Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for separating nitrogen-fixing bacteria from petroleum pollutants

A technology for oil pollution and nitrogen-fixing bacteria, applied in microorganism-based methods, biochemical equipment and methods, separation of microorganisms, etc. performance, improved accuracy and separation efficiency

Inactive Publication Date: 2018-10-09
CHINA AGRI UNIV
View PDF3 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many types of autogenous nitrogen-fixing bacteria, including aerobic, anaerobic, micro-aerobic, facultative anaerobic, and photosynthetic bacteria. Because nitrogenase is highly sensitive to oxygen, many nitrogen-fixing bacteria can only be produced under anaerobic or micro-aerobic conditions. Nitrogen fixation, easily missed in conventional separation methods
Therefore, the conventional nitrogen-free medium surface coating method can only obtain some aerobic nitrogen-fixing bacteria with an oxygen-defense protection mechanism, and other types of nitrogen-fixing bacteria cannot fix nitrogen under aerobic conditions due to the different oxygen requirements of nitrogenase. , thus losing the ability to grow in nitrogen-free media and thus being missed in the screen

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for separating nitrogen-fixing bacteria from petroleum pollutants
  • Method for separating nitrogen-fixing bacteria from petroleum pollutants
  • Method for separating nitrogen-fixing bacteria from petroleum pollutants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The growth situation of embodiment 1 nitrogen-fixing bacteria adding different concentrations of glucose in nitrogen-free medium

[0036] (1) Bacterial culture: Select the bacterial strain Klebsiella sp PJ35 with confirmed nitrogen-fixing ability as the experimental material, use LB medium to cultivate the bacterial strain to the logarithmic phase, and adjust the initial density to OD 600 = 0.2;

[0037] (2) Detection of growth ability: the cultured nitrogen-fixing bacteria were inoculated into nitrogen-free medium containing different glucose concentrations, and the glucose concentrations were respectively: 10g / L, 30g / L, 50g / L, and 80g / L. Measure the OD of the bacteria every two hours 600 value. see results figure 1 .

[0038] It shows that under the concentration of 30g / L and 50g / L glucose, the growth ability of nitrogen-fixing bacteria in the nitrogen-free medium is optimal, and after the concentration of glucose exceeds 80g / L, its growth will be inhibited, while...

Embodiment 2

[0039] Example 2 Isolation of Nitrogen-fixing Bacteria from Soil Contaminated by Explosion of Huangwei Oil Pipeline Explosion in Huangdao District, Qingdao City

[0040] (1) Sample collection: Collect the crude oil contaminated soil from the explosion of Qingdao Huangwei Oil Pipeline for screening of nitrogen-fixing bacteria

[0041] (2) Isolation of nitrogen-fixing bacteria: Weigh 2.5g soil sample and add 10ml sterile water to suspend, vortex shaker for 10min to promote dispersion, stand still for 2-5 minutes, absorb the suspension, dilute to 10 -3 , Melt the nitrogen-free solid medium and keep it warm at 55°C. Take 1ml of each dilution and add it to 25ml nitrogen-free solid medium. After mixing thoroughly, prepare a plate immediately, and repeat each dilution 3 times. After culturing at 30°C for 2-4 days, there will be colonies growing on the surface or inside of the medium, that is, the nitrogen-fixing bacteria in the oil-contaminated soil were isolated, and a single colony...

Embodiment 3

[0049] Example 3 Isolation of nitrogen-fixing bacteria in Xinmin high pouring oil contaminated soil

[0050] (1) Sample collection: oil-contaminated soil was collected from oil fields.

[0051] (2) Isolation of nitrogen-fixing bacteria: Weigh 2.5g soil sample and add 10ml sterile water to suspend, vortex shaker for 10min to promote dispersion, stand still for 2-5 minutes, absorb the suspension, dilute to 10 -3 , Melt the nitrogen-free solid medium and keep it warm at 55°C. Take 1ml of each dilution and add it to 25ml nitrogen-free solid medium. After mixing thoroughly, prepare a plate immediately, and repeat each dilution 3 times. After culturing at 30°C for 2-4 days, there will be colonies growing on the surface or inside of the medium, that is, the nitrogen-fixing bacteria in the oil-contaminated soil were isolated, and a single colony was picked and purified on the GMCY medium, and stored at -80°C.

[0052] (3) Analysis of phylogenetic status of nitrogen-fixing bacteria: a...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a method for separating nitrogen-fixing bacteria from petroleum pollutants, and belongs to the technical field of microorganism separation culture. The method comprises the following steps: (1) collecting the petroleum pollutants, adding water for suspension, and carrying out gradient dilution on suspension liquid; (2) preparing a nitrogen-free solid medium, separately adding suspension liquids with different dilutions when the medium is in a melted state, blending and then standing for culturing, and if bacterial colonies grow on the surface or in the medium, separatingthe bacterial colonies to obtain the nitrogen-fixing bacteria in the petroleum pollutants. The problems that bacteria capable of fixing nitrogen under an aerobic condition only can be obtained by a conventional method, and bacteria capable of fixing nitrogen under an microaerobic or anaerobic condition are easily missing during screening can be solved, diversified demands of the nitrogen-fixing bacteria to oxygen are met, therefore, the accuracy and efficiency of separation of the nitrogen-fixing bacteria from the petroleum pollutants are improved, and the application prospect is wide.

Description

technical field [0001] The invention relates to the technical field of microorganism isolation and cultivation, in particular to a method for isolating nitrogen-fixing bacteria from oil-polluted soil or oil-polluted water. Background technique [0002] Biological nitrogen fixation refers to the process of nitrogen-fixing microorganisms converting nitrogen in the atmosphere into a nitrogen source that can be used by organisms. It can be divided into symbiotic nitrogen fixation, joint nitrogen fixation and autogenous nitrogen fixation. Symbiotic nitrogen fixation generally means that microorganisms live closely with other organisms and form a special symbiotic structure for nitrogen fixation; combined nitrogen fixation means that some autogenous nitrogen-fixing bacteria can colonize the rhizosphere or body of plants, and there is a mutually beneficial relationship with the host. And have a certain specificity; autogenous nitrogen-fixing bacteria usually live independently in t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N1/02C12N1/20C12R1/065
CPCC12N1/02C12N1/20
Inventor 王磊陈琳张心昱郑国砥徐志凌
Owner CHINA AGRI UNIV
Features
  • Generate Ideas
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com