Denitrifying bacterium and fermenting production method thereof

A technology of denitrifying bacteria and production methods, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of high energy consumption, cumbersome industrial fermentation procedures, low optimal temperature, etc., and achieve simple fermentation process , high live bacteria content and spore rate, and low cost

Active Publication Date: 2015-01-28
CHINA PETROLEUM & CHEM CORP +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the problems in the prior art that the optimal temperature for denitrifying bacteria to inhibit SRB is low, the industrial fermentation procedure of bacterial liquid is cumbersome and the energy consumption is high, and a denitrifying bacteria is provided to increase the inhibition temperature of denitrifying bacteria inhibiting SRB, And it can simplify the industrial fermentation process of denitrifying bacteria

Method used

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  • Denitrifying bacterium and fermenting production method thereof
  • Denitrifying bacterium and fermenting production method thereof
  • Denitrifying bacterium and fermenting production method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The screening steps of JSHD-1 denitrifying bacteria of the present invention are as follows:

[0019] (1) Sample collection

[0020] The produced fluids of 3 oil wells were collected in Jiangsu Oilfield, and the sample names were: Fu 129, Fu 137, and Fu 146.

[0021] (2) Culture medium configuration

[0022] The enrichment medium is composed of A solution and B solution, in which:

[0023] Solution A: 2.0 g inorganic salt K1, 1.0 g asparagine, 500 ml distilled water, pH 7.0~7.2;

[0024] Solution B: 8.5 g sodium citrate, 2 g sodium acetate, 2 g glucose, KH 2 PO 4 0.5 g, K 2 HPO 4 0.5 g, MgSO 4 ·7H 2 O 1.0 g, FeCl 3 ·6H 2 O 0.05 g, CaCl 2 0.1 g, yeast extract 0.5 g, distilled water 500 ml. The pH value is 7.0~7.2.

[0025] NRB liquid medium: liquid A and liquid B are sterilized separately at 121 ℃ for 20 min, and mixed in equal proportions after sterilization.

[0026] NRB solid medium: add agar with a mass ratio of 1.5% to 2% to liquid A and liquid B, a...

Embodiment 2

[0065] (1) Inoculate CGMCC NO 7270 strains on 3 eggplant bottles filled with solid medium. The solid medium formula and the mass percentage of each component are: sodium nitrate 0.2%, sodium acetate 0.1%, potassium dihydrogen phosphate 0.1%, glucose 0.1%, magnesium sulfate 0.02%, aspartic acid 0.05%, ferric chloride 0.01%, yeast extract 0.012%, agar 2.0%, the balance is water, pH value 7.0. The inoculated solid medium was cultured statically in a 60°C incubator for 3 days to form denitrifying bacterial spores.

[0066] (2) Wash the surface of the solid medium with sterile water to obtain 250mL of denitrifying bacterial spore suspension, and the content of spores counted by microscope hemocytometer is 2.7×10 10 cfu / mL.

[0067] (3) The above-mentioned spore suspension was pretreated in a 70°C water bath for 10 minutes, and then inoculated into the sterile medium of a 10,000L fermenter. The medium formula and the mass percentages of each component were: sodium nitrate 0.2%, so...

Embodiment 3

[0069] (1) Inoculate CGMCC NO 7270 strains on 3 eggplant bottles filled with solid medium. The solid medium formula and the mass percentage of each component are: sodium nitrate 0.2%, sodium acetate 0.1%, potassium dihydrogen phosphate 0.1%, glucose 0.1%, magnesium sulfate 0.02%, aspartic acid 0.05%, ferric chloride 0.01%, yeast extract 0.012%, agar 2.0%, the balance is water, pH value 7.0. The inoculated solid medium was cultured statically in a constant temperature incubator at 50°C for 4 days to form denitrifying bacterial spores.

[0070] (2) Wash the surface of the solid medium with sterile water to obtain 250 mL of denitrifying bacterial spore suspension, and the content of spores counted by microscope hemocytometer is 3.5×10 10 cfu / mL.

[0071] (3) The above-mentioned spore suspension was pretreated in a 70°C water bath for 10 minutes, and then inoculated into the sterile medium of a 10,000L fermenter. The medium formula and the mass percentages of each component were...

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Abstract

The invention relates to a denitrifying bacterium which can be used for effectively utilizing an inorganic nitrogen source and inhibiting growth and reproduction of sulfate reducing bacteria (SRB) and a fermenting production method of the denitrifying bacterium, belonging to the technical field of microbial fermentation engineering. The high-temperature denitrifying bacterium provided by the invention is preserved in CGMCC, the Latin name of the high-temperature denitrifying bacterium is Geobacillus pallidas, the preservation number is CGMCC NO 7270 and the preservation date is 5th, March, 2013. The high-temperature denitrifying bacterium provided by the invention can be used for effectively utilizing the nitrogen source and effectively inhibiting growth and reproduction of SRB under a high temperature condition of 45-90 DEG C. The denitrifying bacterium is simple in fermentation process and can be successfully fermented, and the culture medium is low in cost and short in culture period. The produced denitrifying bacterium is high in viable organism content and high in germ rate. The content of unit germ cells reaches 2*10<8>cfu/mL and the content of germs reaches over 90%.

Description

technical field [0001] The invention relates to the technical field of microbial fermentation engineering, in particular to a denitrifying bacterium capable of effectively utilizing inorganic nitrogen sources and inhibiting the growth and reproduction of sulfate-reducing bacteria (SRB) and a fermentation production method of the denitrifying bacterium. Background technique [0002] Waterflooding development of oil reservoirs leads to biodegradation of crude oil and its intermediate products, which stimulate the growth of sulfate-reducing bacteria (SRB) in oil reservoirs, oil pipelines and oil production units. Sulfate-reducing bacteria produce a large amount of hydrogen sulfide, which causes acidification of oil reservoirs and corrosion of pipelines, which has brought serious losses to the global petroleum industry. Recent data from scientific research institutions in the United States and West Germany show that: SRB corrosion only causes losses of 700 million US dollars per...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/01
CPCC12N1/20C12N1/205C12R2001/01
Inventor 吴伟林杨海滨姚峰林晶晶孟章进杨帆
Owner CHINA PETROLEUM & CHEM CORP
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