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Bacillus velezensis strain for soybean meal fermentation

A technology of Bacillus Velez and Bacillus, applied in the direction of bacteria, applications, microorganisms, etc., can solve problems such as single process and detection technology, affecting protein quality, and large differences in protein solubility and small peptide content

Active Publication Date: 2020-10-27
CHONGQING ACAD OF ANIMAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, at present, most of the strains used in the existing soybean meal fermentation technology on the market are non-specific fermentation strains, and there are large differences in the fermentation process, resulting in the protein solubility of fermented soybean meal (ranging from 30% to 80%), small The peptide content varies greatly, which affects the quality of the protein, and is limited by the single process and detection technology. Not all fermented soybean meal on the market is a real low-antigen protein product

Method used

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  • Bacillus velezensis strain for soybean meal fermentation
  • Bacillus velezensis strain for soybean meal fermentation
  • Bacillus velezensis strain for soybean meal fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0033] The isolation and screening of Bacillus veleisi strain DP-2, the specific steps are:

[0034] a. Isolation and purification of Bacillus in Douchi

[0035] Take 5g of Douchi sample into a 250mL Erlenmeyer flask filled with 45mL sterilized normal saline (add 10% LB culture solution), shake and incubate for 30min, place the Erlenmeyer flask containing the sample solution in a water bath at 80°C for 20min, The sample solution was diluted 10 times with sterile physiological saline, and the appropriate dilution gradient suspension was spread on the plate of LB medium, 3 plates for each dilution, and cultured upside down at 37°C. After the colonies grow out, pick the strains with the morphological characteristics of Bacillus colonies, and separate and purify them by streaking.

[0036] b. Primary screening

[0037] The isolated and purified strains were planted on the screening plate with a sterilized toothpick, cultured upside down at 37°C for 18-24 hours, and the strains t...

Embodiment 2

[0063] Identification and Growth Curve Measurement of DP-2 Strain

[0064] Bacterial strain DP-2 grows rapidly on the LB plate, and the colonies are milky white, opaque, with wrinkled edges, raised central part, easy to stir up, and sticky (such as figure 2 shown), and the shape of the bacteria was rod-shaped observed under the optical microscope (such as image 3 shown), morphologically identified as subspecies of Bacillus subtilis; identified by molecular biology as Bacillus Velez (such as Figure 4 shown); the growth and development curve of the strain is as follows Figure 5 shown.

[0065] Molecular biology identification is specifically as follows: use 16S rDNA universal primers: 338F (5′-ACTCCTACGGGAGGCAGCA-3′) and 806R (5′-GGACTACHVGGGTWTCTAAT-3′) to amplify the 16S rDNA of the target strain, and carry out the purified PCR product Sequencing, the sequencing results were Blast compared in the genbank nucleic acid sequence database to search for homologous sequences;...

Embodiment 3

[0068] Detect the protease-producing properties of the DP-2 strain. The specific steps are: after the DP-2 bacterial liquid is centrifuged at 3000r / min for 10 minutes to remove the bacteria, it is dialyzed with a 2KD dialysis bag for 24 hours (change the water 2-3 times in the middle), and the dialysis bag is taken out. The inner solution is freeze-dried (pre-freezing at -40°C for 540min, vacuuming at -40°C to 1500MT for 120min, first drying at 10°C at a vacuum of 200MT for 740min, at 25°C at a vacuum of 200MT for 1020min, and secondary drying at 20°C at a vacuum of 1020min 300MT dried for 120min and cooled to 0°C) and weighed to measure the activity of each protease in the lyophilized powder; the results are shown in Table 3;

[0069] Among them, the protease activity is detected according to the corresponding detection method of "GB / T 23527-2009 Protease Preparation".

[0070] Table 3 DP-2 bacterial liquid protease detection results

[0071] Enzyme activity Neut...

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Abstract

The invention belongs to the field of bacteria / livestock microorganisms, and particularly relates to a bacillus velezensis strain for soybean meal fermentation. The strain is bacillus velezensis DP-2with a preservation number of CGMCC No.17235. The bacillus velezensis strain DP-2 can efficiently secrete neutral protease and alkaline protease, and has a good degradation effect on anti-nutrient substances such as glycinin and beta-conglycinin in soybean meal; and the degradation rate of the glycinin can reach 81%-96%, the degradation rate of the beta-conglycinin can reach 50%-67%, acid-solubleprotein is increased by 5-6 times, the content of nutrient substances in fermented soybean meal can be remarkably increased, the nutrient utilization rate is remarkably increased, and therefore the nutritional value of the soybean meal is improved.

Description

technical field [0001] The invention belongs to the field of bacteria / animal husbandry microorganisms, and in particular relates to the application of a Bacillus velei strain for soybean meal fermentation in feed raw material soybean meal. Background technique [0002] Soybean meal is a by-product obtained from soybean oil extraction or pre-press extraction. It has high protein content and various nutrients. ("Functional properties of soybean meal and its research and development status", Chang Qing et al., Food Research and Development, 2010, Volume 31, No. 10, pp. 217-220, published on October 31, 2010; Research on the Evaluation of Nutritional Value", Peng Huicai, Guangxi University Master's Degree Thesis, 2008, p. 1, published on November 06, 2008). However, soybean meal contains a variety of anti-nutritional factors such as glycinin, β-conglycinin (accounting for more than 70% of total soybean protein), trypsin inhibitors, bad oligosaccharides, etc., which not only red...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K10/37A23K10/12C12R1/07
CPCA23K10/12A23K10/37C12N1/20C12N1/205C12R2001/07Y02P60/87
Inventor 刘志云官小风周晓容杨飞云姚焰础
Owner CHONGQING ACAD OF ANIMAL SCI
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