Grease degrading bacterium and application thereof

An oil and bacterial agent technology, which is applied to high-efficiency oil-degrading bacteria and the application field of dead livestock and poultry composting, can solve the problems of high oil content affecting the quality of chemical products, adverse effects of composting equipment, etc. Low, accelerated composting effect

Inactive Publication Date: 2017-11-24
HUNAN INST OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The currently developed composting bacteria agents are mainly aimed at the components in livestock manure and urban domestic waste, and the composting of dead livestock and poultry is different from other types of compost in various physical and chemical properties. The amount of organic fertilizer obtained by fermentation method is less than 20.3%, and the high oil content is the main factor affecting the quality of livestock and poultry carcass fertilizer products, and it also has an adverse effect on composting equipment

Method used

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  • Grease degrading bacterium and application thereof
  • Grease degrading bacterium and application thereof
  • Grease degrading bacterium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Enrichment, domestication and screening of oil-degrading bacteria

[0037] After diluting the soil collected from long-term oil-contaminated areas with sterile water 10 times, take 5mL of the diluted solution and add it to 45mL of the enrichment culture solution, at 30°C, 180r min -1 Shaking culture, every 4d is a cycle, inoculate 10mL of bacterial liquid into 90mL of new culture liquid to promote the growth of the strain, and the acclimatization lasts for 5 cycles. The initial refueling volume increases gradually with the cycle, respectively 0.8, 1.6, 2.4, 3.2, 4.0mL·L -1 .

[0038] Take 1 mL of the last domesticated bacterial solution and dilute it with sterile distilled water, and take 10–4 、10 –5 、10 –6 Three gradient bacterial solutions were applied to a neutral red oil plate, incubated at a constant temperature of 30°C for 3-5 days, and the growth of the colony and whether the surroundings of the colony turned red were observed. If it turns red, it ...

Embodiment 2

[0039] Example 2: Identification of Efficient Oil Degrading Bacteria KF-1

[0040] After morphological and microscopic observation of the bacterial strain KF-1 with the highest degradation rate, the genomic DNA of the bacteria was extracted with a DNA extraction kit, and 16S rDNA universal primer 27F was used for PCR amplification:

[0041] 5'-AGAGTTTGATCCTGGCTCAG-3', 1492R: 5'-GGTTACCTTGTTACGACTT-3'. PCR reaction conditions: pre-denaturation at 95°C for 3 min; denaturation at 94°C for 1 min, annealing at 56°C for 1 min, extension at 72°C for 2 min, and 30 cycles of reaction; final extension at 72°C for 10 min, and termination at 4°C. The PCR product was purified by a recovery kit and then submitted to Shanghai Bioengineering Co., Ltd. to determine its sequence, and then the sequence was submitted to the NCBI database for BLAST analysis.

[0042] After the strain KF-1 was cultured on the neutral red medium for 4 days, the surrounding area of ​​the colony became red obviously ...

Embodiment 3

[0043] Embodiment 3: The degradation ability of KF-1 to different oils and fats

[0044] Inoculate Bacillus subtilis KF-1 with 1% inoculum size in oil degradation medium containing 1% lard, peanut oil, soybean oil, and sesame oil, and cultivate it for 96 hours at 30°C and 180rpm, according to the method described in Example 1. The above method was used to calculate the degradation rate of strains to various oils. The degradation rate of KF-1 to the different oils and fats determined is as follows: Figure 4 shown. The results showed that KF-1 had the ability to degrade various oils, and its degradation effect on lard and peanut oil was significantly better than that of soybean oil and sesame oil.

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Abstract

The invention belongs to the technical field of microbiological degradation and provides a grease degrading bacterium and application thereof. According to the grease degrading bacterium disclosed by the invention, bacillus subtilis KF-1 is separated and screened from soil of an area polluted by grease, the strain is high temperature resistant and has stronger animal and vegetable grease degrading capacity and high lipase production activity, and lipase activity can reach 42.31U / ml after 72h shaking flask fermentation and cultivation. When a bactericide prepared from the bacillus subtilis KF-1 is applied to livestock dead body compost, compost maturity is accelerated, a degrease content in a compost product is obviously reduced, a seed germination index is improved, and quality of the compost product is improved.

Description

technical field [0001] The invention belongs to the technical field of microbial degradation, and relates to a high-efficiency oil-degrading bacterium and its application, especially the application in composting dead livestock and poultry. Background technique [0002] my country is a big country in animal husbandry, and its breeding scale ranks among the top in the world. Taking live pigs as an example, according to statistics, in 2015, there were 451 million live pigs in my country, and 708 million pigs were slaughtered. In the case of continuous expansion of breeding scale, the mortality of livestock and poultry caused by various diseases accounts for 5% to 10% of the total number of breeding every year, resulting in a huge number of infected animal carcasses. Diseased and dead livestock and poultry carry a large number of pathogens, which pose a serious threat to the healthy development of human beings and animal husbandry. How to harmlessly dispose of dead livestock ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C05F1/00C05F17/00C12R1/125
CPCC05F1/005C05F17/00C12R2001/125C12N1/205C05F11/08Y02A40/20Y02W30/40
Inventor 刘标刘惠知贺月林尹红梅许隽杜东霞王震许丽娟吴迎奔陈薇
Owner HUNAN INST OF MICROBIOLOGY
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