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A method and application of using microbial whole cells to catalyze the conversion of natural anthocyanins into protocatechuic acid

A technology of protocatechuic acid and anthocyanin, applied in the field of bioengineering, achieves the effects of simple preparation, good enzyme stability, and omission of enzyme purification process

Active Publication Date: 2020-12-08
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 for the preparation of protocatechuic acid, and cannot be directly added to food production. It is also conducive to simplifying the subsequent purification process.

Method used

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  • A method and application of using microbial whole cells to catalyze the conversion of natural anthocyanins into protocatechuic acid
  • A method and application of using microbial whole cells to catalyze the conversion of natural anthocyanins into protocatechuic acid
  • A method and application of using microbial whole cells to catalyze the conversion of natural anthocyanins into protocatechuic acid

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: maintain 95% of phase A in 0-4min; drop 95% of phase A to 88% in 4-16min; maintain 70% of phase A in 16-30min; increase phase A from 70% to 95% in 30-35min.

[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 converting natural anthocyanins into protocatechuic acid by using microbial whole cells to catalyze. The method uses complete cells of specific probiotics as biocatalysts, weakly acidic phosphate buffer as a medium, and natural anthocyanin extracts as a substrate to form a biotransformation anthocyanin system. The method of the invention has mild conditions, specific substrates and products, and significantly simplifies the subsequent purification process. It is a simple strategy for using microorganisms to bioconvert protocatechuic acid to anthocyanins, and no other organic matter is added during the fermentation process, which reduces the impact on the environment. possible contamination. The content of protocatechuic acid converted into the target product can reach up to 33mg / L. The invention is of great significance for in vitro research on the mechanism of microbial metabolic conversion of anthocyanin, and also provides a theoretical basis for the direct application of natural anthocyanin conversion products to probiotic fermented food.

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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Patent Type & Authority Patents(China)
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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