Bacillus subtilis strain capable of reducing biogenic amine content, and applications thereof

A technology of Bacillus subtilis and biogenic amines, which is applied to Bacillus subtilis and its application fields, can solve the problems of affecting flavor and not suitable for the production process of bean paste, and achieves the effect of controlling the content of biogenic amines

Active Publication Date: 2017-06-20
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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  • Bacillus subtilis strain capable of reducing biogenic amine content, and applications thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The screening of embodiment 1 subtilis

[0019] 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 the LBGL plate. Aerobic culture 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.

[0020] 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 for 40s, annealing at 56°C for 35s, extension at 72...

Embodiment 2

[0021] Example 2 Degradation of biogenic amine by Bacillus subtilis CGMCC No.13214

[0022] 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.

[0023] 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 the culture medium of Bacillus subtilis, The degradation rates of biogenic amines were: putrescine 59.30±0.59%, cadaverine 68.57±0.48%, histamine 61.92±1.14%, tyramine 63.95±0.51%.

[0024] Simultaneously, with 3% inoculum size (by volume ra...

Embodiment 3

[0025] Example 3 Degradation of Bacillus subtilis CGMCC No.13214 to biogenic amines in fermented soy sauce

[0026] Preparation of sauce unstrained spirits culture: fermented sauce unstrained spirits in the factory for one month; 10mL 1g / L biogenic amine mixture (putrescine, cadaverine, histamine, tyramine), 30g in a 250mL Erlenmeyer flask, sterilized at 121°C for 20min.

[0027] Get 3mL of the bacterium liquid activated by the above-mentioned method and inoculate it into the fermented unstrained spirits culture, and after 7 days of static culture at 37°C, use the Bacillus subtilis CGMCC NO. Cultivate and ferment in the same manner as above. The concentration of remaining biogenic amines and the main flavor substances in the culture were detected by HPLC. Compared with the blank control, the degradation rates of biogenic amines in the sauce fermented grains inoculated with Bacillus subtilis were: putrescine 33.68±0.35%, cadaverine 47.38±0.28%, histamine 15.57±0.22%, tyramine ...

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Abstract

The invention discloses a bacillus subtilis strain capable of reducing biogenic amine content, and applications thereof, and belongs to the technical field of fermentation. The bacillus subtilis strain capable of reducing biogenic amine content is preserved at China General Microbiological Culture Collection Center on 21th, October, 2016, the preservation number is CGMCC No.13214, the preservation address is No.3 zone, No.1 yard, Beichenxi Lu, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences. The bacillus subtilis strain is capable of degrading four kinds of biogenic amines including putrescine, cadaverine, histamine, and tyramine effectively, and the degradation efficiency is as high as 59.30 to 68.57%.

Description

technical field [0001] The invention relates to a strain of Bacillus subtilis capable of reducing biogenic amines and an application thereof, which belongs 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L5/20A23L11/00C12R1/125A23L11/50
CPCC12N1/20C12R2001/125C12N1/205
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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