Bacillus subtilis and culture method and application thereof

A Bacillus subtilis, culture method technology, applied in microorganism-based methods, biochemical equipment and methods, bacteria and other directions, can solve the problems of insufficient release of beneficial effects, poor degradation selectivity of strains, insufficient contact of strains, etc., to achieve excellent Straw adhesion, no laccase activity, mild treatment effect

Active Publication Date: 2019-10-15
INST OF AGRI RESOURCES & ENVIRONMENT SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the production of porous straw modified materials is still dominated by physical and chemical methods such as high temperature and high pressure treatment and chemical reagent treatment. Although these methods have high efficiency, they often consume a lot of energy and easily cause secondary pollution; Although the microbial method for straw modification is more environmentally friendly, the degradation selectivity of the strains is often poor, which can easily cause serious damage to the original structure of the straw; Insufficient contact, and the physiological effects of the strains are more due to the overall function of the cells rather than relying solely on extracellular enzymes, resulting in the inability to fully release the beneficial effects, low processing efficiency, and long processing cycles and other problems that still need to be overcome

Method used

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  • Bacillus subtilis and culture method and application thereof
  • Bacillus subtilis and culture method and application thereof
  • Bacillus subtilis and culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The screening process of Bacillus subtilis (Bacillus subtilis) ASD05 in the present invention is as follows:

[0048] (1) Primary screening

[0049] ①Collect fresh vermicompost from an earthworm farm in Shandong Province, put it into a sterile bag, store it in an ice bag and transport it back to the laboratory.

[0050] ② In order to eliminate the interference of unnecessary bacteria such as Escherichia coli and Enterococcus faecalis in vermicompost as much as possible, the samples were treated in a 70°C incubator for 2 hours and then taken out.

[0051] ③Crush the vermicompost samples under aseptic conditions, carry out gradient dilution with sterile distilled water, and select 10 -5 ~10 -8 Gradient dilutions were spread on the plates of screening medium 1, and cultured at 37°C for 24-48 hours until complete and clear colonies were formed;

[0052] Screening medium 1, the components are as follows:

[0053] Peptone 10g / L, yeast powder 5g / L, NaCl 10g / L, xylan 5g / L, ...

Embodiment 2

[0071] Identification of Bacillus subtilis ASD05 described in Example 1

[0072] (1) Strain morphology and physiological and biochemical identification:

[0073] Bacillus subtilis (Bacillus subtilis) ASD05 is cultured on a nutrient agar medium plate at 37°C for 24 hours to form white, round colonies with a diameter of 5-15mm and a thickness of 0.5-1mm. The surface of the colonies is rough and irregular. Wrinkled, with burrs on the edge; Gram staining is positive, and the shape of the bacterium under the microscope is short rod-shaped, with a size of 0.8-1.2μm×2-4μm, with endophytic spores, and the bacterium does not expand after the formation of spores, and the spores Located in the middle of the cell, the bacteria can move, see the colony and cell morphology of the strain figure 1 and figure 2 .

[0074] The main physiological and biochemical characteristics of Bacillus subtilis (Bacillus subtilis) ASD05, the test results are: contact enzyme experiment, glucose acid produ...

Embodiment 3

[0082] The cultivation method of Bacillus subtilis (Bacillus subtilis) ASD05 described in embodiment 1

[0083] (1) Take the activated Bacillus subtilis ASD05, inoculate it into a shaker flask equipped with a seed medium, and culture it on a shaker at 37° C. and 220 rpm for 24 hours to obtain liquid seeds;

[0084] The components of the seed culture medium are as follows: peptone 10g / L, yeast powder 5g / L, NaCl 10g / L, glucose 5g / L, and balance water;

[0085] The activated Bacillus subtilis (Bacillus subtilis) ASD05 is to inoculate Bacillus subtilis (Bacillus subtilis) ASD05 into the activation medium, and activate the culture at 37° C. for 24 hours; the activation medium is LB solid medium.

[0086] (2) Take the liquid seeds prepared in step (1), inoculate them in a shake flask equipped with an expansion medium at a ratio of 0.5% by volume, and culture them on a shaking table at 37° C. and 220 rpm for 36 hours to obtain the bacterial liquid;

[0087] The components of the exp...

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Abstract

The invention relates to bacillus subtilis and a culture method and application thereof. The bacillus subtilis ASD05 is preserved in the General Microbiology Center of the China Committee for CultureCollection of Microorganisms on May 15, 2019, the preservation address is No.3, No.1 Courtyard, Beichen West Road, Chaoyang District, Beijing City, and the preservation number is CGMCC No.17807. The invention further relates to the culture method and application of the bacillus subtilis ASD05. It is found for the first time that the bacillus subtilis ASD05 has the effect of adhering microorganismsrich in straw degradation effect to the surface of straw, and meanwhile has the capacity of high yield of xylanase and pectinase, cellulase activity is low, and laccase activity is avoided.

Description

technical field [0001] The invention relates to a strain of Bacillus subtilis and its cultivation method and application, belonging to the technical field of microbes. Background technique [0002] Straw is an important biomass resource. The total amount of straw resources in my country is huge. According to data, as of 2018, the annual output of various straw resources in my country has exceeded 1 billion tons. The resource utilization of straw not only has huge economic value, but also has important environmental protection significance. At present, the resource utilization methods of straw mainly include returning to the field, feeding, composting, biogasification, biochemical and other methods. "Green waters and lush mountains are golden mountains and silver mountains." No matter what treatment method is adopted, while stimulating the utilization potential of straw resources, it is necessary to reduce and eliminate environmental pollution factors, especially to avoid se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20D21C5/00C12R1/125
CPCC12N1/20C12N1/205C12R2001/125D21C5/005
Inventor 付龙云李彦王艳芹赵自超袁长波井永苹薄录吉
Owner INST OF AGRI RESOURCES & ENVIRONMENT SHANDONG ACADEMY OF AGRI SCI
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