Application of anti-mildew bacillus strain and antimicrobial product thereof in corn storage

A Bacillus and Bacillus subtilis technology, applied in the biological field, can solve the problem of no Bacillus antibacterial products, etc., and achieve the effects of ensuring food safety, ensuring food quality, and improving food quality

Active Publication Date: 2014-08-27
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is still no report of Bacillus antimicrobial products used in corn mildew prevention

Method used

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  • Application of anti-mildew bacillus strain and antimicrobial product thereof in corn storage
  • Application of anti-mildew bacillus strain and antimicrobial product thereof in corn storage
  • Application of anti-mildew bacillus strain and antimicrobial product thereof in corn storage

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Screening of anti-mold Bacillus subtilis LPL-NM45

[0058] 1. Isolation of Aspergillus niger, Penicillium oxalicum and Fusarium moniliforme

[0059] Isolation of Aspergillus niger, Penicillium oxalicum and Fusarium moniliforme from corn samples. Separation method: Weigh 25g of corn sample, add it to 225mL sterilized normal saline, oscillate fully to make a spore suspension, dilute 10-1000 times with sterile normal saline, draw 0.1mL into a petri dish, then pour it into Cool to 45°C PDA medium, culture at 28°C for 3-5 days, pick a single colony plate, streak and purify, and identify, isolate Aspergillus niger, Penicillium oxalicum, and Fusarium moniliforme.

[0060] 2. Use LB medium to isolate Bacillus from different samples

[0061] In order to ensure the diversity and safety of the obtained strains, try to select samples from relatively safe foods and biological environments that may contain more bacillus. The samples are Xinjiang roasted naan, thick soup ...

Embodiment 2

[0078] Example 2 Identification of producing antibacterial substance bacillus LPL-NM45

[0079] 1. Morphological identification

[0080] Pick a small amount of the target strain Bacillus LPL-NM45 and streak it on the LB plate, culture it at 28°C for 20 hours, observe the single colony shape, shape, size, edge, surface, raised shape, transparency, colony and color of the culture medium, etc., continue After 24h and 48h of culture, microscopic examination was performed to record the morphology of the bacteria.

[0081] After being cultured in LB medium for 24 hours, the colony shape of the strain LPL-NM45 was regular, the surface of the colony was wrinkled, the edges were neat, opaque, and wet and sticky, such as Figure 4 shown. Under the microscope, the slender rod-shaped, spores are oval in shape, enlarged, and proximally grown, such as Figure 5 shown.

[0082] 2. Physiological and biochemical identification

[0083] Refer to the "Common Bacteria System Identification M...

Embodiment 3

[0099] Example 3 The antimicrobial product of Bacillus subtilis LPL-NM45 controls the mold during high moisture corn storage

[0100] 1. Inhibition and effect of antibacterial products of Bacillus subtilis LPL-NM45 on corn mold and its spores

[0101] 1.1 Extraction of antibacterial products of Bacillus subtilis LPL-NM45

[0102] Inoculate Bacillus subtilis LPL-NM45 in LB liquid medium, culture at 140rpm at 37°C for 48h to obtain a fermentation broth, centrifuge the fermentation broth at 10,000rpm for 12min to obtain a fermentation supernatant, add ammonium sulfate solid powder to the fermentation supernatant to Saturation is 65%, 4°C, 200rmp after stirring for 1h, place in 4°C refrigerator overnight, centrifuge at 10000rpm for 12min, discard supernatant, and redissolve the precipitate in phosphate buffer (25mM pH7.0) 1 / 10 of the volume of fermentation broth In the middle, 3.5kDa dialysis bag is dialyzed, and after the dialysis ends, the antibacterial activity of the solution...

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Abstract

The invention provides an anti-mildew bacillus strain, namely Bacillus subtilis LPL-NM45 of which the collection number is CGMCC No.7762. The invention also provides a method for extracting an antimicrobial product of the Bacillus subtilis LPL-NM45. The Bacillus subtilis LPL-NM45 and the antimicrobial product thereof have strong inhibiting effect on growth of mildew and germination of spores in grain storage (especially corn storage). In case of the identical spraying dosage, the Bacillus subtilis LPL-NM45 and the antimicrobial product thereof can more effectively inhibit the growth of mildew in the storage process of high-water-content corns in comparison with the conventional chemical preservative used in grains, and guarantee the safety of the grains, thus having favorable prospects of being used as mildew inhibitors for grains.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a mold-resistant Bacillus strain, the inhibitory effect of an antibacterial product produced by the strain on mold growth and spore germination, and its anti-mold application in corn storage. Background technique [0002] Bacillus exists widely in nature. It is a thermophilic, aerobic and spore-forming Gram-positive rod-shaped bacteria that can produce a variety of antibacterial substances and enzymes. It has strong anti-mold and disease-resistant effects, and is extremely Easy to isolate and cultivate, it is one of the main antibacterial protein producing bacteria. [0003] There are two ways for Bacillus to produce antibacterial substances: ribosomal and non-ribosomal synthesis. Antibacterial substances produced by the ribosomal pathway include bacteriocins, bacteriocinoids, enzymes, and active proteins. Antibacterial substances produced by non-ribosomal pathways include lipopept...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P1/04A23B9/28A23B9/24C12R1/125
Inventor 李平兰丛苑杜奕君韩玉竹卢彦廷梁志宏
Owner CHINA AGRI UNIV
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