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Nitration brevibacillus nitrificans bacterial strain YJ1 and application thereof

A technology of nitrifying brevibacillus and brevibacillus, applied in the field of bioengineering, can solve the problem of less research fields and achieve good degradation characteristics

Active Publication Date: 2019-12-20
HUNAN HENGKAI ENVIRONMENT TECH INVESTMENT CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few research fields on the degrading strains of n-hexadecane at home and abroad. Only the patent CN 106242072A relates to the application of a non-degraded bacteria to degrade n-hexadecane. One of the technical problems in the field

Method used

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  • Nitration brevibacillus nitrificans bacterial strain YJ1 and application thereof
  • Nitration brevibacillus nitrificans bacterial strain YJ1 and application thereof
  • Nitration brevibacillus nitrificans bacterial strain YJ1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0082] Experimental example 1: Isolation, screening and purification of bacterial strains

[0083] Enrichment culture:

[0084] Take a 0-12cm soil sample from the oil depot of a relocated power parts factory and its surrounding oil plants. Add 10mL / L n-hexadecane as the only carbon source and energy source in the inorganic salt medium, add 10g of soil samples into a 250mL Erlenmeyer flask containing n-hexadecane 90mL inorganic salt liquid medium, place at 30 ℃, 180r / min constant temperature shaking incubator for 5 days, take 5mL of the mixed solution and fill it into fresh 95mL inorganic salt medium containing n-hexadecane. The above process was repeated several times to complete the enrichment of n-hexadecane degrading strains.

[0085] Screening of bacterial strains: Take a certain amount of the last enrichment culture solution and spread it on the inorganic salt agar medium containing n-hexadecane. After the plate is still for 30 minutes, place it upside down in a constan...

experiment example 2

[0092] Experimental example 2: Determination of growth curve and growth conditions of bacterial strain YJ1

[0093] Determination of growth curve: Pick a loop of the screened strains with an inoculation loop and dilute to OD with an appropriate amount of 0.85% normal saline 600 =0.1 to make a bacterial suspension, draw 1mL of the bacterial suspension and put it into 99mL BP liquid medium for shaking culture at 30°C and 180r / min, take a sample every 2 hours, due to the biomass and light absorption of the bacterial liquid in a fixed wavelength The value (OD) is proportional to the corresponding relationship, and the OD is measured with a spectrophotometer 600 value, in OD 600 Indicates the amount of growth of the strain, and 3 repetitions are made. The test measured the growth of n-hexadecane-degrading strain YJ1 in BP medium within 62 hours. The results showed that 0-10 hours was the lag phase of the strain, and 10-32 hours of cultivation was the logarithmic growth phase of t...

experiment example 3

[0098] Experimental Example 3: Analysis of Degradation Ability of Strain YJ1

[0099] According to the above test results, it was determined that the optimal growth conditions of the strain were temperature 30°C and pH 7.0, and the screened bacteria were inoculated in the corresponding BP liquid medium. 30°C, 180r / min constant temperature shaking culture for 24h. Plate count, the number of bacteria is 1.0×10 8 About CFU / mL, reserve.

[0100] Strain degradation test of n-hexadecane: Based on the liquid inorganic salt medium (pH=7) with the added concentration of 10mL / L n-hexadecane as the carbon source, the bacteria in the logarithmic growth phase were inoculated with 10% The amount was inserted into the culture medium, and the culture was shaken at a constant temperature at 30°C and 180 r / min. As a control, no strain was added, and the culture was shaken at a constant temperature for 5 days, 10 days, and 15 days.

[0101] The concentration of n-hexadecane was measured by ul...

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Abstract

The invention belongs to the biotechnology field, and discloses a hexadecane degradation bacterial strain YJ1 and an application thereof. A degradation bacterial strain using hexadecane as a carbon source is obtained through separating and screening. According to the shape, the physiological characteristic, the Gramstaining reaction, the 16S rDNA gene sequencing analysis and phylogenetic analysisof the bacterial strain, the bacterial strain is identified to be nitration Brevibacillus nitrificans, and is named as YJ1. The bacterial strain is preserved in the China Committee for Culture Collection of Microorganisms in the August, 2019, and the preservation No. is CGMCC NO.18418. The bacterial strain can use the hexadecane as the only carbon source, and the hexadecane can be rapidly degraded. After the initial concentration of the hexadecane is cultured at the environment of 10mg / L for 15 days, the degradation rate can achieve 66.7%. The appropriate growth condition of the bacterial strain is that the temperature is 30 DEG C, the pH is 7 and the inoculum concentration is 10%. The bacterial strain has good degradation characteristic for the hexadecane, and cannot cause secondary pollution to environment.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and specifically provides Brevibacillus nitrificans strain YJ1 and its application. Background technique [0002] With the rapid development of global industrialization, petroleum products have become the main source of energy in industrial production and daily life. At the same time, due to the rapid increase in oil consumption, long-term unscientific use, and leakage during transportation and mining, some hydrocarbons with strong toxicity, high residues, refractory, carcinogenic, and teratogenic substances have entered the soil and groundwater, causing environmental damage. cause pollution and seriously endanger human health. [0003] Most of the various hydrocarbons contained in petroleum, from the simplest compounds to complex solid residues with dozens of carbon atoms, can be metabolized and degraded by microorganisms, but the degree of difficulty and the speed of degradation are dif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10A62D3/02C12R1/01A62D101/20
CPCA62D3/02A62D2101/20B09C1/10C12N1/20C12N1/205C12R2001/01
Inventor 方雅瑜吴民熙郭照辉邢汉君罗容珺伍善东单世平冉启洋
Owner HUNAN HENGKAI ENVIRONMENT TECH INVESTMENT CO LTD
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