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Strain S2-16 for growing and secreting secondary metabolites of chlorogenic acid and application thereof

A technology of secondary metabolites and chlorogenic acid, applied in the direction of microorganism-based methods, chemicals for biological control, applications, etc., can solve the problems of many limiting factors and unstable yield of chlorogenic acid, and achieve the goal of suppressing Bacteria effect is remarkable, the effect of increasing yield

Active Publication Date: 2018-06-22
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the production of chlorogenic acid is subject to many restrictive factors, and the output is unstable.

Method used

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  • Strain S2-16 for growing and secreting secondary metabolites of chlorogenic acid and application thereof
  • Strain S2-16 for growing and secreting secondary metabolites of chlorogenic acid and application thereof
  • Strain S2-16 for growing and secreting secondary metabolites of chlorogenic acid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 : Isolation and Identification of Colletotrichum acutatum S2-16 (Colletotrichum acutatum)

[0036] 1. Isolation of Anthrax oxysporum S2-16 (Colletotrichum acutatum)

[0037] The separation of Anthrax oxysporum S2-16 (Colletotrichum acutatum) includes three steps of sampling, primary screening and secondary screening, as follows:

[0038] 1.1 Sampling

[0039] To isolate chlorogenic acid-producing strains, fresh plant leaves were collected at Guangxi University (N22°50'28.41", E108°17'9.00") and Guangxi Medicinal Botanical Garden (E108°19', N22°51'). The samples were put into sample bags one by one and stored in a refrigerator at 4°C.

[0040] 1.2 Primary screening

[0041]Using the tissue block method, take the plant leaves and wash them with sterile water, wipe them clean with sterile filter paper, place them in an ultra-clean workbench, sterilize them in 75% alcohol for 60-90 seconds for several times, rinse them with sterile water several times, and th...

Embodiment 2

[0055] Example 2 : Qualitative Analysis of Chlorogenic Acid Production by Anthrax oxysporum S2-16 (Colletotrichum acutatum)

[0056] 1.1 Qualitative leaching and thin layer chromatography (TLC) of Anthrax oxysporum S2-16 (Colletotrichum acutatum)

[0057] The strains were selected for shaking-table fermentation and cultured for about 5 days, and the pH of the fermentation broth was adjusted to 4.0-5.0 to maintain the stability of chlorogenic acid. First, use a cell disruptor to break the cells and ultrasonically disrupt for 30 minutes to release the cell contents; then, centrifuge to take 1 mL of the supernatant, add 75% ethanol at a ratio of 1:1 and let it stand for 5 hours, then use a rotary evaporator to concentrate in vacuo at 45°C for about 30 minutes , to speed up the liquid-to-mass conversion to speed up the extraction efficiency; finally, add an equal volume of ethyl acetate to extract 3 times, each time take the ethyl acetate layer and concentrate it in vacuum until...

Embodiment 3

[0066] Example 3 : Quantitative analysis of chlorogenic acid produced by crude extract of Anthracnose oxysporum S2-16 (Colletotrichum acutatum)

[0067] 1.1 Qualitative and quantitative analysis of Anthrax oxysporum S2-16 (Colletotrichum acutatum) by liquid chromatography-mass spectrometry (UPLC-MS)

[0068] The sample was prepared into a sample solution to be tested according to the chlorogenic acid extraction method.

[0069] UPLC was performed using an Agilent Co system equipped with a binary solvent delivery system, autosampler and PDA detector. On an Agilent column (ZORBAX RRHD Eclipse Plus C 18 Chromatographic analysis was carried out on a column; 2.1 mm×50 mm, 1.8 μm; Agilent Corporation, Santa Clara, California, United States). The mobile phase consisted of (A) 0.5% acetic acid in water and (B) acetonitrile. Optimize the UPLC elution conditions as follows: 90% A and 10% B (0 ~ 5min), 75% A and 25% B (5 ~ 6min), and 90% A and 10% B (6 ~ 9min); the flow rate is 0.3 ...

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Abstract

The invention belongs to the technical field of microbial application, particularly relates to a strain S2-16 for growing and secreting secondary metabolites of chlorogenic acid and application thereof. The strain S2-16 for growing and secreting secondary metabolites of chlorogenic acid has the taxonomy name of Colletotrichum acutatum S2-16, and is collected in CCTCC (China Center For Type CultureCollection) on December 14, 2017. The preservation number of CCTCC NO: M 2017789. A coarse extract of the strain S2-16 can inhibit gram-positive bacteria and gram-negative bacteria, and can realize the anti-oxidization and free radical clearing effects; the DPPH clearing rate reaches 77.7 percent.

Description

technical field [0001] The invention belongs to the technical field of microorganism application, and in particular relates to a bacterial strain S2-16 capable of growing and secreting secondary metabolite chlorogenic acid and its application. Background technique [0002] Chlorogenic acid (CGA) is an important secondary metabolite produced through shikimic acid and phenylalanine pathways in the process of plant aerobic respiration, and widely exists in higher dicots and ferns. It is dehydrated and condensed from caffeic acid and quinic acid to 3-O-caffeoylquinic acid (3-O-caffeoylquinic acid), the molecular formula is C 6 h 18 o 9 , belonging to polyphenolic organic acids. The medicinal function of chlorogenic acid is outstanding, mainly related to anti-oxidation and scavenging free radicals, significantly improving the activity of glutathione reductase (GSH-Px) and catalase (CAT); antibacterial, anti-tumor, and significantly inhibiting tumor Cells (MCF-7) proliferated ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12P7/62A01N37/38A01P1/00A01P3/00A01P21/00C12R1/645
CPCA01N37/38C12P7/62C12N1/145C12R2001/645
Inventor 樊宪伟王晓李有志
Owner GUANGXI UNIV
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