Bacillus amyloliquefaciens for producing surfactin, and application thereof
A technology of amylolytic spores and surfactin, which is applied in the field of microorganisms, can solve problems such as the lack of amyloliquefaciens, and achieve the effects of high surface activity, high-efficiency crude oil and guar gum degradation ability, and high crude oil emulsification and viscosity-reducing ability
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[0038] Example 1 Screening and identification of strain CB-019
[0039] 1. Isolation of strain CB-019
[0040] (1) Sample source: produced fluid from an oil field.
[0041] (2) Preliminary strain screening: Dilute the sample with sterile water in a concentration gradient to 10 -3 ~10 -8 , Take 0.1mL evenly spread on the blood plate culture medium, put it in a 37℃ incubator for culture. Transfer the colonies with the hemolysis circle to a fresh blood plate, inoculate it into the slant medium after streaking and purification, and store at 4°C for later use.
[0042] (3) Re-screening of strains: inoculate the preserved strains into LB medium, culture with shaking at 37°C and 160rpm for 24h, then inoculate the fermentation medium with an inoculum of 1:50, and cultivate 30 with shaking at 37°C and 160rpm ~36h. Take 30 mL of the above fermentation broth and measure the surface tension of the supernatant after centrifugation at 5000 r / min. The strain with the lowest surface tension value ...
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[0048] Example 2 Extraction and identification of fermentation products of strain CB-019
[0049] Fermentation of strain CB-019: Inoculate the Bacillus amyloliquefaciens CB-019 strain on LB solid medium, culture at 35°C for 1d, inoculate a single colony into the seed medium, culture at 35°C, 150rpm with shaking for 2d , Into the seed liquid, the seed liquid is connected to the fermentation medium according to the volume ratio of 1:20, shake culture at 35° C. and 150 rpm for 3 days, and centrifuge to obtain the fermentation liquid.
[0050] The fermentation broth is extracted by acid precipitation method to extract the fermentation products in CB-019: the fermentation broth is centrifuged to remove the bacteria, the supernatant pH is adjusted to 2.0, and the supernatant is allowed to stand overnight. The precipitate is collected by centrifugation. The precipitate is washed with pH 2.0 hydrochloric acid and NaHCO 3 The powder adjusts the hydrochloric acid washing solution to pH 8.0, a...
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[0054] Example 3 Determination of the biologically active substance performance of strain CB-019
[0055] The fermentation broth of strain CB-019 obtained in Example 2 was used for the following determinations.
[0056] 1. Determination of emulsification performance
[0057] (1) Fermentation liquid drain ring measurement: 0.5g Sudan Red III is added to 100mL liquid paraffin, the liquid paraffin is dyed, filtered and then used. Add 8mL of the dyed liquid paraffin to a 9cm dish filled with 60mL of distilled water. After a uniformly dispersed dyed liquid paraffin film is formed on the surface of the dish, slowly drip 1mL of the fermentation broth of strain CB-019 in the center of the oil film, and observe The size of the drain ring, the result is Figure 5 As shown, the results show that CB-019 can produce a larger oil drain ring with a diameter of about 5-6cm.
[0058] (2) Emulsification rate of kerosene and liquid paraffin: Take 5 mL of kerosene and liquid paraffin, and mix them with ...
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