Aspergillus niger YH-6 and application thereof in increasing content of icaritin in Herba Epimedii

A technology of YH-6 and Aspergillus niger, applied in the direction of microorganism-based methods, enzymes, biochemical equipment and methods, etc., can solve the lack of economical and effective icariin production methods, long enzymatic hydrolysis reaction time, and low conversion rate and other problems, to achieve the effect of high product yield, low cost, and large increase in content

Active Publication Date: 2020-01-17
安徽箐补堂生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The enzymatic hydrolysis method adopts helicase and naringinase, which has the advantage of high conversion rate, but has the disadvantages of high enzyme cost and long enzymatic hydrolysis reaction time; the acid hydrolysis method has the advantage of low cost, but the hydrolysis process is prone to produce secondary glycosides, resulting in The conversion rate is low, so there is still a lack of cost-effective production methods for icariin

Method used

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  • Aspergillus niger YH-6 and application thereof in increasing content of icaritin in Herba Epimedii
  • Aspergillus niger YH-6 and application thereof in increasing content of icaritin in Herba Epimedii
  • Aspergillus niger YH-6 and application thereof in increasing content of icaritin in Herba Epimedii

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Add about 20 g of Epimedium dried at 85°C and crushed through an 80-mesh sieve into a 250-mL Erlenmeyer flask, then add a small amount of sterile water to moisten it, and incubate at a constant temperature of 28°C for 5 days. Dilute the moldy enrichment 1 x 10 with sterile water 6 After doubling, spread it on the PDA plate medium, culture at 28°C for 3 days, pick out mold colonies with different colors and shapes, transfer them to fresh PDA medium, and place them in 28°C for 3 days to obtain 8 purely cultured strains. They were numbered YH-1 to YH-8 and stored in a 4°C refrigerator for later use.

[0035] The plate medium is potato sucrose agar medium (PDA), prepared according to the following composition and method: wash and peel the potatoes and cut them into small pieces, weigh 200g, add 1000mL of tap water, boil for 30min, filter through 4 layers of gauze to remove slag , the filtrate was made up to 1000mL, then 20g of sucrose and 20g of agar were added, the pH was...

Embodiment 2

[0046] The Aspergillus niger YH-6 bacterial strain obtained by screening in Example 1 is used as the transformation strain, through seed expansion cultivation, fermenting and preparing crude enzyme liquid transformation treatment Epimedium, the specific process steps are as follows:

[0047] (1) Inoculate the Aspergillus niger YH-6 slant strain preserved in a 4°C refrigerator on a fresh PDA plate medium, and culture it in a biochemical incubator at 30°C for 3 days. The composition and preparation method of the plate culture medium are the same as in Example 1.

[0048] (2) Use an inoculation loop to pick up two rings of Aspergillus niger YH-6 spores after activation in step (1) into the seed medium, and culture them at 30°C and 200r / min shaking for 2 days to obtain a seed solution. The composition and preparation method of the seed medium are: sucrose 10g / L, yeast extract powder 5g / L, KH 2 PO 4 2g / L, MgSO 4 0.5g / L, CaCl 2 0.2g / L, the solvent is tap water, pH 6.0; 50mL s...

Embodiment 3

[0053] Using Aspergillus niger YH-6 as the transformation strain, on the basis of Example 2, the composition of the fermentation medium was optimized, the feeding concentration of Epimedium was improved, the transformation time was prolonged, and a preferred transformation process was obtained. The content of icariin can be greatly improved, and the specific process steps are as follows:

[0054] (1) Inoculate the Aspergillus niger YH-6 slant strain preserved in a 4°C refrigerator on a fresh PDA plate medium, and culture it in a biochemical incubator at 30°C for 3 days. The composition and preparation method of the plate culture medium are the same as in Example 1.

[0055] (2) Use an inoculation loop to pick up two rings of Aspergillus niger YH-6 spores after activation in step (1) into the seed medium, and culture them at 30°C and 200r / min shaking for 2 days to obtain a seed solution. The composition and preparation method of the seed medium are: sucrose 10g / L, yeast extrac...

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Abstract

The invention discloses Aspergillus niger YH-6 and application thereof in increasing content of icaritin in Herba Epimedii. The content of icaritin in Herba Epimedii is increased by up to 26.3 times from original 0.273 mg/g to 7.44 mg/g. Compared with an acid hydrolysis method, a method herein has good specificity and high product yield; Compared with pure enzymatic conversion, the method herein has low biocatalyst cost, and allows various glycosides in Herba Epimedii to be converted into icaritin, with the content increased greatly. According to the invention, in-situ conversion of medicinalcomponents in Herba Epimedii is achieved, and the treated Herba Epimedii is used as a raw material for extraction of icaritin, so that the extraction yield is greatly increased.

Description

[0001] (1) Technical field [0002] The invention belongs to the technical field of biochemistry, and in particular relates to a method for increasing the content of icariin in epimedium by adopting a biotransformation method. [0003] (2) Background technology [0004] Icaritin, also known as anhydroicaritin and icaritin, has a CAS number of 118525-40-9 and a molecular formula of C 21 h 20 o 6 , with a molecular weight of 368.4, belonging to flavonoids. Current studies have shown that icariin has estrogen-like properties, immune regulation, anti-inflammation, promotion of cardiomyocyte regeneration and differentiation, promotion of bone protection, promotion of nerve cell differentiation, anti-liver damage and delay of liver fibrosis, anti-tumor, anemia It has the functions of treating and lowering blood sugar, and no obvious toxic and side effects of icariin on normal cells have been found, indicating that it has high safety. As a drug with low toxicity and high efficienc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P17/06C12N9/24C12R1/685
CPCC12N9/2402C12P17/06C12Y302/01C12N1/145C12R2001/685
Inventor 梅建凤王萍萍吴霞应国清易喻
Owner 安徽箐补堂生物科技有限公司
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