Clostridium sporogenes and purpose thereof

A Clostridium sporogenes, the use of technology, applied in the field of Clostridium sporogenes, to achieve the effect of improving the rate of high-quality products, large application and promotion value

Active Publication Date: 2014-01-22
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

[0003] After the 1970s, liquor technicians made a lot of reports on the isolation, cultivation and application of caproic acid bacteria, but most of them focused on productive practical technologies, while the research on microbiology, especially the strains Relatively little research has been done on the identification and physiology of

Method used

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  • Clostridium sporogenes and purpose thereof
  • Clostridium sporogenes and purpose thereof
  • Clostridium sporogenes and purpose thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 strain screening

[0026] 1 strain screening

[0027] 1.1 Primary screening of strains

[0028] 1.1.1 Material: Jiannanchun old cellar mud

[0029] 1.1.2 Medium:

[0030] Sodium acetate 8g, MgC 2 200mg,NH 4 Cl500mg, MnSO 4 2.5mg, CaSO 4 10mg, FeSO 4 5mg, sodium molybdate 2.5mg, biotin 5ug, p-aminobenzoic acid 100ug, distilled water 1000mL, natural pH, sterilize at 121°C for 20min, after cooling, add ethanol 25mL aseptically.

[0031] The medium is sterilized at 121°C for 20 minutes; culture temperature: 35-37°C, culture condition: anaerobic culture.

[0032] 1.1.3 Experimental method

[0033] Under aseptic conditions, take 10 grams of Jiannanchun Laojiao Mud in a 250mL triangular flask filled with 90mL of sterile water and glass beads, place it on a bed and shake it at room temperature for 30 minutes. Treat it in an 80-degree water bath for 10 minutes, take the bacterial solution and dilute it to 10 times in a 10-fold gradient -6 , take 1mL of the b...

Embodiment 2

[0034] Caproic acid production condition optimization experiment of the bacterial strain of embodiment 2

[0035] 1.1 Optimization of fermentation conditions for caproic acid production by strains

[0036] Select temperature, fermentation days, sodium acetate and ethanol concentration as four factors and three levels to design an orthogonal experiment, and the experimental results are shown in Table 1.

[0037] Orthogonal experiment results showed that the concentration of ethanol had a greater effect on caproic acid production, followed by the concentration of sodium acetate, and the effect of temperature and fermentation time was relatively small. The optimum conditions for caproic acid production are: temperature 37°C, incubation time 15 days, ethanol concentration 2%, sodium acetate concentration 1%.

[0038] The caproic acid production test was carried out on 8 single strains metabolizing caproic acid using the optimized fermentation culture conditions. The caproic acid ...

Embodiment 3

[0042] The identification of embodiment 3 bacterial strains

[0043] 16SrDNA Sequence Analysis Method of 1F6 Strain

[0044] 1.1 DNA extraction

[0045] The cells were collected by centrifugation and dissolved in 5 mL of extraction buffer (100 mM Tris Cl, 100 mM EDTA-Na 2 , 200mM NaCl, 2%CTAB, pH8.0), shake at 37°C for 45min. Add 0.75mL20%SDS, 65 ℃ water bath 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. Centrifuge at 12,000 rpm at 4°C for 20 min to collect the precipitate, rinse twice with 70% ethanol, dry and dissolve in 50 μL TE.

[0046] 1.2PCR amplification

[0047]Using total DNA as a template, bacterial universal primers EU27F (AGAGTTTGATCCTGGCTCAG) and 1492R (GGTTACCTTGTTACGACTT) were used a...

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Abstract

The invention belongs to the technical field of wine brewing, and relates to clostridium sporogenes and the purpose of the clostridium sporogenes. According to the technical problem to be solved, a new bacterial strain with strong caproic acid producing capacity is provided. According to the technical scheme, the bacterial strain of the clostridium sporogenes is preserved in the CGMCC on February 1, 2012, the preservation number is CGMCC No. 5741, the preservation address is Institute of Microbiology of Chinese Academy of Sciences, No. 3, yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the postcode is 10010. The caproic acid producing capacity of the clostridium sporogenes can reach 9000mg/L, and is the new bacterial strain with a high caproic acid yield. A new choice is provided for the caproic acid production.

Description

technical field [0001] The invention belongs to the technical field of brewing, and relates to a Clostridium sporogenes and its application. Background technique [0002] In the production of Luzhou-flavor Daqu wine, the older the fermentation cellar, the better the quality of the wine produced. This is a summary of the experience of traditional crafts. In order to reveal the mystery of the excellent quality of old pits, my country has carried out research on the relationship between Luzhou-flavor liquor and pit mud microorganisms since the 1960s. Sexual Clostridia, which participate in the aroma production of Luzhou-flavor liquor fermentation. Ethyl caproate is the main aroma component of Luzhou-flavor liquor. Undoubtedly, the strain that metabolizes to produce caproic acid is the main functional bacteria that ferment and produce aroma in Laojiao. [0003] After the 1970s, liquor technicians made a lot of reports on the isolation, cultivation and application of caproic aci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/40C12G3/02C12R1/145
Inventor 樊科权唐清兰徐姿静徐占成
Owner 四川绵竹剑南春酒厂有限公司
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