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Bacillus pumilus viable preparation and solid fermentation method and applications thereof

A Bacillus pumilus and solid fermentation technology, applied in the field of microbial fermentation, can solve the problems of low treatment yield, low fermentation bacteria amount, poor heat resistance of spores, etc., and achieve the effects of short fermentation time, high bacteria amount and low cost

Inactive Publication Date: 2015-02-25
QINGDAO GENYUAN BIOLOGICAL TECH GRP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been used for liquid fermentation, the amount of fermentation bacteria is low, and the heat resistance of spores is poor, and the yield of spray drying treatment in the later stage is low

Method used

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  • Bacillus pumilus viable preparation and solid fermentation method and applications thereof
  • Bacillus pumilus viable preparation and solid fermentation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Screening of antagonistic bacteria

[0028] 1. Isolation and screening of Bacillus pumilus S3

[0029] From May 2013 to September 2013, water samples, sediment, and intestinal feces of prawns were collected from many places along the coast of Jiaozhou. The bottom mud and shrimp intestinal feces samples were diluted 10 times in sterile normal saline, fully ground, and the supernatant was taken as the sample solution; the water sample was directly used as the sample solution; the sample solution was inoculated in 2216E liquid medium for enrichment at 30°C After culturing for 48 h, it was diluted and spread on 2216E plates to isolate and purify the strains. A total of 26 pure strains were screened out.

[0030] The purified single strains were screened for antagonistic bacteria by plate spotting method. Using Vibrio parahaemolyticus as indicator bacteria, Vibrio parahaemolyticus was inoculated in 2216E liquid medium for 18 h and diluted to 10 5 ~10 6 For CFU...

Embodiment 2

[0037] The solid fermentation method of the bacillus pumilus live bacteria preparation of the present invention specifically comprises the following steps:

[0038] 1. Activate the selected Bacillus pumilus S3;

[0039] 2. Place the activated Bacillus pumilus S3 in the seed culture solution for resuscitation and subculture to obtain a seed solution. The liquid seed medium was prepared according to the following formula: 30 g of enzymatically hydrolyzed soybean meal, 10 g of cornmeal, 50 g of glucose, 5 g of NaCl, 1 L of water, and the pH value was adjusted to 7.0. The prepared above-mentioned liquid seed medium was sterilized by high-pressure steam at 121°C for 20 minutes. Take the activated bacteria in 150ml liquid seed medium, inoculum volume ratio is 2% (V / V), culture at 37°C, 180rpm, take samples regularly to measure OD660nm, from figure 1 It can be seen from the figure that S3 grows faster and enters a stable period at 12 hours, and gradually begins to form spores in t...

Embodiment 3

[0043] Take 1 g of the Bacillus pumilus preparation prepared above and soak it in 100 ml of water for 6 hours, centrifuge to get the supernatant to measure its inhibitory effect on Vibrio parahaemolyticus. Dilute Vibrio parahaemolyticus solution to 10 5 -10 6 CFU / ml, take 0.1 ml and coat 2216E plate, place 3 sterile Oxford cups on the plate equidistantly, add 200 ul Bacillus pumilus S3 liquid fermentation broth, Bacillus pumilus S3 solid fermentation soaking solution respectively, and incubate at 25°C for 24 hours Observe and measure the diameter of the inhibition zone. Such as figure 2 As shown, the solid fermentation of Bacillus pumilus S3 has obvious antibacterial effect (d=1.5cm), which is better than that of liquid fermentation (d=1.1cm).

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Abstract

The invention discloses a bacillus pumilus viable preparation and a solid fermentation method and applications thereof, belonging to the field of microbial technology. The collection number of the selected bacillus pumilus is CGMCCNo.9161. The high-density fermentation method of bacillus pumilus comprises the following steps: selection of strains, culturing of seeds, solid fermentation, drying of fermentation products, and packaging of finished product. By adopting bagasse and coarse bran as active ingredients of the culture medium, the cost is low, the strain amount is high, the number of effective viable bacillus pumilus can achieve 1*10<11>CFU / g above; through adding sodium citrate, the spore formation time can be obviously reduced, the spore maturate rate can be improved, and the shelf life of products can be prolonged.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and in particular relates to a live bacillus pumilus preparation and a solid fermentation method and application thereof. Background technique [0002] Bacillus pumilus ( Bacillus pumilus ) is rod-shaped, with rounded ends, single or arranged in short chains, Gram-positive, oval, meso-terminal or sub-terminal; it belongs to feed microbial strains, suitable for pigs, poultry, ruminants, and fish , Shrimp and other aquatic animals. [0003] A strain of Bacillus pumilus involved in the present invention can produce a variety of exogenous digestive enzymes, such as xylanase, cellulase, etc., and can also secrete antibacterial substances, which are effective against Escherichia coli, Aeromonas hydrophila, Hemolytic Vibrio and other pathogenic bacteria have inhibitory effect. Liquid fermentation has been adopted for it, the amount of fermentation bacteria is low, and the heat resistance of sp...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/02A01P1/00C12R1/07
CPCA01N63/10C12N1/20
Inventor 王娇李慧芬梁晶晶
Owner QINGDAO GENYUAN BIOLOGICAL TECH GRP
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