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Aspergillus niger strain for high yield of chlorogenic acid hydrolase and use thereof

A technology of Aspergillus niger strain and chlorogenic acid, which is applied in hydrolytic enzymes, fungi, and microbial-based methods, can solve the problems of unstable yield and conversion rate, lack of basic data in applied research, etc., and achieve easy cultivation and enzyme production High efficiency and fast growth effect

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

AI Technical Summary

Problems solved by technology

[0007] Although the existing technology shows that Aspergillus niger can produce chlorogenic acid hydrolase, the yield and conversion rate reports are very unstable, and there is a lack of basic data on the application of chlorogenic acid conversion and directional hydrolysis to produce caffeic acid and quinic acid

Method used

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  • Aspergillus niger strain for high yield of chlorogenic acid hydrolase and use thereof
  • Aspergillus niger strain for high yield of chlorogenic acid hydrolase and use thereof
  • Aspergillus niger strain for high yield of chlorogenic acid hydrolase and use thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0029] This example illustrates the screening process for Aspergillus niger LN-1.

[0030] Collect soil samples from many plants rich in chlorogenic acid, put them into a conical flask with 50mL of sterile water, add about 20 glass beads, shake vigorously for 20 minutes to completely mash the soil, and let it stand still. Add 1mL of soil sample suspension into each 50mL sterile water Erlenmeyer flask, put it into a constant temperature shaking incubator and cultivate it at 37°C for 2 days. Take the suspension and spread it on the primary screening plate medium to carry out the primary screening of the enzyme-producing strains. The composition of the primary screening medium is: dipotassium hydrogen phosphate 1.0%, potassium dihydrogen phosphate 0.5%, ammonium sulfate 0.1%, sodium nitrate 0.05%, magnesium sulfate 0.05%, ferrous sulfate 0.005%, zinc sulfate 0.005%, agar 5 %, pH6.5, and added 1% chlorogenic acid and 0.002% bromocresol green. The temperature is 35°C, and the cul...

Embodiment 2

[0036] This example illustrates the method of using Aspergillus niger LN-1 to carry out fermentation and enzyme production to produce chlorogenic acid hydrolase, and to obtain caffeic acid and quinic acid by directional enzymatic hydrolysis.

[0037] Medium composition: dipotassium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, ammonium sulfate 0.1%, sodium nitrate 0.01%, magnesium sulfate 0.01%, ferrous sulfate 0.001%, zinc sulfate 0.001%, pH 5.0, and added 0.5 %Chlorogenic acid.

[0038] The spore suspension (concentration) of Aspergillus niger LN-1 was inoculated in the culture medium at an inoculum size of 1%, and cultured aerobically at a temperature of 25° C. for 1 day, that is, the enzyme production ended.

[0039] The chlorogenic acid is dissolved and prepared to be 1%, and the pH is adjusted to 4.0 with NaOH or HCl solution, which is used as the substrate of the enzyme-catalyzed reaction. Add the substrate solution to the crude enzyme solution or wet b...

Embodiment 3

[0041] The method of this embodiment is the same as that of Embodiment 2, and the parameters of the method used are changed.

[0042] Medium composition: dipotassium hydrogen phosphate 1.0%, potassium dihydrogen phosphate 0.5%, ammonium sulfate 0.1%, sodium nitrate 0.05%, magnesium sulfate 0.05%, ferrous sulfate 0.005%, zinc sulfate 0.005%, pH 6.0.

[0043] The spore suspension (concentration) of Aspergillus niger LN-1 was inoculated in the culture medium with an inoculum size of 2%, and cultured aerobically at a temperature of 35° C. for 4 days, that is, the enzyme production ended.

[0044] The chlorogenic acid was dissolved and prepared at 3%, which was used as the substrate of the enzyme-catalyzed reaction. Add the substrate solution to the crude enzyme solution or wet bacteria obtained by filtering the fermentation broth, and adjust the pH to 6.5 with NaOH or HCl solution, at 35°C, with a rotation speed of 150r / min, and react for 24 hours to hydrolyze chlorogenic acid to ...

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Abstract

The invention relates to an Aspergillus niger with a high yield of chlorgenic acid hydrolase screened from plant soil rich of chlorgenic acid, which is nominated as Aspergillus niger LN-1 by classification. The collection unit of the strain is the Center of General Microorganisms, China Committee for Culture Collection of Microorganisms, the collection date is July 14, 2008, and the collection registration number is CGMCC No.2589. The invention further mainly relates to a screening method of the strain, an enzyme production method and the application thereof for the preparation of caffeic acid and quinic acid. The Aspergillus niger LN-1 screened by the method is an aerobic fungus that is easy to be cultivated, produces a plurality of complex enzymes and has the advantages of rapid growth speed, strong reproduction, high-efficiency enzyme production, safeness, high efficiency, the production of chlorgenic acid hydrolase that has activity of orientatedly catalyzing and hydrolyzing the chlorgenic acid so as to compose the caffeic acid and the quinic acid, and high transformation efficiency.

Description

technical field [0001] The invention belongs to the technical field of industrial microbes, in particular to an Aspergillus niger strain with high production of chlorogenic acid hydrolase and its application in preparing caffeic acid and quinic acid. Background technique [0002] Caffeoylquinic acids (Caffeoylquinic acids, CQAs) are the main physiologically active components in plants, and are a phenylpropanoid compound produced by plants through the shikimic acid pathway during aerobic respiration. (Caffeic acid, CA) and quinic acid (Quinic acid, QA) generated depsipolic acid. The most common caffeoylquinic acid is chlorogenic acid (Chlorogenic acid, 5-CQA), also known as coffee tannins (Coffee Tannins), and its hydrolyzate is composed of equimolar caffeic acid and quinic acid. Caffeic acid is a drug approved by my country's SFDA, which has pharmacological activities such as whitening, antibacterial, antiviral, detoxifying, and central nervous system stimulation. Quinic a...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12Q1/04C12N9/18C12P7/42C12R1/685
Inventor 卢定强王俊凌岫泉蒋奔赵辉欧阳平凯
Owner NANJING UNIV OF TECH
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