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Lactobacillus plantarum with function of reducing contents of biogenic amines in foods and application of lactobacillus plantarum

A technology of Lactobacillus plantarum and biogenic amines, which is applied in the field of microorganisms, can solve the problems of high cost, few types of biogenic amines, and doubtful safety, and achieve good fermentation potential, wide application prospects, and rapid acid production.

Active Publication Date: 2015-12-09
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are still many limitations in the currently isolated microorganisms capable of degrading biogenic amines: (1) There are few types of biogenic amines; (2) The degradation effect is poor; (3) The safety is questionable, etc.
The microorganisms that have been reported to degrade biogenic amines include Halomonas muralis and Saccharomyces cerevisiae engineering bacteria, etc., but their safety in food needs to be further verified, and they do not degrade well for the eight common biogenic amines. role; and the method of adding monoamine oxidase to degrade biogenic amines is not only costly, but also may have higher requirements for equipment

Method used

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  • Lactobacillus plantarum with function of reducing contents of biogenic amines in foods and application of lactobacillus plantarum
  • Lactobacillus plantarum with function of reducing contents of biogenic amines in foods and application of lactobacillus plantarum
  • Lactobacillus plantarum with function of reducing contents of biogenic amines in foods and application of lactobacillus plantarum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 Lactic acid bacteria screening and preservation

[0028] The pickle water of high-quality pickles from the Neijiang area of ​​Sichuan is gradiently diluted with sterile saline, and an appropriate gradient is selected in the MRS broth added with 1.5% bromocresol purple solution (the concentration of bromocresol purple solution is 0.5%) Cultivate on medium, cultivate at 37°C for 48 hours, pick colonies with large color change circles and inoculate them into MRS liquid medium for static cultivation for 24 hours at 37°C. Take 1 mL of the cultured bacterial solution into a bacterial preservation tube, centrifuge at 12,000 rpm at 4°C for 2 minutes, take the supernatant, add 1 mL of 60% glycerol to the precipitate to resuspend, and store at -80°C.

Embodiment 2

[0029] Example 2 Degradation test of Lactobacillus plantarum CGMCCNO.9740 on biogenic amine

[0030] 1. Activation of Lactobacillus plantarum CGMCCNO.9740 strain

[0031] Take the Lactobacillus plantarum CGMCC NO.9740 stored at -20°C and streak it on the MRS plate. After culturing at 37°C for 48 hours, pick lactic acid bacteria cells from well-grown Lactobacillus plantarum colonies and inoculate them into MRS liquid medium for static culture at 37°C for 24 hours.

[0032] 2. Preparation of mother liquor of biogenic amine standard

[0033]The biogenic amines are standard products of biogenic amines, which are purchased from Sigma Company and prepared into a mother liquor with a concentration of 100 mg / L of each biogenic amine during use. The preparation method is: accurately weigh 0.0100mg of tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine, dissolve in 100mL sterile normal saline, mix well and dissolve , and stored at 4°C af...

Embodiment 3

[0053] Embodiment 3 Morphology and identification result of Lactobacillus plantarum CGMCCNO.9740

[0054] Take the Lactobacillus plantarum CGMCC NO.9740 preserved in the glycerol tube at -80°C, and streak it on the MRS plate. After culturing at 37°C for 48 hours, it can be observed that the colony is milky white in shape, smooth and wet on the surface, with smooth edges, see figure 2 , observed under a 40×100 times microscope after Gram staining, it can be seen that Lactobacillus plantarum CGMCC NO.9740 is a rod-shaped Gram-positive bacterium, see image 3 .

[0055] Carry out colony PCR, use a sterile toothpick to pick a single colony into the mixed system, and react in a PCR machine. The system includes: 5 μL of 10×TaqBuffer, 4 μL of 2.5 mMdNTPs, 0.5 μL of 5u / μL Taq DNA polymerase, 0.5 μL of 20 μM upstream and downstream primers (16S universal primers 27F and 1492R), 38.5 μL of sterile water. The reaction program is: 94°C, 5min→(94°C, 30s→50°C, 30s→72°C, 1min)×30 cycles→...

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Abstract

The invention belongs to the technical field of microorganisms and discloses lactobacillus plantarum with a function of reducing contents of biogenic amines in foods and application of the lactobacillus plantarum. The lactobacillus plantarum is resistant to acids and capable of strongly removing eight types of biogenic amines (including tryptamine, phenethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine) in vitro. After 1*1010CFU / ml of the lactobacillus plantarum and the eight types of biogenic amines are co-cultured for 24h, wherein the concentration of each type of the biogenic amines is 100mg / L, and the total amine concentration is 800mg / L, the total amine content is reduced by 70% approximately, the content of each type of the biogenic amines is reduced by 30%-100%, and the removal rate of histamine highest in toxicity is up to 96.69%. Further, the lactobacillus plantarum is capable of lowering pH to 4 in eight hours owing to quickness in acid generation, thereby having favorable fermentation potential. The lactobacillus plantarum is used for degradation of the biogenic amines in foods, especially in fermented foods and extensive in application prospect.

Description

technical field [0001] The invention relates to a plant lactobacillus capable of reducing the content of biogenic amine in food and an application thereof, belonging to the technical field of microorganisms. Background technique [0002] Biogenic amines are a class of biologically active small molecular weight nitrogen-containing compounds mainly formed by the decarboxylation of amino acids or the amination of aldehydes and ketones. They are normal physiologically active substances in organisms and play an important role in body cell activities. However, when the human body consumes excessive biogenic amines from exogenous sources, it will cause headaches, increased blood pressure, damage to the human nervous system and cardiovascular system, and sometimes mainly gastrointestinal dysfunction, causing abdominal cramps, diarrhea and vomiting. Moreover, biogenic amines can react with nitrite to generate carcinogen nitrous compounds. [0003] Biogenic amines are harmful substan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L1/015A23L1/30C12R1/25
Inventor 陈卫田丰伟佟婷婷王刚赵建新张秋香刘小鸣翟齐啸张灏
Owner JIANGNAN UNIV
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