by microbial enzyme method

A technology of resveratrol glycosides and resveratrol, applied in the field of extraction and transformation of natural active substances

Inactive Publication Date: 2012-04-11
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fundamentally solve the problems of low enzyme catalytic activity, low conversion rate, long time and high cost, and promote the practical application of production

Method used

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  • Method for converting polydatin to resveratrol by microbial enzyme method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1, the acquisition of microbial strains that efficiently transform polydatin into resveratrol:

[0026]Collect wild Polygonum cuspidatum samples from various places, use Polygonum cuspidatum extract to prepare different culture media, screen endophytic bacteria from Polygonum cuspidatum, rinse freshly collected rhizomes of Polygonum cuspidatum under tap water, drain the water, and cut into 2~ For the 3 cm section, perform the following surface disinfection treatment according to conventional aseptic procedures: rinse with 75% alcohol for 3-5 s, rinse with 0.1% mercury liter for 4-5 min, and rinse with sterile water for 4-5 times. Peel off the above-mentioned treated rhizomes under aseptic conditions to take the phloem, cut off the periphery of the phloem, and then cut into small pieces of 0.5 cm × 0.5 cm, and place them on PDA plates. Place 4 pieces on each plate. After incubating at 30°C for 3-7 days, fungal hyphae grew from the cut edge of the sample. After pu...

Embodiment 2

[0027] Embodiment 2, Aspergillus aculeatus ( Aspergillus.aculeatus ) Morphological characteristics of CGMCC No.3876:

[0028] strain Aspergillus aculeatus ( Aspergillus.aculeatus ), CGMCC No.3876 was cultured in PDA medium for 3 to 4 days, and its morphological characteristics were observed with an optical microscope. The mycelium of this strain was colorless, light-colored, or colored substances were aggregated on the surface, and there was a septum. Conidiophores grow vertically from mycelial cells with thick walls and enlarged, most of them have no septum, smooth or rough, the upper part is thicker, and the top expands into spherical, oval, semicircular or club-shaped vesicles; from the vesicles The entire surface of the vesicles produces stalks radially or only at the top of the vesicle; the stalks are single-layered or branched into 2 to more stalks at the top. The conidia are born in clusters on the top of the small stalks, arranged radially or clustered into columns...

Embodiment 3

[0029] Embodiment 3, Aspergillus aculeatus ( Aspergillus.aculeatus ) Fermentation preparation of extracellular enzymes of CGMCC No.3876:

[0030] Prepare medium (formula 4% bran, 1.0% peptone, initial pH 5.0), inoculate Aspergillus aculeatus CGMCC No.3876, and ferment aerobically in a 100 L fermenter at 28-30°C for 3.5 days, and ferment one layer The gauze solution was filtered and centrifuged at 5000 rpm in a three-legged centrifuge for 20 minutes. The supernatant was concentrated 5 times with a hollow fiber membrane with a molecular weight cut-off of 5000 Da. The concentrated solution was 12 L of extracellular enzyme crude enzyme solution compound enzyme; The cellulase activity of bacterial strain, xylanase activity, β-glucosidase activity, the cellulase filter paper enzyme activity (Filter Paper Activity, FPA) of this bacterial strain fermented liquid reaches 3.8 IU / mL (wherein, one FPA enzyme activity International unit refers to the amount of enzyme that generates 1.0 μ...

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Abstract

The invention provides a microbial strain aspergillus.aculeatus (CGMCCNo.3876) used for efficiently extracting polydatin and analogues thereof from plant materials such as polygonum cuspidatum, peanut shells, grape skins and the like, and a complex enzyme prepared by fermenting the microbial strain. The complex enzyme of the strain is used for extracting polydatin and analogues thereof by an enzyme method, and the extraction ratio is 2.3%. The extraction ratio of the complex enzyme is improved by 91.7% through treatment compared with the exoenzyme (control) to which the strain aspergillus.aculeatus is not added. The ratio of conversion from polydatin to resveratrol by using the complex enzyme of the strain is above 99.5%. The polygonum cuspidatum glycoside hydrolase purified from the complex enzyme of the strain is used for converting 98% of polydatin to resveratrol, and the conversion ratio is 99.7%. The purity of resveratrol is 98.5%. By the invention, a novel complex enzyme preparation is actually provided for production, the problems of low catalytic activity of enzymes, low conversion ratio, overlong time and high cost in current production upgrading are fundamentally solved, and the production practice application is promoted.

Description

technical field [0001] The invention relates to a process for transforming Polygonum cuspidatum extract into resveratrol by microbial enzymatic method, and belongs to the technical field of extraction and transformation of natural active substances. technical field [0002] Type the technical field description paragraph here. Background technique [0003] Resveratrol (Res). The scientific name is 3,4,5-trihydroxystilbene, which is an anthraquinone terpenoid compound, and its physical properties are odorless, white crystal powder; it is hardly soluble in water, but easily soluble in organic solvents such as ethanol and acetone. Resveratrol mainly exists in knotweed, grapes, peanuts, pine trees and other plants. Resveratrol has anti-tumor, anti-oxidation, anti-free radical, anti-thrombosis, anti-allergic, anti-atherosclerosis and has the prevention and treatment of coronary heart disease, ischemic heart disease, hyperlipidemia, resveratrol has been Listed as one of the mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/42C12N9/24C12P7/22C12P19/44C12R1/66
Inventor 林元山庞一林邹冬生杨祖佑
Owner HUNAN AGRICULTURAL UNIV
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