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Temperature-resistant and acid-alkali-resistant strain, screening method, microbial inoculum and application

A technology of acid and alkali resistance and bacterial strains, applied in the field of applied bacterial strains, can solve the problems of few and few disclosure of ultra-high temperature thermophilic bacteria agents.

Pending Publication Date: 2022-04-12
HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, at present, the research on microbial fermentation inoculum at home and abroad focuses on the development of microbial inoculum with room temperature and mesophilic strains such as Bacillus and Aspergillus, and there are not many reports on ultra-high temperature thermophilic inoculum for composting, especially for composting. Ultra-high temperature thermophilic bacteria products with good adaptability and functionality are rarely disclosed

Method used

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  • Temperature-resistant and acid-alkali-resistant strain, screening method, microbial inoculum and application
  • Temperature-resistant and acid-alkali-resistant strain, screening method, microbial inoculum and application
  • Temperature-resistant and acid-alkali-resistant strain, screening method, microbial inoculum and application

Examples

Experimental program
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Embodiment 1

[0049] The present embodiment is the screening method of Bacillus urea L2, which comprises the following steps:

[0050] (1) Isolation of strains

[0051] Composted product samples from the food waste composting plant in Shijiazhuang City, Hebei Province were passed through LB solid medium (formulation: tryptone 10g / L, yeast extract 5g / L, NaCl 10g / L) to isolate and obtain 21 bacterial strains, which were plate-passaged 5 times to obtain stable genetic monoclonal strains, and stored at -80°C.

[0052] (2) Preliminary screening of strains with high temperature resistance and high activity cellulase

[0053] The isolated 21 monoclonal strains were treated with cellulose Congo red medium (recipe: sodium nitrate 1.0g, disodium hydrogen phosphate 1.2g, potassium dihydrogen phosphate 0.9g, magnesium sulfate 0.5g, potassium chloride 0.5g, yeast leaching 0.5g powder, 0.5g acid hydrolyzed casein, 0.2g Congo red, 5.0g cellulose powder, 15.0g agar, pH 7.0±0.1, 1000ml distilled water, a...

Embodiment 2

[0065] This embodiment relates to the identification of physiological characteristics of Bacillus urea L2, including the identification of high temperature resistance and acid and alkali resistance, the method is as follows:

[0066] (1) Streak-inoculate Bacillus geourea L2 stored at -80°C on LB solid plates (see above for recipe), and inoculate them at 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, and 70°C, respectively. , 75°C, 85°C and 90°C for 3 days, and observe the growth status of bacterial strain L2 under different temperature conditions, the results are shown in Table 2:

[0067] Table 2 Growth status of L2 at different temperatures

[0068]

[0069] Note: + indicates that it can grow (there are colonies on the solid medium), ++ grows well (the number of colonies on the solid medium is 10-50), +++ grows vigorously (the number of colonies on the solid medium is greater than 50) indivual).

[0070] It can be seen from Table 2 that Bacillus urea L2 can grow well on LB medium ...

Embodiment 3

[0077] This example relates to the identification of enzyme-producing activities of Bacillus urea L2, including producing lipase, protease and cellulase. The identification method is as follows:

[0078] (1) Verify the lipase-producing ability of the Bacillus urea L2 strain: activate the L2 strain and inoculate it into the nutrient broth medium (peptone 10g / L, beef powder leaching powder 3g / L, sodium chloride 5g / L, pH 7.2±0.2), cultured at 60°C for 24h. The activity of lipase in culture medium was determined by alkaline titration. It was determined that the activity of lipase in the culture medium was 10.2U / mL.

[0079] (2) The L2 strain was activated and inoculated into 50 mL of skimmed milk powder medium (10 g of peptone, 3 g of beef extract, 5 g of sodium chloride, 1.5 g of skim milk powder, and 1000 ml of distilled water), and cultured at 50°C for 24 hours. The protease activity in the mixture was determined by the Folin-phenol method. It was determined that the prote...

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Abstract

The invention relates to a temperature-resistant and acid-alkali-resistant bacterial strain, which is Ureibacillus terrenus, is named as Ureibacillus terrenus L2, is preserved in China General Microbiological Culture Collection Center (CGMCC) on July 30, 2021, and has a preservation number of CGMCC No.22991. The temperature-resistant and acid-alkali-resistant bacterial strain has the advantages that the temperature-resistant and acid-alkali-resistant bacterial strain can be applied to the field of agricultural production, and the production cost is low; the strain can keep higher activity at 40-90 DEG C, can survive in an environment with wider pH (4.0-11.0) range and keep higher activity, and simultaneously has remarkable cellulase activity, protease activity and lipase activity, so that the strain can be widely applied to an aerobic composting process of various agricultural solid wastes (including but not limited to livestock and poultry manure and crop straws), and can be applied to the aerobic composting of various agricultural solid wastes (including but not limited to livestock and poultry manure and crop straws). An available microbial agent is provided for efficient composting. In addition, the invention also relates to a screening method and application of the strain.

Description

technical field [0001] The invention relates to the technical field of applied bacterial strains, in particular to a temperature-resistant, acid- and alkali-resistant bacterial strain, a screening method, bacterial agents and applications. Background technique [0002] Every year, farms produce a large amount of livestock and poultry manure and sewage, and livestock and poultry manure poses a serious threat to the surrounding environment. Composting is a widely used aerobic fermentation technology and one of the treatment methods for livestock and poultry waste. Through the metabolism of aerobic microorganisms, complex macromolecular organic matter can be decomposed into small molecules, and sterilized to make organic fertilizers. Throughout the process, aerobic microorganisms continue to multiply and decompose organic matter. Thermoaerobic composting is an environmentally friendly process and a sustainable alternative to managing and recycling organic matter in order to o...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02C05F17/20C12R1/01
CPCY02W30/40
Inventor 李再兴李雪梅马骏武肖莎秦学李贵霞张凡韩学滨
Owner HEBEI UNIVERSITY OF SCIENCE AND TECHNOLOGY
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