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Aspergillus niger capable of producing glycosidase and application thereof in improving resveratrol content in Japanese knotweed

A technology of Aspergillus niger and glycosidase, applied in the field of glycosidase-producing bacteria, can solve the problems of low conversion rate, complicated operation, easy pollution, etc., and achieve the effects of improved extraction yield, simple nutritional requirements and rapid growth

Active Publication Date: 2012-07-04
ZHEJIANG UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is simple, but conversion rate is low, easily pollutes, and fresh knotweed cuspidatum is inconvenient to preserve in the production process, and mass production is difficult; enzymatic hydrolysis at temperature
This method has good specificity, but the cost of using commercial enzymes is high, and the crude extraction of polydatin is cumbersome and inefficient; (3) Polygonum cuspidatum is pulverized and then added with water, inoculated with certain microorganisms, or natural fermentation without inoculation
This method has the advantages of simple method and low cost, but the microbial cells and their metabolites remain in the Chinese medicinal materials, which affects the quality of the medicinal materials

Method used

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  • Aspergillus niger capable of producing glycosidase and application thereof in improving resveratrol content in Japanese knotweed
  • Aspergillus niger capable of producing glycosidase and application thereof in improving resveratrol content in Japanese knotweed
  • Aspergillus niger capable of producing glycosidase and application thereof in improving resveratrol content in Japanese knotweed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Enrichment, isolation and screening of glycosidase-producing microorganisms

[0029] Add about 50g of Polygonum cuspidatum dry powder (passed through an 80-mesh sieve after crushing) into a 250mL conical flask, add an appropriate amount of tap water and 5g of pastoral soil, stir evenly, and place it in a 30°C incubator for 7 days to enrich for glycosidase-producing microorganisms.

[0030] The above-mentioned enrichment overgrown with mold was diluted with sterile water and spread on a plate medium, and cultured in a biochemical incubator at 30° C. for 48 hours. The mold colonies with different shapes and colors were picked and inoculated on the slant medium, and the slant was placed at a constant temperature of 30°C for 48 hours to obtain spore-rich slant strains.

[0031] Pick the spores of the above strains with an inoculating loop, insert them into 50 mL of enzyme-producing medium (packed in a 250 mL triangular flask), and ferment them for 48 hours at 30°...

Embodiment 2

[0035] Example 2: Fermentation for Bioconversion of Polygonin to Resveratrol Glycosidase

[0036] Taking Aspergillus niger XW-2 as an enzyme-producing strain, after the optimization of the composition of the medium and the fermentation conditions, the activity of the glycosidase in the crude enzyme liquid is greatly improved compared with the activity of Example 1, and the preferred preparation method is as follows:

[0037] (1) Inoculate the Aspergillus niger XW-2 strain stored on the slant of the test tube into the slant medium, and culture the slant in a 30°C biochemical incubator for 48 hours. The slant culture medium is prepared according to the following composition and method: the potatoes are washed and peeled, cut into small pieces, 200 g are weighed, 1000 mL of tap water is added and boiled for 30 min, the potato pieces are filtered off with 4 layers of gauze, the filtrate is made up to 1000 mL, and then glucose is added. 20g, NaCl 5g, agar 20g, natural pH, heat unti...

Embodiment 3

[0042] Example 3: Biotransformation method improves resveratrol content in Polygonum cuspidatum

[0043] The rhizomes of the commercially available Chinese medicinal material Polygonum cuspidatum were dried in an oven at 85°C for 24 hours, pulverized with a pulverizer, and passed through an 80-mesh sieve. Put 50 mL of the crude glycosidase solution prepared in Example 2 in a 250 mL conical flask, add 5 g of Polygonum cuspidatum dry powder, seal the conical flask with plastic wrap, and then biotransform for 24 h in a water bath shaking box at 30°C and 300 r / min. The transformation product in the triangular flask was filtered with a Buchner funnel, and the Polygonum cuspidatum powder was dried in an oven at 85° C. to obtain the biotransformed Polygonum cuspidatum. The content of resveratrol in the biotransformed Polygonum cuspidatum was analyzed by high performance liquid chromatography, reaching 0.92% ( image 3 ), while the content of resveratrol in Polygonum cuspidatum witho...

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Abstract

The invention provides a bacterial strain capable of producing glycosidase, namely Aspergillus niger XW-2, and application thereof in improving resveratrol content in Japanese knotweed in the biotransformation method. The bacterial strain is preserved in the Chinese typical culture preservation center, the preservation number is CCTCC No: M2011358, and the preservation date is 2011 / 10 / 19. The Aspergillus niger capable of producing glycosidase and the application thereof in improving resveratrol content in Japanese knotweed have the advantages that: (1) Aspergillus niger is normally-used strains in the fermentation industry and is non-toxic, harmless and safe to use; (2) Aspergillus niger XW-2 is simple in nutrition requirements, fast in growth and easy to culture; (3) glycosidase producedby Aspergillus niger XW-2 is extracellular enzyme, after thalli are removed through separation, coarse enzyme liquid directly serves as a biotransformation system, and after Japanese knotweed is grinded and added into the coarse enzyme liquid for transformation, content of resveratrol in Japanese knotweed can be improved by at most four times; and (4) Japanese knotweed after biotransformation treatment contains no thalli or metabolite residues of thalli, so that Japanese knotweed can be used as Chinese herbal medicines and can be used for extraction and preparation of resveratrol.

Description

(1) Technical field [0001] The invention relates to a strain of glycosidase-producing bacteria-Aspergullis niger XW-2, and its application in increasing the content of resveratrol in Polygonum cuspidatum through biotransformation. (2) Background technology [0002] Polygonum cuspidatum, a traditional Chinese medicine, is the dried rhizome of Polygonum cuspidatum. The main active ingredients are stilbenes and anthraquinones. The former includes polydatin and resveratrol. Polygonum cuspidatum is also called resveratrol glycosides . Resveratrol is an active polyphenolic substance, which has the functions of inhibiting tumors, anti-oxidation, anti-free radicals, anti-thrombosis, anti-allergy, anti-atherosclerosis and prevention of coronary heart disease, ischemic heart disease, hyperlipidemia, etc. Resveratrol has been listed as one of the most promising anti-cardiovascular and anti-cancer drugs. The content of resveratrol in dried Polygonum cuspidatum rhizome is only 0.1%-0.2...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N9/24C12P7/22C12R1/685
Inventor 梅建凤许南儿罗溜彬王琼芝应国清王鸿
Owner ZHEJIANG UNIV OF TECH
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