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Bacillus subtilis H4, decomposed inoculant prepared therefrom and application of the decomposed inoculant

A technology of Bacillus subtilis and decomposing bacteria agent, which is applied in the application, preparation of organic fertilizers, bacteria, etc., can solve the problems of low compost fermentation efficiency, achieve high protease activity, rapid growth, and low environmental pollution

Active Publication Date: 2015-02-04
HANGZHOU FAMOXI BIOMEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the composting process is greatly affected by factors such as microbial species and ambient temperature, the composting fermentation efficiency is low without the artificial addition of bacterial agents

Method used

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  • Bacillus subtilis H4, decomposed inoculant prepared therefrom and application of the decomposed inoculant
  • Bacillus subtilis H4, decomposed inoculant prepared therefrom and application of the decomposed inoculant
  • Bacillus subtilis H4, decomposed inoculant prepared therefrom and application of the decomposed inoculant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 strain screening and identification

[0056] 1. Screening and identification of strain H4

[0057] (1) Culture medium and reagent formula:

[0058] The final concentration of SOC liquid medium is composed of: tryptone 20g / L, yeast extract 5g / L, NaCl 0.5g / L, KCl 2.5mmol / L, MgCl 2 0.01mol / L, glucose 0.02mol / L, solvent is deionized water, pH value 7.0.

[0059] The final concentration of the SOC solid medium is composed of: SOC liquid medium plus agar powder with a final mass concentration of 1.5%.

[0060] Preparation method of SOC liquid medium: For each liter of medium, add 20g of tryptone, 5g of yeast extract, and 0.5g of NaCl into 965ml of deionized water and shake the container to completely dissolve the solute. Add 10ml 250mmol / L KCl aqueous solution, use 5mol / L NaOH to adjust the pH to 7.0, at 15psi (1.05kg / cm 2 ) steam sterilization under high pressure for 20min. Add 5ml filter-sterilized 2mol / L MgCl to the sterilized solution 2 , cooled to 60°C or...

Embodiment 2

[0133] Embodiment 2 decomposed bacteria agent

[0134] (1) Bacillus subtilis H4 freeze-dried powder

[0135] a. Incline cultivation

[0136] Bacillus subtilis H4 was inoculated into slant LB medium and cultured at 37°C for 1 day to obtain slant cells.

[0137] b. Seed cultivation

[0138] Pick an inoculation loop strain from the slant bacteria, inoculate it into 100ml LB liquid medium, culture it on a shaker at 50°C for 24 hours, and obtain a seed solution.

[0139] c. Fermentation culture

[0140] The seed solution was inoculated into the fermentation medium at a volume ratio of 1:1000, and fermented at 50°C for 24 hours. The pressure of the fermenter was 0.04MPa, the ventilation rate was 0.2v / v.min, and the rotation speed was 200rpm. The fermented liquid was taken and centrifuged, the wet cells were collected, and freeze-dried at -50°C for 24 hours to obtain the freeze-dried powder of Bacillus subtilis H4.

[0141] The final concentration of the fermentation medium is c...

Embodiment 3

[0150] The character of embodiment 3 decomposing bacterial agents

[0151] (1) Protease activity

[0152] Prepare L-tyrosine standard solutions of 10, 20, 30, 40, 50, and 60 μg / ml, use double-distilled water without tyrosine as a blank control, and measure the absorbance at a wavelength of 275 nm with an ultraviolet spectrophotometer. Taking the absorbance as the ordinate and the concentration of tyrosine as the abscissa, a standard curve was drawn to obtain the equation y=0.0074x, and the absorption constant K was 135.1.

[0153] Take 10 g of the decomposed microbial agent prepared by the method in Example 2, add 100 ml of 0.1 mol / L phosphate buffer solution (pH=7.5), and shake on a shaker at 200 rpm for 30 min. Centrifuge at 3000rpm and take the decomposing agent supernatant.

[0154] Take 1.0ml of decomposing agent supernatant, keep it warm at 40°C for 2 minutes, add 1.0ml casein aqueous solution (10mg / ml), mix well and react at 40°C for 20min, add 2ml trichloroacetic aci...

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Abstract

The invention discloses a bacillus subtilis H4, a decomposed inoculant prepared therefrom and an application of the decomposed inoculant. The decomposed inoculants is composed of a freeze-dried powder of brevibacillus brevis H3,a freeze-dried powder of the bacillus subtilis H4, a freeze-dried powder of bacillus subtilis H8-3 and starch with a weight ratio being 1:1:1:97. The invention provides a new strain bacillus subtilis H4, the decomposed inoculant prepared from the bacillus subtilis H4 is high-temperature-resistant and can secrete protease and lipase for high-efficiently fermenting animal wastes. Compost prepared through the decomposed inoculant is less in environmental pollution, is economical and is easy to use. When a product after compost treatment is subjected to subsequent treatment, the product can be employed as an organic fertilizer for achieving an object of turning waste into resources.

Description

(1) Technical field [0001] The present invention relates to a decomposing bacterial agent, Bacillus subtilis strain and its application, in particular to a decomposing bacterial agent, the application of the decomposing bacterial agent in treating waste rich in protein and fat, and a new bacterial strain for preparing the decomposing bacterial agent-subtilis Bacillus H4. (2) Background technology [0002] Organic waste is often sent to landfills, incinerators or discarded as garbage. It is very necessary to develop methods for utilizing organic waste, not only to protect the environment, but also to turn waste into wealth. At present, many methods for utilizing organic waste have been developed, but most of them are for the utilization of waste with high cellulose and starch content, while there are fewer methods for the utilization of waste with high protein and fat content. [0003] Wastes rich in protein and fat (such as animal waste), if discarded directly, will genera...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F17/00C05F15/00C12R1/125C12R1/01
CPCC05F11/08C05F17/80C12N1/205C12R2001/01C12R2001/125Y02W30/40
Inventor 黄黎锋李海峰蓝袁洋施建军钟丹成沈如玥
Owner HANGZHOU FAMOXI BIOMEDICAL TECH
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