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Minimal gingivalis T7-7 degrading hydrocarbon under low temperature and uses thereof

A micromonas, strain technology, applied in the field of microbial biotechnology and environmental biology, can solve problems such as marine oil pollution and serious problems, and achieve the effect of effective degradation

Inactive Publication Date: 2008-07-23
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the increasingly serious problem of marine oil pollution, provide a kind of micromonas T7-7 (Pusillimonas sp.T7-7) and the application of this bacterium in the bioremediation of marine oil pollution

Method used

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  • Minimal gingivalis T7-7 degrading hydrocarbon under low temperature and uses thereof
  • Minimal gingivalis T7-7 degrading hydrocarbon under low temperature and uses thereof
  • Minimal gingivalis T7-7 degrading hydrocarbon under low temperature and uses thereof

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Experimental program
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Effect test

Embodiment 1

[0029] Screening and breeding of the Pusillimonas sp. T7-7 strain provided by the present invention.

[0030] Mud samples were collected from the oil-polluted seabed area of ​​the Bohai Sea. The latitude and longitude coordinates are 118°26′36″ east longitude and 38°50′30″ north latitude. Get 5g mud sample in 100mL liquid paraffin as the inorganic salt culture medium of sole carbon source (substrate composition: KH 2 PO 4 3.48, Na 2 HPO 4 12H 2 O 1.5, NH 4 SO 4 2. MgSO 4 ·7H 2 O 0.70, yeast extract 0.05; liquid paraffin content 2%; distilled water, 1000ml; pH7.2; sterilized at 121°C for 30min), placed on a 200r / min low-temperature shaker at 15°C for enrichment culture for 7 days, and obtained 7 strains by marking The mixed flora of the bacteria was named as T7-1~T7-7; 5mL of the enrichment solution was taken and transferred to a fresh 100mL liquid paraffin medium for fermentation and culture, and the culture conditions were the same as above; after three cycles of e...

Embodiment 2

[0044] The morphological characteristics and physiological and biochemical characteristics of the Pusillimonas sp. T7-7 bacterial strain provided by the present invention.

[0045] According to the experimental method of "Bergey's Manual of Systematic Bacteriology" (Vol. VIII), the Gram staining, cell size and shape, presence or absence of flagella and spores, growth temperature, growth pH range, and NaCl tolerance were detected. Catalase, oxidase, M.R. experiment, V-P experiment, indole production, nitrate reduction, nitrite reduction, denitrification, starch hydrolysis, ammonia production experiment, urease, hydrogen sulfide production, gelatin hydrolysis, glucose fermentation, litmus milk Experiment, decomposition of aescin experiment.

[0046] Gram staining is negative, the bacteria are short rod-shaped, single-ended polar flagella, able to move, facultatively aerobic, and the cell size is 0.2-0.5μm (width)×0.5-1.0μm (length). The growth temperature is 4-37°C, the growth ...

Embodiment 3

[0048] Analysis of cell wall fatty acids of the Pusillimonas sp. T7-7 strain provided by the present invention.

[0049] Take an appropriate amount of bacterial culture, put it in an 8ml screw-top glass tube, add 1ml solution I (45g sodium hydroxide dissolved in 150ml methanol and 150ml distilled water), tighten the screw cap, put it in a boiling water bath for 5min, take it out and shake it for 5-10 seconds, and tighten it again Screw the cap on, and continue the boiling water bath for 25 minutes; after the sample tube is cooled, add 2ml of solution II (190ml of concentrated hydrochloric acid, 275ml of methanol dissolved in 135ml of distilled water), cover tightly and shake, then precisely control the water bath at 80±1°C for 10min, and cool in an ice bath; add 1.25ml of solution III (mix 200ml of n-hexane and 200ml of diethyl ether evenly), shake rapidly for about 10min, discard the lower aqueous phase; add 3ml of solution IV (10.8g of sodium hydroxide dissolved in 900ml of d...

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Abstract

The invention relates to a tiny plesiomonas T7-7 for the degradation of a hydrocarbon under a low temperature and the plesiomonas application, which is characterized in that a bacterial strain is derived from a seabed mud polluted by the petroleum and is fostered and acclimatized repeatedly under 15 degree centigrade based on the unique carbon resource of a liquid wax, the bacterial strain is named as Pusillimonas sp. T7-7 and the storage number is CGMCC No. 2172; the plesiomonas is able to grow in the common beef juice, LB, nourish agar culture medium and to develop in a glucose inorganic salt culture medium and to live in a carbon resource of an alkane or a crude oil as well. The plesiomonas T7-7 has the advantages of enabling to grow on the hydrocarbon which acts as the carbon resource and provides the energy, utilizing a normal paraffin constructed by 16 carbons to 32 carbons to disintegrate the crude oil (5 g / l) with the degradation rate above 16% and to disintegrate the middle and long chain normal paraffin with the degradation rate above 40% under the temperature of 15 to 30 degree centigrade; recovering the ocean biology polluted by the ocean petroleum because a higher degradation rate is achieved when the bacterial strain is mixed and fostered with the other bacterial strain disintegrating the hydrocarbon with a surface active agent and coordinated to work on the crude oil.

Description

【Technical field】 [0001] The invention belongs to the field of microbial biotechnology and environmental biotechnology, in particular, it relates to a micromonas T7-7 (Pusillimonas sp. application. 【Background technique】 [0002] With the rapid development of industry, water pollution is becoming more and more serious, and marine pollution has become a worldwide environmental problem of concern. Among the various pollutants in the ocean, oil pollution is the most common and serious one. Petroleum is the main energy source supporting human life today, and its dominant position will hardly be shaken in the foreseeable future. Therefore, the environmental pollution caused by the storage, transportation and processing of petroleum will become increasingly serious. According to the United Nations International Maritime Organization (IMO) statistics, the world's annual oil and its products flowing into the ocean through various channels have reached 1.0×10 8 ~1.5×10 8 Ton. Of...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/31C02F3/34
Inventor 马挺李丹黄磊谢玉娟梁凤来刘如林
Owner NANKAI UNIV
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