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Pseudomonas mendocina NX-1 and application thereof to degradation of n-hexane

A technology of Pseudomonas syringae and Monas, which is applied in the field of high-efficiency new-type n-hexane degrading bacteria, can solve the problems of few reports, screening, and less n-hexane, and achieve no secondary pollution, stable operation, and low operating costs low effect

Active Publication Date: 2015-11-25
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few studies on the biodegradation of n-hexane in China at present, and there are few reports to screen out highly efficient degradation strains using n-hexane as the sole carbon source

Method used

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  • Pseudomonas mendocina NX-1 and application thereof to degradation of n-hexane
  • Pseudomonas mendocina NX-1 and application thereof to degradation of n-hexane
  • Pseudomonas mendocina NX-1 and application thereof to degradation of n-hexane

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Experimental program
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Effect test

Embodiment 1

[0028] Embodiment 1: Isolation and identification of Pseudomonas mendoza NX-1

[0029] (1) Sample collection and domestication

[0030] Preliminary screening: Collect activated sludge from the sewage treatment tank of a pharmaceutical and chemical factory in Zhejiang on-site. After standing for 2 hours, the supernatant and suspended impurities are removed, leaving fine particles of sludge. The lower sludge after standing is mixed with Inorganic salt medium was mixed 1:1 (v / v) into the sludge acclimatization tank, with a total volume of 5 L, with n-hexane as the substrate as the sole carbon source and energy source, (30±1°C) constant temperature acclimatization culture, The sludge is domesticated, and the domestication tank is stably aerated during the domestication experiment, and the pH value of the domestication tank is controlled to maintain at 7.0. When the substrate concentration drops below 50mg / L every day, add the substrate once to make the substrate concentration after ...

Embodiment 2

[0039] The seed liquid preparation of embodiment 2 Pseudomonas mendoza NX-1 bacterial strain

[0040] 1) Slant culture: inoculate Pseudomonas mendoza NX-1 on R 2 A solid slant culture medium, cultured at 30-32°C for 36-48 hours to obtain slant bacteria, the R 2 The final concentration of A solid slant medium is composed of: yeast powder 0.50g / L, casein 0.50g / L, soluble starch 0.50g / L, MgCl 2 ·7H 2 O0.05g / L, tryptone 0.50g / L, glucose 0.50g / L, sodium pyruvate 0.30g / L, KH 2 PO 4 0.45g / L, agar 15~18g / L, pH 7.2, solvent is water.

[0041] 2) Seed culture: use an inoculation loop to pick the slant bacteria obtained in step (1) and inoculate it into the seed medium for cultivation, add n-hexane with a final concentration of 100-400 mg / L, cultivate at 30-32°C for 36-38 hours, and obtain OD 600 =0.1~0.3 bacterial liquid, the seed culture medium is an inorganic salt culture medium.

Embodiment 3

[0042] Embodiment 3: the performance of Pseudomonas mendoza NX-1 (CCTCCNO: M2015114) degradation n-hexane

[0043] With normal hexane as the sole carbon source of Pseudomonas mendoza NX-1, get the bacterium liquid prepared by the method of embodiment 2, inoculate to fresh 50mL inorganic salt culture medium containing 105mg / L normal hexane, make initial thalline Concentration (in OD 600 Meter) is 0.02, pH=7. Put it into a shaker with a temperature of 30°C and a rotation speed of 160r / min for cultivation, take samples every 3h to 4h to measure the degradation rate of n-hexane, and use a 5mL syringe to draw out a part of the bacterial liquid to measure the OD value of the bacterial body. The results are shown in image 3 . During the experiment, two parallel samples and a blank control group not inoculated with strains were designed. With the prolongation of time, the bacterial cell concentration gradually increased, and at 30 hours, the bacterial cell concentration reached a ...

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Abstract

The invention provides a novel efficient n-hexane degrading bacterium-pseudomonas mendocina NX-1 and an application thereof to microbial degradation of n-hexane. Pseudomonas mendocina NX-1 is collected in the China Center for Type Culture Collection (CCTCC) in Wuhan University in China, 430072, with collection date of March 15, 2015 and collection number of CCTCC NO:M2015114. The n-hexane degrading bacterium provided by the invention is an aerobic non-fermentation Gram-negative bacterium and can utilize n-hexane as an only carbon source for reproduction and completely mineralize the substrate into CO2 and H2O. Under the condition of pure culture, the bacterium can be used for degrading n-hexane in the pH value range of 4.0-10.0. The strain has stronger adaptive capacity to environment and lays the foundation for engineering application to purifying n-hexane containing wastewater and waste gases by a biological method.

Description

(1) Technical field [0001] The invention relates to a strain of high-efficiency novel n-hexane degrading bacteria—Pseudomonas mendocina NX-1 and its application in microbial degradation of n-hexane. (2) Background technology [0002] n-hexane (n-hexane, C 6 h 6 ) is a colorless liquid with low toxicity and a faint special smell. It is a chemical solvent, mainly used as a solvent for the polymerization of olefins such as propylene, as an extractant for edible vegetable oil, as a solvent for rubber and paint, and as a thinner for pigments. It has certain toxicity and will enter the human body through the respiratory tract, skin, etc. Long-term exposure can lead to chronic poisoning symptoms such as headache, dizziness, fatigue, and numbness of limbs, and severe cases can lead to fainting, loss of consciousness, and even death. [0003] Due to the wide application of n-hexane and long-term inhalation of n-hexane can lead to chronic poisoning, the development of economical an...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/38C02F101/32
Inventor 陈东之陈建孟江宁馨叶杰旭
Owner ZHEJIANG UNIV OF TECH
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