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Bacillus amyloliquefaciens and application thereof

A technology for dissolving starch spores and bacilli, which is applied in the preparation of bacteria, microorganisms and alcoholic beverages to achieve the effect of enhancing typical flavor characteristics and increasing content

Active Publication Date: 2012-11-07
四川绵竹剑南春酒厂有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some studies have shown that Bacillus amyloliquefaciens can also produce antibacterial substances such as antibacterial protein, iturin (iturin), surfactin (surfactin), fengycin (fetinlin) and bacillomycin (bacillomycin), which have a wide antibacterial spectrum. In particular, it has a good inhibitory effect on mold [4-8] , but there is no report on Bacillus amyloliquefaciens producing pyrazine compounds

Method used

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  • Bacillus amyloliquefaciens and application thereof
  • Bacillus amyloliquefaciens and application thereof
  • Bacillus amyloliquefaciens and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 strain screening

[0031] 1. Primary screening of strains

[0032] Material: distiller's yeast used by Sichuan Jiannanchun (Group) Co., Ltd.

[0033] Medium: 20.0g casein, 1.0g ammonium chloride, 1.0g beef extract, 5.0g magnesium chloride, 20.0g agar, distilled water to 1000mL, pH 7.0

[0034] experimental method:

[0035] Under sterile conditions, take 10 grams of distiller's yeast in a 25 mL Erlenmeyer flask filled with 90 mL of sterile water and glass beads, place it on a bed and vibrate at room temperature for 30 minutes. The bacterial solution was diluted to 10 times in a 10-fold gradient -6 , take 1mL of the bacterial solution of each dilution and put it into a sterile plate, pour it into the sterilized medium cooled to 50°C, mix well, put it into a 37°C incubator after solidification, and cultivate it for 5-7 days, depending on whether it produces The transparent circle and the ratio of the inner and outer diameters of the transparent circle were us...

Embodiment 2

[0046] Identification of embodiment 2TTMP-16 bacterial strain

[0047] 1. 16S rDNA sequence analysis

[0048] (1) DNA extraction

[0049] Collect the bacteria, dissolve in 5mL extraction buffer (100mM Tris Cl, 100mM EDTA-Na 2 , 200mM NaCl, 2%CTAB, pH8.0), shake at 37°C for 45min. Add 0.75mL 20% SDS, and bathe in water at 65°C for 1h. Centrifuge at 12,000rpm for 10min, and collect the supernatant. The supernatant was extracted twice with an equal volume of phenol: chloroform: isoamyl alcohol (25:24:1), added with a final concentration of 0.3M NaAC (pH 5.2) and 2 times the volume of absolute ethanol, and precipitated at room temperature for 1 h. 4°C, 12,000rpm, centrifuge for 20min, collect the precipitate, rinse twice with 70% ethanol, dry and dissolve in 50μL TE (10mmTris-Hcl, 1mmNa 2 EDTA) to obtain total DNA.

[0050] (2) Amplification of 16S rDNA

[0051] Using the total DNA as a template, bacterial 16S rDNA universal primers EU27 (shown in SEQ ID No.1) and 1492R (sh...

Embodiment 3

[0065] Embodiment 3 Morphological characteristics and biological characteristic identification of Bacillus amyloliquefaciens strain of the present invention

[0066] The activated bacteria are used in the following experiments. LB plate medium is used for activation, inoculated by streaking, and cultured upside down at 37°C for 1 to 2 days.

[0067] 1. Observation of colony and bacterial morphology

[0068] Streak inoculate the activated bacteria on LB plate medium, culture at 37°C, observe the colony shape; pick a little bacteria with an inoculation needle, and observe the shape of the bacteria under a microscope (see image 3 ).

[0069] The observation results are as follows: the bacteria are rod-shaped, without capsule, and contain spores; the colonies are cream-colored, opaque, and the edges of the colonies are rough and diffuse.

[0070] 2. Gram staining

[0071] Take a small amount of newly activated bacteria and fix it on a clean glass slide, stain with crystal viol...

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Abstract

The invention relates to a strain of bacillus amyloliquefaciens and an application thereof and belongs to the field of microorganisms. According to the bacillus amyloliquefaciens JNC-002 and the application thereof, the conservation number of the strain is China general microbiological culture collection center (CGMCC) No.5740, and the strain is separated and obtained from distiller's yeast of Sichuan jiannanchun liquor (enterprise group) limited liability company. The capacity of producing tetramethylpyrazine by the bacillus amyloliquefaciens can reach to 2.4g / L, and the bacillus amyloliquefaciens can be applied to producing the tetramethylpyrazine.

Description

technical field [0001] The invention relates to a bacillus amyloliquefaciens and its application, belonging to the field of microorganisms. Background technique [0002] Tetramethylpyrazine is also called Ligustrazine (TTMP for short), and its pharmacological effects mainly include dilating blood vessels; mildly lowering blood pressure; improving tissue microcirculation, increasing tissue blood perfusion; inhibiting platelet adhesion and aggregation and thrombus formation; Proliferation of smooth muscle cells and fibroblasts; regulation of lipid metabolism, anti-lipid peroxidation; certain role in mediating immunity; anti-tissue fibrosis, and certain therapeutic effects on pulmonary fibrosis and liver fibrosis. Pyrazine compounds (3000-6000ug / L in liquor) are unique flavor components of traditional Chinese solid-state fermented liquor, and are also health factors in liquor. Therefore, increasing the content of tetramethylpyrazine in liquor can be beneficial to human health....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P17/12C12G3/02C12R1/07
Inventor 徐姿静唐清兰徐占成樊科权刘孟华焦小川
Owner 四川绵竹剑南春酒厂有限公司
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