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A strain of Bacillus amyloliquefaciens and its application in reducing biogenic amines

A technology of amylolytic spores and biogenic amines, which is applied in the field of fermentation and can solve problems such as affecting the flavor and not being suitable for the production process of bean paste

Active Publication Date: 2019-08-06
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biogenic amine-degrading starter reported so far is mainly isolated from aquatic products and cheese, which is not suitable for the production process of bean paste and may affect the flavor

Method used

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  • A strain of Bacillus amyloliquefaciens and its application in reducing biogenic amines

Examples

Experimental program
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Effect test

Embodiment 1

[0020] The screening of embodiment 1 bacillus amyloliquefaciens

[0021] Take 1g sample from the traditional fermented bean paste fermented bean paste for about 30 days, add it to 9mL sterile water, shake it on a shaker at 30°C at 200rpm for 30min to make a suspension, dilute the suspension gradiently, and spread it on LBGL (LB Culture medium + glucose) plate, cultured aerobically at 37°C for 24h. Pick a single colony without purple halo or a small halo and repeat the plate streak separation, repeat three times to obtain a purified single colony for the next step of identification.

[0022] The -80°C glycerol stock solution was inoculated in LB medium with 1-2% of the inoculum amount, and cultured with shaking at 37°C for 24 hours, and the genomic DNA was extracted by the CTAB method. Use bacterial universal primer pair 27F / 1401R to amplify 16s rDNA. The reaction conditions are: pre-denaturation at 94°C for 3 minutes and then enter the following cycle: denaturation at 94°C fo...

Embodiment 2

[0023] Example 2 Degradation of biogenic amine by Bacillus amyloliquefaciens CGMCC No.13216

[0024] Prepare LBBA medium: peptone 10g / L; yeast powder 5g / L; NaCl 10g / L; putrescine 1g / L; cadaverine 1g / L; histamine 1g / L; tyramine 1g / L, pH natural. Dispense 10mL in a 50mL Erlenmeyer flask, and sterilize at 121°C for 20min.

[0025] Inoculate -80°C glycerol stock solution into fresh LB liquid medium with 1% inoculum amount (by volume ratio), and inoculate on a shaker at 37°C for 12 hours at a rotation speed of 200 rpm. Then inoculate LBBA liquid culture medium with 1% inoculum amount (by volume ratio), cultivate after 4 days, detect the concentration of residual biogenic amine in the culture medium by HPLC method, compare with blank control, inoculate in the culture medium of Bacillus amyloliquefaciens , the degradation rates of biogenic amines were: putrescine 54.32±1.97%, cadaverine 54.58±2.17%, histamine 68.88±1.63%, tyramine 60.28±1.53%.

[0026] With 3% inoculum size (by vol...

Embodiment 3

[0027] Example 3 Degradation of biogenic amines in sauce fermented grains by Bacillus amyloliquefaciens CGMCC No.13216

[0028] Preparation of sauce unstrained spirits culture: Add 10mL of 1g / L biogenic amine mixture (putrescine, cadaverine, histamine, tyramine) to the sauce unstrained spirits that have been fermented in the factory for one month, fill 30g in 250mL Erlenmeyer flasks, and store at 121°C Sterilize for 20 minutes.

[0029] Get 3mL of the bacterium liquid activated by the above-mentioned method and inoculate it in the fermented unstrained spirits culture, 37 ℃ static culture 10 days, adopt bacillus amyloliquefaciens SYB-001 (preservation number is CGMCC No.1314, disclose in CN1683520A) as contrast, according to Cultivate in the same method as above, and detect the remaining biogenic amine concentration in the culture and the physical and chemical indicators and main flavor of the sauce unstrained spirits by HPLC. Compared with the control, the degradation rates o...

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Abstract

The invention discloses a strain of bacillus amyloliquefaciens and its application in reducing biogenic amine, belonging to the technical field of fermentation. The Bacillus amyloliquefaciens of the present invention has been preserved in the General Microorganism Center of the China Microbial Strain Preservation Management Committee on October 31, 2016. The preservation number is CGMCC No.13216, and the preservation address is No. 1, Beichen West Road, Chaoyang District, Beijing. No. 3, Institute of Microbiology, Chinese Academy of Sciences. The strain has a strong performance of degrading biogenic amines, including putrescine, cadaverine, histamine and tyramine, and the degradation efficiency can reach up to 68.88%.

Description

technical field [0001] The invention relates to a strain of bacillus amyloliquefaciens and its application in reducing biogenic amine, belonging to the technical field of fermentation. Background technique [0002] Broad bean paste is fermented and brewed from raw materials such as broad beans, and is a semi-fluid viscous or semi-solid condiment that maintains the original shape of broad bean paste. Broad bean paste has a long history in my country. It is rich in nutrition and delicious in taste, so it is very popular among consumers. However, on the one hand, the current broad bean paste is still prepared by traditional fermentation technology, open fermentation, sun and night dew, and the fermentation strains come from nature, which is uncontrollable and may bring unknown unsafe factors; on the other hand, broad bean Rich in protein, once a large number of free amino acids are metabolized by microorganisms with decarboxylation ability, biogenic amines that are harmful to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L11/00C12R1/07A23L11/50
CPCC12N1/20C12R2001/07C12N1/205
Inventor 李崎刘春凤朱天傲郑飞云李永仙王金晶钮成拓
Owner JIANGNAN UNIV
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