Method for carrying out whole-cell catalysis on natural anthocyanin by utilizing microorganisms and transforming natural anthocyanin into protocatechuic acid and application

A technology of protocatechuic acid and microorganisms, which is applied in the field of bioengineering to achieve the effects of simple preparation, improved nutritional benefits, and improved catalytic efficiency

Active Publication Date: 2017-08-18
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method uses anthocyanin as a carbon source, which overcomes the pollution pressure of the existing chemical synthesis and physical extraction technology ...

Method used

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  • Method for carrying out whole-cell catalysis on natural anthocyanin by utilizing microorganisms and transforming natural anthocyanin into protocatechuic acid and application
  • Method for carrying out whole-cell catalysis on natural anthocyanin by utilizing microorganisms and transforming natural anthocyanin into protocatechuic acid and application
  • Method for carrying out whole-cell catalysis on natural anthocyanin by utilizing microorganisms and transforming natural anthocyanin into protocatechuic acid and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-5

[0033] In Examples 1-5:

[0034] The detection conditions of the high-performance liquid chromatography are as follows: 20 μL of sample injection, wavelength of 245 nm, flow rate of 1 mL / min, and mobile phase A: 1% formic acid in water. Mobile Phase B: Acetonitrile. Elution method: 0-4min A phase maintains 95%; 4-16min A phase 95% drops to 88%; 16-30min A phase maintains 70%; 30-35min A phase rises from 70% to 95%.

[0035] The test of the initial concentration of total anthocyanins adopts the pH differential method.

Embodiment 1

[0036] Example 1 Isolation and Identification of Lactobacillus rhamnosus YYJP-2

[0037] 1. Strain Isolation

[0038] (1) Lactobacillus rhamnosus YYJP-2 (L.rhamnosus YYJP-2) was isolated from traditional liquor fermentation koji. The specific separation method is as follows: fully shake the distiller's yeast with sterile distilled water to obtain a suspension, and use the doubling dilution method to dilute the suspension to 10 -3 、10 -4 、10 -5 and 10 -6 The sample solution was spread on the MRS plate and cultivated. After 2 days, the single colony was picked and purified twice by streaking on the plate, and the strains were preserved on the slant for future use.

[0039] (2) The Lactobacillus rhamnosus YYJP-2 (L.rhamnosus YYJP-2) obtained from the above isolation belongs to Gram-positive bacteria, and has large, creamy white, opaque, moist and smooth colonies on the MRS plate. A large amount of viscous carbohydrate material was produced in the broth test tube, manifested ...

Embodiment 2

[0044] The isolation and identification of embodiment 2 Lactobacillus casei KFL2 (L.casei KFL2)

[0045] 1. Strain Isolation

[0046] (1) Lactobacillus casei KFL2 (L.casei KFL2) was isolated from traditional liquor fermentation koji. The specific separation method is as follows: fully shake the distiller's yeast with sterile distilled water to obtain a suspension, and use the doubling dilution method to dilute the suspension to 10 -3 、10 -4 、10 -5 and 10 -6 The sample solution was spread on the MRS plate and cultivated. After 2 days, the single colony was picked and purified twice by streaking on the plate, and the strains were preserved on the slant for future use.

[0047] (2) Lactobacillus casei KFL2 (L.casei KFL2) obtained from the above separation is a Gram-positive bacterium, and has neat, white, opaque, smooth-surfaced colonies on the MRS plate, with a slightly sour taste.

[0048] 2. Strain identification

[0049] The genomic DNA of the above-mentioned isolated b...

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Abstract

The invention discloses a method for carrying out whole-cell catalysis on natural anthocyanin by utilizing microorganisms and transforming the natural anthocyanin into protocatechuic acid. The method is characterized by taking complete cells of specific probiotics as a biological catalyst, taking a weak acidic phosphate buffering solution as a medium and taking a natural anthocyanin extract as a substrate, so as to form a biologically transformed anthocyanin system. The method disclosed by the invention has the advantages that conditions are moderate; the substrate and the product are specific; a subsequent purification procedure is remarkably simplified; the method is a simple and easy strategy for biologically transforming the anthocyanin into the protocatechuic acid by utilizing microorganisms, and other organic matters are not added in a fermentation process, so that possible pollution to environments is reduced. The content of the transformed target product protocatechuic acid can reach 33mg/L at maximum. The method disclosed by the invention has great significance for in vitro study on a mechanism of using microorganism metabolism to transform the anthocyanin, and also provides theoretical basis for directly applying a natural anthocyanin transformed product to probiotic fermented products.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and more specifically relates to a method and application of using microbial whole cells to catalyze the conversion of natural anthocyanins into protocatechuic acid. Background technique [0002] Protocatechuic acid (PCA), namely 3,4 dihydroxybenzoic acid, is an important active ingredient in traditional Chinese medicine. It has been reported abroad that natural anthocyanins are converted into small molecular metabolites - protocatechuic acid in the pig intestine. Protocatechuic acid has many pharmacological effects, mainly antibacterial, anti-tumor, prevention of asthma, and promotion of cortical neuron proliferation in rats. It is clinically used to treat chronic bronchitis. The current industrial production of protocatechuic acid is mainly obtained from vanillin through alkali fusion and acidification. The acid-base substances in this chemical synthesis process will cause certain envi...

Claims

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Application Information

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IPC IPC(8): C12P7/42C12N1/20A23L33/00C12R1/225C12R1/245
CPCA23V2002/00C12P7/42C12N1/205C12R2001/225C12R2001/245A23V2200/30
Inventor 廖振林李汉荣方祥王丽钟青萍
Owner SOUTH CHINA AGRI UNIV
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