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Bacillus subtilis bred by space mutation breeding technology and application of Bacillus subtilis

A Bacillus subtilis, aerospace breeding technology, applied in the biological field, can solve the problems of poor tolerance such as acid resistance, affecting feeding effect, affecting product enzyme activity, etc., so as to reduce feed cost, improve feed conversion rate, and improve digestion and absorption. Effect

Active Publication Date: 2013-01-16
山东宝来利来生物工程股份有限公司
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The research and development of feed enzyme preparations have been highly valued by the feed industry and aquaculture industry. The enzyme preparations that have been developed so far include cellulase, α-amylase, acid protease, neutral protease, phytic plum, xylan There are many kinds of enzymes, dextranases, etc., but due to the shortcomings of such enzyme preparations such as poor high temperature resistance, acid resistance, and short storage time, the enzyme activity of the product is affected, thus affecting this type of product. The feeding effect of the animal greatly limits the promotion and application of feed enzyme preparations, and it is urgent to develop a new type of animal microecological preparation

Method used

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  • Bacillus subtilis bred by space mutation breeding technology and application of Bacillus subtilis
  • Bacillus subtilis bred by space mutation breeding technology and application of Bacillus subtilis
  • Bacillus subtilis bred by space mutation breeding technology and application of Bacillus subtilis

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment one, the screening of strain

[0030] 1. Materials and methods

[0031] 1.1 Samples: soil, hay, animal intestinal contents, feces of cattle, sheep, geese and other animals, and cattle rumen contents.

[0032] 1.2 Screening medium:

[0033] Screening medium for amylase-producing strains: 1.0% corn starch, 0.3% beef extract, 0.5% peptone, 2.0% agar powder, pH 7.2-7.4, sterilized at 121°C for 30 minutes.

[0034] Screening medium for neutral protease-producing strains: soybean protein powder 1.0%, beef extract 0.5%, dipotassium hydrogen phosphate 0.05%, magnesium sulfate 0.05%, calcium chloride 0.05%, Tween-800.1%, agar powder 2.0%, pH7.2~7.4, 121℃, sterilize for 30min.

[0035] Screening medium for cellulase-producing strains: carboxymethyl cellulose 1%, peptone 0.5%, dipotassium hydrogen phosphate 0.05%, magnesium sulfate 0.02%, calcium chloride 0.01%, ferric chloride 0.02%, agar powder 2.0% , pH 7.2-7.4, sterilized at 121°C for 30 minutes.

[0036] Scree...

Embodiment 2

[0050] Embodiment two, identification and safety test of bacterial classification

[0051] 1. Materials and methods

[0052] 11 strains: The microorganisms isolated from cow dung and goose dung samples were screened out after primary screening and secondary screening, and a total of 2 strains with high enzyme activity such as neutral protease were screened out, and the numbers were N and E respectively.

[0053] 1.2 Routine biochemical identification of bacteria: Unless otherwise specified, tests for general morphology, physiological and biochemical properties were carried out with reference to "Bergey's Bacterial Identification Manual" (9th edition) and "Microbiology Experiment Manual".

[0054] 1.3 Safety test: different concentrations (1×10 8 cfu / mL, 1×10 9 cfu / mL, 1×10 10 cfu / mL, 1×10 11 cfu / mL) of the N bacterial strain and the fermented liquid of the E bacterial strain were injected intraperitoneally to mice, 10 in each group, and observed continuously for 1 week to ...

Embodiment 3

[0061] Embodiment three, the selection and breeding of N strain mutagenization bacterial classification

[0062] 1. Materials and methods

[0063] 11 strains: Bacillus subtilis N strain

[0064] 1.2 Medium

[0065] 1.2.1 Liquid seed medium

[0066] 10g peptone, 5g sodium chloride, 5g beef extract, 1000mL water, pH7.2. Sterilize at 121°C for 30min.

[0067] 1.2.2 Liquid fermentation medium

[0068] Glucose 5g, peptone 10g, sodium chloride 5g, beef extract 5g, water 1000mL, pH7.2.

[0069] Capacity: 100mL in a 500mL Erlenmeyer bottle, sterilized at 121°C for 30min.

[0070] 1.3 Enzyme activity determination method and apparatus are the same as in Example 1.

[0071] 1.4 Mutagenesis method:

[0072] The 18th returnable science and technology satellite launched by China's Jiuquan Satellite, after 18 days of orbiting, in the special environment of space microgravity, strong radiation, high-energy particles, alternating magnetic field, etc., the genetic mutation of the bacter...

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Abstract

The invention discloses Bacillus subtilis bred by a space mutation breeding technology. The strain is named Bacillus subtilis N9-1-35 and was collected in China Center for Type Culture Collection on August 31, 2011, and the collection number is CCTCC M2011301. The test shows that the Bacillus subtilis is an excellent probiotic strain. After entering intestinal tracts of animals, the Bacillus subtilis achieves the effects of a probiotic and can also generate a plurality of kinds of digestive enzymes in the growth and reproduction process of the animals, so that the digestion and absorption of the animals on feeds are improved, the conversion rate of the feeds is improved, and the cost of the feeds is reduced; and the Bacillus subtilis can be used for producing enzyme-producing probiotics, and is a strain which has excellent research and development value. The invention also discloses an enzyme-producing probiotic. The enzyme-producing probiotic consists of Bacillus subtilis powder, Streptococcus lactis powder, Lactobacillus acidophilus powder and corn starch.

Description

technical field [0001] The invention relates to a bacillus subtilis bred by space breeding technology and its application, belonging to the field of biotechnology. Background technique [0002] In the 21st century, people have higher and higher requirements for food safety and ecological environment, and the feed industry, which is at the forefront of the aquaculture industry, has generally attracted people's attention. The "1996-2020 China Feed Industry Development Strategy Research" promulgated by the National Feed Industry Office has clearly stated that "it is very urgent to replace the abuse of chemical synthetic substances such as feed antibiotics with green environmental protection products". The best way to replace these pollutants is to use biological technology. Microecological preparation is a modern high-tech product of biotechnology. It is a functional biological feed additive that organically combines probiotics, enzyme preparations, oligosaccharides, and biolo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K1/16C12R1/125C12R1/46C12R1/23A23K10/18
Inventor 单宝龙谷巍徐海燕翟延庆
Owner 山东宝来利来生物工程股份有限公司
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