Pseudomonas aeruginosa DB high in rhamnolipid yield and application thereof

A technology of Pseudomonas aeruginosa and rhamnolipids, which is applied in the field of microbial biotechnology and environmental biology, can solve the problems of high product cost, low yield of rhamnolipids, difficulty in large-scale industrial application, etc. The effect of low cost and simple fermentation process

Inactive Publication Date: 2016-05-11
CHINA PETROLEUM & CHEM CORP +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] As a biosurfactant, rhamnolipids are environmentally friendly, have good surface activity and emulsification function, and show great application potential in oil flooding. However, the yield of rhamnolipids is limited by the performance of strains and fermentation processes. Low lead to high product cost, it is difficult to realize its large-scale industrial application

Method used

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  • Pseudomonas aeruginosa DB high in rhamnolipid yield and application thereof
  • Pseudomonas aeruginosa DB high in rhamnolipid yield and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0039] The present invention provides the morphological characteristics, physiological and biochemical characteristics of Pseudomonas aeruginosa DBCGMMCC No. 10618 and molecular biological identification of Pseudomonas DB strain.

[0040] Such as figure 1 As shown, the morphological characteristics of Pseudomonas aeruginosa DBCGMMCC No.10618 provided by the present invention: after 1 to 2 days of culture in an aerobic solid medium, milky white colonies with a size of 1.0 to 5.0 mm generally appear, and the cell morphology is short rod-shaped. The size is 1-5μm×0.5~1.0μm. Physiological and biochemical characteristics of Pseudomonas aeruginosa DBCGMMCCNo.10618: aerobic, growth temperature 30-60°C, optimum growth temperature 30-45°C, growth pH range 4-9, optimum growth pH range 7.0-7.5, NaCl Tolerance of 0 to 5%. as attached figure 2 As shown, molecular biological identification showed that the strain was Pseudomonas aeruginosa (pseudomonasaeruginosa).

Embodiment 2

[0042] Pseudomonas aeruginosa DBCGMMCCNo.10618 culture fermentation provided by the invention and performance evaluation of fermentation broth

[0043] Seed liquid medium: glucose 10.0g / L, yeast powder 3.0g / L, KH 2 PO41.0g / L, Na 2 HPO 4 1.0g / L, MgSO 4 ·7H 2 O0.5g / L.

[0044] Fermentation medium: molasses 70g / L, waste oil 120g / L, pH value 7.0-7.5.

[0045] Fermentation conditions: inoculum size 8%, inoculation age 10h, initial pH 7.5, temperature 40°C, stirring speed 200rpm, pH=7, volume 10m 3 , Ventilation: 1.5L / min.

[0046] The concentration of rhamnolipid product detected in the fermentation broth reached 75g / L.

[0047] Performance parameter evaluation of fermentation broth:

[0048] (1) Determination of temperature resistance

[0049] Take prepared surfactant solutions with different concentrations, heat them in a thermostat to 150°C for two hours, then measure their (table) interfacial tension at room temperature or 60°C.

[0050] (2) Determination of salt tolera...

Embodiment 3

[0064] Example 3 Application of Pseudomonas DB Fermented Liquid in Field Oil Displacement of Block A in Shengli Oilfield

[0065] The application of the fermented liquid provided by the present invention in the Shengli Oilfield block A block overview: formation temperature 90°C, formation water salinity 7.1×10 4 mg / L, geological reserves 3.5×10 4 t, pore volume 3.5×10 4 m 3 .

[0066] Implementation steps: the rhamnolipid fermentation liquid and the oil field injection water are mixed according to the volume ratio of 1:1000 and then injected into the oil reservoir. The total amount of mixed liquid injected is 25% of the pore volume of the oil reservoir, which is 0.875×10 4 m 3 , with an injection velocity of 100m 3 / d, the injection method is slug type.

[0067] Test results: After the test is completed, the block has accumulatively increased oil production by 4,200 tons and enhanced recovery by 12.0%.

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Abstract

Pseudomonas aeruginosa DB high in rhamnolipid yield and application thereof are provided. This strain of the invention is obtained by subjecting an oil-water well sample from Shengli Oil Field to blood plate and oil spreader ring screening, is obtained by repeated screening and passage through temperature resistance and salt resistance investigation and is named Pseudomonas aeruginosa DB, collected under CGMMCC No. 10618. A seed solution medium for the Pseudomonas aeruginosa DB comprises glucose 10.0 g / L, yeast powder 3.0 g / L, KH2PO4 1.0 g / L, Na2HPO4 1.0 g / L, and MgSO4.7H2O 0.5 g / L; a fermentation medium for the Pseudomonas aeruginosa DB comprises molasses 70 g / L and a waste oil 120 g / L and has a pH value of 7.0-7.5; growth temperature is 30-60 DEG C, optimal growth temperature is 40-45 DEG C, growth pH range is 4-9, optimal growth pH range is 7.0-7.5, and NaCl tolerance is 0-5 %. A physical simulation test shows an increase of higher than 13 % in recovery rate, a field test shows an increase of higher than 8 % in recovery rate; therefore, the Pseudomonas aeruginosa DB high in rhamnolipid yield provided by the invention is widely applicable to the field test for increasing crude oil recovery rate.

Description

technical field [0001] The invention belongs to the field of microbial biotechnology and environmental biotechnology, and in particular relates to a high-yield rhamnolipid Pseudomonasaeruginosa.DB strain and its application. technical background [0002] In the case of increasingly tense energy resources, enhanced oil recovery has become the focus of oil exploration research. In recent years, tertiary oil recovery technology has been widely valued and field applied. Therefore, the research and development of tertiary oil recovery oil displacement agents has also increasingly attracted scientific research Due to the concerns of the researchers, the commonly used oil displacement agents are mainly chemical surfactants, such as petroleum sulfonates, alkylbenzene sulfonates and petroleum carboxylates. At present, chemical surfactants are the main oil displacement agents in oil production, but due to the poor temperature and salt resistance of chemical surfactants and poor adapta...

Claims

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Application Information

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IPC IPC(8): C12N1/20C09K8/582C12R1/385
Inventor 林军章冯云谭晓明段传慧王静徐登霆孙刚正胡婧刘涛巴燕宋永亭
Owner CHINA PETROLEUM & CHEM CORP
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