Clostridium butyricum agent with strong butyric acid resistance and stress resistance and application thereof

A technology of Clostridium butyricum and bacteria agent, applied in the field of microbial fermentation, can solve the problems affecting the application of Clostridium butyricum and low fermentation level, achieve strong application value, improve fermentation level, and increase daily weight gain

Active Publication Date: 2018-04-20
湖北绿雪生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because its low level of fermentation seriously affects the application of Clostridium butyricum, how to reduce the production cost of Clostridium butyricum is imminent, and screening high-yield and stress-resistant Clostridium butyricum strains through mutation breeding is Clostridium butyricum Main channels of product promotion

Method used

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  • Clostridium butyricum agent with strong butyric acid resistance and stress resistance and application thereof
  • Clostridium butyricum agent with strong butyric acid resistance and stress resistance and application thereof
  • Clostridium butyricum agent with strong butyric acid resistance and stress resistance and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The mutagenesis cultivation of embodiment 1, LXKJYB-1

[0038] (1) Suspension preparation:

[0039] Activate the Clostridium butyricum LXKJ-1 strain preserved in the glycerol tube, pick a single colony and inoculate it in the RCM liquid medium, and cultivate it for 8-10 hours according to the growth curve, that is, collect the bacterial liquid in the logarithmic phase for centrifugation, and collect Bacteria were washed two to three times with normal saline, and the mutagenesis suspension was prepared for mutagenesis. The concentration of bacteria in the mutagenesis solution was 2×10 6 cfu / ml. Growth curve see figure 1 .

[0040] (2) Make a lethal curve

[0041] The dose or time of mutagenesis treatment not only affects the lethal rate, but also can significantly affect the mutation efficiency. Effective screening in an appropriate range of mutation efficiency is the key to strain selection. In this study, the starting strains were first subjected to plasma jet treat...

Embodiment 2

[0058] Embodiment 2, the impact of different fermentation media on the fermentation level of butyric acid-resistant high-yield strain LXKJYB-1

[0059] Put the LXKJYB-1 frozen in the glycerol tube into the activation medium, and culture it statically at 37°C for 18 hours; the activated Clostridium butyricum LXKJYB-1 was inoculated into the seed medium with an inoculation amount of 5%, and anaerobically cultured at 37°C. Oxygen cultivation for 24 hours to obtain seed solution.

[0060] Fermentation medium 1: 2% ammonium sulfate, 2% corn flour, 2% yeast extract, 1% urea, 0.5% dipotassium hydrogen phosphate, pH 6.5-7.0, 0.5% calcium carbonate; dissolved in deionized water, stirred with a glass rod Mix evenly, sterilize after subpackaging, and sterilize at 121°C for 20 minutes;

[0061] Fermentation medium 2: 4% ammonium sulfate, 4% corn flour, 4% yeast extract, 2% urea, 1.5% dipotassium hydrogen phosphate, pH 6.5-7.0, 0.7% calcium carbonate; dissolved in deionized water, stirred...

Embodiment 3

[0067] Embodiment 3, the cultivation of Clostridium butyricum (Clostridium butyricum) LXKJYB-1

[0068] (1) Activation and seed cultivation

[0069] The LXKJYB-1 and LXKJ-1 strains frozen in glycerol tubes were respectively connected to the activation medium, and incubated anaerobically at 37°C for 18 hours; the activated Clostridium butyricum LXKJYB-1 and LXKJ-1 were respectively inoculated into the seed culture base, the inoculum size was 5%, and anaerobic culture was carried out at 37°C for 24 hours to obtain the seed solution.

[0070] (2) Primary seed cultivation

[0071] Inoculate the LXKJYB-1 and LXKJ-1 seed liquids activated in step (1) into the primary seed medium at 5% inoculum, anaerobically, at 37°C, and culture for 12 hours to amplify to obtain the primary seed liquid. Complete the comparison between the number of viable bacteria and the number of spores, as shown in Table 4.

[0072] (3) Secondary seed cultivation

[0073] Insert the primary seed liquid of Cl...

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Abstract

The invention belongs to the technical field of microbial fermentation, and in particular, relates to a clostridium butyricum inoculum with strong butyric acid resistance and stress resistance and anapplication thereof. Clostridium butyricum is specifically clostridium butyricum LXKJYB-1, and has the preservation number of CCTCC NO:M 2017485. The strain LXKJYB-1 has the characteristics of high fermentation level, high organic acid yield, inhibition of a variety of intestinal pathogens, and high tolerance capacity to gastrointestinal fluid, cholate and damp heat. The strain and the inoculum prepared therefrom are suitably applied for livestock and poultry breeding, increases the efficiency of breeding, and produces no-resistance livestock and poultry products.

Description

Technical field: [0001] The invention belongs to the technical field of microbial fermentation, and in particular relates to a butyric acid-resistant and stress-resistant Clostridium butyricum strain and its application. Background technique: [0002] Clostridium butyricum is a completely anaerobic, Gram-positive bacillus that is a normal flora of the intestinal tract of humans and animals and is also present in soil, cheese, and natural yoghurt; flagellated, motile; spores eccentric or Subterminal (endophytic), round or oval, without spores and appendages. The spore structure of Clostridium butyricum can resist the harsh environment of the gastrointestinal tract of animals, and can withstand high temperature and high humidity in the process of feed processing; Clostridium butyricum is only sensitive to a few antibiotics such as novobiocin, vancomycin and tetracycline, It is highly resistant to many other antibiotics and has a good application prospect. [0003] Clostridiu...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K50/75A23K10/18C12R1/145
CPCA23K10/18A23K50/75C12N1/20C12N1/205C12R2001/145
Inventor 王丽张仁文谢丽静王海宽丁立张震宇
Owner 湖北绿雪生物科技有限公司
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