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Bacillus subtilis strain and application thereof

A technology of Bacillus subtilis and strains, applied in Bacillus subtilis strains and its application field, can solve the problems of narrow application area, water environment pollution, water resource shortage, etc., and achieve the effects of improving water quality, reducing COD, and simple cultivation conditions

Active Publication Date: 2016-10-12
浙江至美环境科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the lag of sewage treatment facilities and insufficient pollution control have caused global water resources to face two major problems: water pollution and water shortage
[0006] However, the above-mentioned strains are only aimed at organic pollutants in saponin wastewater, and their application is relatively narrow

Method used

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  • Bacillus subtilis strain and application thereof
  • Bacillus subtilis strain and application thereof
  • Bacillus subtilis strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The isolation and screening of embodiment 1 spore

[0027] 1. Collection of sludge

[0028] Collect 7 parts of sludge and water samples from the breeding pond, weigh 10g of sludge respectively (10mL of water sample is pipetted) into a conical flask filled with 90mL of sterilized normal saline, shake in a shaker at 150r / min for 0.5h, Take it out and put it in an 80°C water bath for 30 minutes, and dilute the sample solution by a certain number of times using the ten-fold dilution method, and set it aside.

[0029] Take 0.1 mL of diluted sample solution and spread it on solid LB medium, culture it at 30°C for 48 hours, select the colonies with good growth, similar colony shape and the largest number, separate and purify until the single bacteria with the same colony shape, and obtain 12 strains of spores.

[0030] 2. Screening of Enzyme-producing Bacillus

[0031] (1) Cellulase detection:

[0032] Carboxymethylcellulose sodium plate medium (g / L): CMC-Na 15.0, NaCl 5.0,...

Embodiment 2

[0045] The identification of embodiment 2 spore CM-A12

[0046] (1) Morphological characteristics

[0047] After the strain CM-A12 was cultured on the solid LB growth medium for 24 hours, the colony was in the shape of transparent water droplets, with a raised surface, moist and shiny; after 48 hours of culture, the surface of the colony was dry and off-white. Under the microscope, it was observed that the strain cells were Brevibacteria, such as figure 1 shown.

[0048](2) Physiological and biochemical identification (as shown in Table 2)

[0049] Table 2

[0050]

[0051]

[0052] Note: "+": the reaction is positive; "-" the reaction is negative.

[0053] (3) 16S rDNA determination

[0054] The 16S rDNA gene sequence of the strain could not be determined by observing the morphological characteristics of the strain, and the identification at the molecular level was carried out.

[0055] PCR amplification of 16S rDNA of strain a

[0056] ①PCR amplification system:...

Embodiment 3

[0067] Embodiment 3 Bacillus subtilis (Bacillus subtilis) CM-A12 growth condition optimization

[0068] Strain culture medium: peptone 10g / L, beef extract 5g / L, NaCl 5g / L, pH 7-7.2.

[0069] (1) Optimum growth pH of the inventive bacterial strain

[0070] The initial pH of the culture solution was adjusted to 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, 11.0 with 1.0 mol / L HCl and NaOH, respectively, and the CM-A12 was inserted into 1% (v / v) inoculum, Cultivate at 30°C and 150r / min. After culturing for 20h and 40h respectively, measure the absorbance at 600nm with an ultraviolet-visible spectrophotometer, and use the culture solution without inoculation as a blank control.

[0071] Depend on image 3 It is known that CM-A12 grows well when the initial pH is between 4 and 9, and the pH range is wide; when the initial pH value is greater than 9.0, the strain CM-A12 can hardly grow.

[0072] From an economic point of view, when CM-A12 is mass-produced and fermented, the medium should b...

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Abstract

Belonging to the field of environmental microorganism application, the invention discloses a Bacillus subtilis strain and application thereof. The strain is named as Bacillus subtilis CM-A12, is currently preserved in China Center For Type Culture Collection (called CCTCC for short) with a preservation number of CCTCC NO:M 2015688, and the preservation date is November 23, 2015. The Bacillus subtilis strain provided by the invention has the advantages of simple cultivation condition, high temperature resistance, acid and alkali resistance, and high salinity resistance, can secrete protease, amylase and cellulase, and can effectively improve water quality and reduce COD and ammonia nitrogen in water.

Description

technical field [0001] The invention relates to the application field of environmental microorganisms, in particular to a strain of bacillus subtilis and its application. Background technique [0002] With the development of social economy, the increase of population and the continuous improvement of people's living standards, the demand and consumption of water resources are constantly increasing, and the corresponding discharge of urban sewage and industrial wastewater is also constantly increasing. However, the lagging of sewage treatment facilities and insufficient pollution control have caused global water resources to face two major problems: water environmental pollution and water resource shortage. In some countries and regions, especially developing countries including China, it is becoming more and more serious and has a tendency to intensify. In order to ensure the sustainable development of human society and economy, water environment protection and governance h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N11/00C02F3/34C12R1/125
CPCC02F3/34C12N1/20C12N11/00C12N1/205C12R2001/125
Inventor 赵鸭美朱强李美娣王晛唐青青
Owner 浙江至美环境科技有限公司
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