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Mixed bacteria producing biosurfactant and screening method of mixed bacteria

A technology of surfactant and mixed bacteria is applied in the field of mixed bacteria producing biological surfactants and its screening, which can solve the problems such as the lack of breakthrough progress.

Active Publication Date: 2015-09-02
SUZHOU ZFA NEW ENERGY TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, how to screen mixed bacteria that produce biosurfactants has not yet made a breakthrough

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The culture medium involved in the present invention is specifically as follows:

[0035] 1.1 Slant medium

[0036] Beef extract 3g, peptone 10g, sodium chloride 5g, agar 20g, distilled water 1000mL, pH 7.0, steam sterilization at 121°C for 20min.

[0037] 1.2 Seed medium

[0038] Glucose 5g, beef extract 3g, peptone 10g, MgSO4*7H2O2g, deionized water 1000mL, pH7.2, steam sterilization at 121℃ for 20min.

[0039] 1.1 Starch medium

[0040] Beef extract 3g, peptone 10g, sodium chloride 5g, soluble starch 2g, agar 20g, dissolved in 1000mL of water, steam sterilized at 121°C for 20min.

[0041] 1.3 Gelatin medium

[0042] 3 g of beef extract, 10 g of peptone, and 5 g of sodium chloride, dissolved in 100 mL of water, added with 12 to 18 g of gelatin, and dissolved the above ingredients in a water bath.

[0043] 1.4 Litmus milk medium

[0044] Milk powder 100g, litmus 0.075g, dissolved in 1000mL water, pH 6.8, steam sterilized at 121°C for 15min.

[0045] 1.5 Sugar fe...

Embodiment 2

[0064] The present invention provides a method for screening mixed bacteria producing biological surfactants, comprising the following steps:

[0065] 2.1 Activation of strains

[0066] The 11 refrigerated microbial oil recovery strains were transferred to the slant medium and cultured at 37°C for 2 days.

[0067] 2.2 Seed cultivation

[0068] Take one ring of seeds activated by the inclined plane, transfer it to the Erlenmeyer flask of the seed liquid medium, and cultivate at 37° C. for 16 hours on a shaker with a rotation speed of 160 r / min.

[0069] 2.3 Fermentation culture

[0070] The seed liquid was inoculated in the initial fermentation medium (200 mL / 500 mL) with 4% inoculum, and incubated at 37° C. for 72 h on a shaker with a rotating speed of 160 r / min.

[0071] 2.4 Screening of oil recovery strains producing biosurfactants

[0072] 2.4.1 Primary screening by blood plate method

[0073] Using the hemolytic properties of biosurfactants, the strains producing bios...

Embodiment 3

[0077] The present invention provides a method for systematic classification and identification of mixed bacteria producing biosurfactants, comprising:

[0078] 3.1 Morphological identification

[0079] 3.1.1 Morphological observation of single colony

[0080] The screened strains were streaked on the plate, cultured at 37°C for 24-48h, and the colony morphology was observed.

[0081] 3.1.2 Morphological observation of bacteria

[0082] Gram staining, spore staining and capsule staining were performed on the strains, and the morphology was observed under a light microscope.

[0083] 3.2 Identification of physiological and biochemical properties

[0084] Starch hydrolysis experiment, gelatin liquefaction experiment, litmus milk experiment, sugar fermentation experiment, methyl red experiment, acetomethyl methanol experiment, citrate experiment, hydrogen sulfide experiment, contact enzyme experiment, lecithinase experiment and nitric acid were carried out on the strain Salt ...

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PUM

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Abstract

The invention relates to the microbial enhanced oil recovery technology of oil fields, and particularly relates to mixed bacteria producing a biosurfactant and a screening method of the mixed bacteria. The mixed bacteria comprise trichoderma reesei, pseudomonas stutzeri and lactobacillus rhamnosus, wherein the trichoderma reesei is purchased from the China General Microbiological Culture Collection Center (CGMCC) with the number being CGMCC No. 3.3711; the pseudomonas stutzeri is purchased from CGMCC with the number being CGMCC No. 1.10279; the lactobacillus rhamnosus is purchased from the CGMCC with the number being CGMCC No. 1.2568. The mixed bacteria are obtained by virtue of picking bacteria for oil recovery, activating and culturing a bacterial strain, primarily screening by virtue of a blood-plate method and re-screening by virtue of an oil spreading method.

Description

technical field [0001] The invention relates to an oil field microbial oil recovery technology, in particular to a mixed bacteria producing biological surfactants and a screening method thereof. Background technique [0002] Enhanced oil recovery is a type of recovery technology that enhances oil recovery by improving the physical and chemical properties of reservoirs and reservoir fluids. Oil extraction technology can be roughly divided into primary oil recovery, secondary oil recovery and tertiary oil recovery. Primary oil recovery is carried out by using the natural energy in the oil layer; secondary oil recovery is to use certain technology to restore the pressure of the oil layer to carry out oilfield development, the common one is water injection oil recovery; the subsequent technology of enhanced oil recovery belongs to tertiary oil recovery. At present, except for the Middle East, the major oil fields in the world's major oil-producing countries have entered the lat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/14C12P39/00C12R1/885C12R1/38C12R1/225
CPCC12N1/14C12N1/20C12P39/00C09K8/582C12Q1/64G01N2333/355C09K8/584G01N2333/21G01N2333/32
Inventor 段军段浩
Owner SUZHOU ZFA NEW ENERGY TECH CO LTD
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