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Low-temperature-resistant methanogen enrichment culture method and application thereof

A technology of enrichment culture and methanogenic bacteria, which is applied in the direction of using microorganisms, waste fuels, fermentation, etc., can solve the problems of high time cost and economic cost of bacterial agents, decreased biogas fermentation effect, and long time required, etc., to achieve Low temperature application Wide temperature range, few steps and short time effect

Inactive Publication Date: 2016-05-25
HARBIN INST OF TECH
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  • Summary
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Problems solved by technology

[0003] To solve the problem of the decline of biogas fermentation effect at low temperature, the development of low-temperature methanogenic bacteria is a fundamental solution to the problem
The usual method of obtaining methanogenic bacteria is the traditional method: first obtain pure bacteria through enrichment, separation and purification, and screening, and then carry out strain compounding, or directly use low temperature to domesticate methanogenic bacteria, and continue to subculture, the steps It is cumbersome and takes a long time, which leads to high time cost and economic cost for bacterial agents from acquisition, production to application

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  • Low-temperature-resistant methanogen enrichment culture method and application thereof
  • Low-temperature-resistant methanogen enrichment culture method and application thereof
  • Low-temperature-resistant methanogen enrichment culture method and application thereof

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specific Embodiment approach 1

[0018] Specific embodiment 1: The method for enrichment and cultivation of low-temperature-resistant methanogens of this embodiment is carried out according to the following steps:

[0019] Take the activated sludge and break it, put it into the enrichment medium, and conduct the enrichment culture under the condition of the enrichment culture temperature of 15~25℃; when it is detected that the enrichment culture system generates bubbles, pass the control The temperature system gradually lowers the temperature until no bubbles are detected, and the temperature is stopped to complete the enrichment culture of low-temperature resistant methanogens.

[0020] The present invention finds that the bacteria before and after enrichment are mainly Bacteroidetes, Firmicutes, Proteobacteria, Spirochaete, and Chloroflexus by analyzing the structure of the bacteria before and after enrichment. (Chloroflexi), Synergistetes, Acidobacteria, Lentisphaerae and Fibrobacteres. After enrichment of met...

specific Embodiment approach 2

[0021] Specific embodiment two: this embodiment is different from specific embodiment one in that the activated sludge is a natural habitat wetland sludge, activated sludge from a sewage treatment plant or livestock and poultry manure, and a mixture of biogas slurry and biogas residue. Others are the same as the first embodiment.

specific Embodiment approach 3

[0022] Specific embodiment three: This embodiment is different from the specific embodiment one in that the components of the enrichment medium contain 0.5 g of K per 1L 2 HPO 4 ·3H 2 O, 0.4g / L KH 2 PO 4 , 1g of NH 4 Cl, 0.2g of MgCl 2 ·6H 2 O, 2g of yeast powder, 2g of peptone, 5g of sodium formate, 8.3g of sodium acetate, 3mL of methanol, 0.5g of cysteine, 1ml of vitamin solution and 1ml of trace element solution; pH value is 6.8-7.2 ;

[0023] Wherein, the trace element liquid contains 1.5g of FeCl per 1L 2 , 70mg of ZnCl 2 , 6mg boric acid, 0.1g MnCl 2 ·4H 2 O, 2mg CuCl 2 ·2H 2 O, 0.19g of CoCl 2 ·6H 2 O, 24mg NiCl 2 ·6H 2 O, 36mg of Na 2 MO 4 ·H 2 O, 15mg of Na 2 WO 4 ·2H 2 O and 15mg of Na 2 SeO 4 ·5H 2 O composition;

[0024] The vitamin solution contains 50.0 mg of lipoic acid, 20.0 mg of biotin, 0.35 g of niacin, 5.0 mg of thiamine hydrochloride, 50.0 mg of para-aminobenzoic acid, 20.0 mg of folic acid, and 50.0 mg of pantothenic acid per 1L. Calcium, 1.0mg of vitamin B 12...

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Abstract

The invention relates to a low-temperature-resistant methanogen enrichment culture method and an application thereof. The low-temperature-resistant methanogen enrichment culture method comprises the following steps: firstly, taking activated sludge, breaking into pieces, inoculating into an enrichment culture medium, and determining enrichment culture temperature at the temperature of 15-25 DEG C according to sources of inocula; and secondly, when bubbles are detected in a culture system, gradually lowering temperature by virtue of a temperature control system, and stopping temperature lowering until no bubble is detected, so that low-temperature-resistant methanogen enrichment culture is completed. An application mode comprises that enrichment culture products are added into an accelerator prepared from ferric chloride, cobalt chloride, nickel chloride and manganese chloride and the obtained mixture is directly applied to low temperature methane fermentation. The low-temperature-resistant methanogen enrichment culture method can be used for solving the problems that preparation steps of an existing low temperature methanogen agent are tedious, low temperature domestication time is long and status of dominant bacteria is difficultly maintained for a long time at a low temperature. The low-temperature-resistant methanogen enrichment culture method is simple and easy to operate, has the advantages of short enrichment time, stable performance, good and stable running effect, available flora source and the like and is convenient for popularization and application.

Description

Technical field [0001] The invention belongs to the technical field of clean energy production by microbial fermentation, and specifically relates to a method for enrichment and cultivation of low-temperature-resistant methanogenic bacteria and low-temperature application thereof. Background technique [0002] The exhaustion of fossil fuel reserves and continuous environmental pollution have become major problems facing the world today. With population growth and economic development, global energy demand will continue to rise in the future. The dual crisis of energy and environment has prompted countries around the world to actively seek renewable and environmentally friendly alternatives to fossil fuels-biomass energy. As a kind of renewable clean biomass energy, biogas has broad development prospects, and has been widely concerned and applied worldwide. Biogas fermentation is a complex process. The fermentation process is affected by conditions such as temperature, moisture,...

Claims

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Application Information

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IPC IPC(8): C12N1/36C12P5/02
CPCC12N1/36C12P5/023Y02E50/30
Inventor 曹广丽张军政杜洋宋金柱丛华
Owner HARBIN INST OF TECH
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