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Method for efficiently and directionally enriching and separating hydrogen producing bacteria

A directional enrichment and high-efficiency technology, applied in the field of microorganisms, to achieve the effects of low energy consumption, high substrate conversion efficiency, and simple synthesis method

Active Publication Date: 2019-08-09
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the method of directional, fast and convenient enrichment and separation of Clostridium

Method used

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  • Method for efficiently and directionally enriching and separating hydrogen producing bacteria
  • Method for efficiently and directionally enriching and separating hydrogen producing bacteria
  • Method for efficiently and directionally enriching and separating hydrogen producing bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Method for efficient directional enrichment and isolation of hydrogen-producing bacteria:

[0047] 1) Anaerobic sludge heat treatment and incubation under oligotrophic induction conditions:

[0048] Wastewater activated sludge was obtained from a full-scale upflow anaerobic sludge bed (UASB) reactor for wastewater treatment in a brewery and stored in a 4 °C refrigerator until use. Among them, the pH of activated sludge: 5-7, solid concentration (MLSS) is 50-55mg / L, soluble total organic carbon (STOC) is 2000mg / L;

[0049] (1) Mix 50g of activated sludge with 500mL of inorganic salt basal medium evenly, and add 2g of straw powder as a substrate to obtain a slurry suspension;

[0050] (2) Pack the mixed mud suspension and divide it into an experimental group and a control group. The experimental group adds amorphous ferrihydrite with a final concentration of 100mg / L, and the control group adds an equal volume of PBS solution without adding ferrihydrite. mine;

[0051] ...

Embodiment 2

[0062] Based on the high-throughput sequencing analysis of the 16S rRNA gene, the community structure of the above-mentioned high-hydrogen-producing bacteria was obtained:

[0063] In the enrichment products SF1-SF3 of the experimental group and the enrichment products S1-S3 of the control group at the phylum level, Firmicutes are dominant phyla such as figure 2 (a), its abundance is as high as 85%, such as figure 2 (b) shows the results; while the dynamic analysis of the community structure at the genus level shows that the abundance of the main hydrogen-producing bacteria genus Clostridium in the experimental group SF1-SF3 increases from 1% to 27.69% with the enrichment and passage process gradually increased to 77.4%, that is to say, the hydrogen-producing Clostridium gradually changed from a rare species to an absolute dominant genus during the enrichment induction process, and the results were as follows: figure 2 as shown in b. Given that Clostridium is the main gen...

Embodiment 3

[0065] Further purification and separation of the above-mentioned hydrogen-producing colonies:

[0066] After the suspension of the hydrogen-producing enriched bacterial population is serially diluted (in order to obtain a single colony of the hydrogen-producing strain), it is evenly spread on the surface of the MSG solid medium supplemented with 2% agarose, and incubated in the dark at 30-37°C until Monoclonal colonies appear on the surface of the culture medium, which is the high-efficiency hydrogen-producing strain Clostridium pasteurianum YC-1.

[0067]The obtained strains were subjected to colony PCR, and a single colony was selected and inserted into 20 μL sterile water, and 2 μL was used for PCR. PCR reaction system: 2 μL buffer buffer; 2 μL dNTP (2.5mol / L); 0.5 μL Taq enzyme (5U / μL); 0.5 μL Ba907R (20 μmol / L, 5′-AGA GTTTGATCCTGG CTCAG-3′) reverse primer; 0.5 μL Ba27F (20 μmol / L, 5′-CCGTCAATTCCTTTRAGTTT-3′) forward primer; 2 μL single colony suspension template; 2.5 μL...

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Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to a method for efficiently and directionally enriching and separating hydrogen producing bacteria and application of the hydrogen producing bacteria obtained through separation in hydrogen production and acetic acid and butyric acid production through fermentation. According to the method, after heat treatment is performed on activated sludge, an inducer amorphous ferrihydrite is added, and incubation is performed under an oligotrophic condition; an incubated culture is transferred into an MSG-rich culture medium containing the amorphous ferrihydrite for cultivation, passage culture continues to be carried out on a cultured product, and a product is a hydrogen producing enriched flora. The method forefficiently and directionally enriching and separating the hydrogen producing bacteria can achieve the efficient and convenient enrichment and separation of the hydrogen producing bacteria or strains,and has important guiding significance and broad application prospects in the industrial production process of microbial fermentation hydrogen production.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a method for highly efficient directional enrichment and separation of hydrogen-producing bacteria and the application of the hydrogen-producing bacteria to hydrogen production, acetic acid production and butyric acid production. Background technique [0002] Hydrogen is an important clean energy with a high heat of combustion (122kJ / g). Due to the growing problems of non-renewable fossil fuel consumption and the climate change and air pollution caused by its use, hydrogen shows great application prospects. At present, countries around the world have taken the development of hydrogen energy as one of the important strategies for the country's future energy. The 2018 "World and China Energy Outlook 2050" pointed out that my country's energy development has entered a period of transition from old to new kinetic energy. The "Shandong Province New Energy Industry Devel...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02C12P3/00C12P7/54C12P7/52C12R1/145
CPCC12N1/20C12N1/02C12P3/00C12P7/54C12P7/52C12R2001/145C12N1/205Y02E50/10
Inventor 刘芳华张月超肖雷雷
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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