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Method for preparing fructus arctii aglycone

A technology of arctigenin and arctiin, which is applied in the field of preparation of arctigenin, can solve the problems of complicated process and high cost in the separation and extraction process, and achieve the effect of being environmentally friendly, easy to operate, and realizing large-scale industrial production

Inactive Publication Date: 2009-03-25
ZHEJIANG UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hu Yingjie from Guangzhou University of Traditional Chinese Medicine and others successfully hydrolyzed arctiin into arctigenin by using helicase, which provided an idea for the enzymatic synthesis of arctigenin. The separation and extraction process of pure enzyme is more complicated and costly

Method used

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  • Method for preparing fructus arctii aglycone
  • Method for preparing fructus arctii aglycone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] See Figure 1 for the process flow chart.

[0031] Incline cultivation:

[0032] Preparation of slant medium: Weigh 200g of potatoes, wash, peel and chop them, add 1000ml of water to boil for half an hour, filter with gauze, add 20g of glucose and 20g of agar, fully dissolve, filter with gauze while hot, and pack into test tubes, 1×10 5 Pa sterilize for 30 minutes, place on a slant to cool down, and obtain a slant medium.

[0033] Aspergillus niger CGMCC No.2594 was inoculated into slant medium and cultured at 30°C for 7 days to obtain slant strains.

[0034] Liquid fermentation:

[0035] Liquid fermentation medium preparation (final concentration composition): rice straw powder 50g / L, wheat bran 15g / L, barley flour 15g / L, (NH 4 ) 2 SO 4 10g / L, KH 2 PO 4 0.5g / L, MgSO 4 ·7H 2 O 0.5g / L, initial pH5.0.

[0036] Preparation of β-glucosidase enzyme liquid: inoculate 1ml (the number of spores is about 125 / ml) spore suspension obtained by washing CGMCC No.2594 slant...

Embodiment 2

[0041] According to the method described in Example 1, CGMCC No.2594 cell slant was obtained, and a liquid fermentation medium was prepared.

[0042] Preparation of β-glucosidase enzyme liquid: inoculate 1ml (the number of spores is about 125 / ml) spore suspension obtained by washing CGMCC No.2594 slant strain with physiological saline into a 500ml Erlenmeyer flask containing 200ml fermentation medium Shake at 30°C (150r / min) for 7 days, filter the obtained fermentation broth, adjust the initial pH of the filtrate to 6.0 with HCl and NaOH, and dilute it to 250ml with sterile water as a β-glucosidase enzyme solution for later use. The enzyme activity of the obtained β-glucosidase enzyme liquid is 68U / ml.

[0043] Preparation of the substrate solution: Weigh the arctiin standard substance and dissolve it in 100ml sterile water to prepare 0.6, 1.2, 1.8, 2.4 and 3.0mmol / L substrate solutions respectively.

[0044] Take 10ml of the enzyme solution and mix them with 10ml of the abov...

Embodiment 3

[0048] According to the method described in Example 1, CGMCC No.2594 cell slant was obtained, and a liquid fermentation medium was prepared.

[0049] Preparation of β-glucosidase enzyme liquid: inoculate 1ml (the number of spores is about 125 / ml) spore suspension obtained by washing CGMCC No.2594 slant strain with physiological saline into a 500ml Erlenmeyer flask containing 200ml fermentation medium Shaking (150r / min) at 30°C for 7 days, the obtained fermentation broth was filtered, and the initial pH values ​​of the filtrate were adjusted to 3, 4, 5, 6, 7, 8, 9, 10 and 11 with HCl and NaOH, respectively. The bacterial water was adjusted to 250ml as β-glucosidase enzyme solution for later use. The enzyme activities of the obtained β-glucosidase enzyme solutions were 38, 49, 56, 68, 65, 61, 56, 45 and 35 U / ml, respectively.

[0050] Preparation of the substrate solution: Weigh the arctiin standard substance and dissolve it in 100ml sterile water to prepare 0.6mmol / L substrate...

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Abstract

The invention provides a method for preparing arctigenin, which includes the steps: arctiin is used as a substrate, Beta-glucosidase enzyme liquid produced by the fermentation of aspergillus niger CGMCC No. 2594 is used as a catalyzer, the subsrate and the catalyzer are transformed in a shaker at the temperature of 20 DEG C to 50 DEG C and at the speed of 100r / min to 200r / min for 40h to 64h; and the transformed solution is separated and purified to obtain the arctigenin. The method has the advantages of: 1. mild reaction conditions and being environmental-friendly; 2. simple preparation process of a biocatalyst, namely the Beta-glucosidase enzyme liquid, needing no complex zymoprotein separation and purification as well as low cost; 3. simple operation and relatively high biological transformation efficiency; and 4. easy amplification of the biological transformation process and easy realization of large scale industrialized production. By utilizing the method for producing, the molar yield of the arctigenin can reach 94.7 percent and the purity of the product can reach 99.7 percent.

Description

(1) Technical field [0001] The invention relates to a method for preparing arctigenin, in particular to a method for preparing arctigenin by using β-glucosidase enzyme liquid produced by fermentation of Aspergillus niger CGMCC No.2594 as a biocatalyst. (2) Background technology [0002] Arctigenin ((-)-Arctigenin), the molecular formula is C 21 h 24 o 6 , molecular weight 372.41. Arctigenin has the functions of treating or preventing chronic renal failure and renal fibrosis, anti-virus, anti-tumor, etc. It exists in Arctium in nature. Burdock fruit is the dry ripe fruit of Arctium Lappa L., a plant of Compositae. The effective components of arctium to exert medicinal effects are lignin compounds—arctiin and arctigenin, among which arctiin must first be metabolized into arctigenin before it can be absorbed by the human body. Therefore, arctigenin is an active ingredient that can be directly absorbed by the human body. [0003] There are two main methods for the preparat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/04C12R1/685
Inventor 欧志敏杨根生应国清
Owner ZHEJIANG UNIV OF TECH
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