A Pseudomonas stutzeri producing bioemulsifier
A technology of Pseudomonas stutzeri and emulsifier, applied in the biological field, can solve problems such as disadvantage, narrow temperature resistance, acid and alkali resistance range, poor diesel emulsification effect, etc.
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Embodiment 1
[0034] This example illustrates the method for obtaining the strain NJtech 11-1 of the present invention.
[0035] A ring of bacteria was picked from the slant of miscellaneous bacteria provided by Shengli Oilfield, inserted into LB liquid medium, and cultivated at 37° C. and 200 rpm for 12 hours to obtain a seed solution. Use a pipette gun to take 1 mL of the seed solution in the ultra-clean workbench and put it into a sterile test tube of No. 1 9 mL sterile water, and shake it fully to make the bacterial solution evenly dispersed, which is 10 -1 sample diluent; take another 10 -1 1 mL of sample diluent in No. 2 sterile test tube of 9 mL sterile water, and so on, mark 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 and 10 -8 . Take 10 respectively -7 and 10 -8 Dilution 200 μL of the dilution solution is evenly coated on the plate with a sterile coating stick, and left to stand for 20 minutes, then inverted and placed in a 37°C constant temperature incubator for 12-24 hours...
Embodiment 2
[0039] This example illustrates the method for the production of emulsifier by the strain NJtech 11-1 provided by the present invention using glucose as a carbon source.
[0040] The bacterial strain NJtech 11-1 stored in a glycerol tube was activated, spread on an LB plate, and cultured at 37°C for 12 hours. Pick a single colony and inoculate it into LB liquid medium, culture at 37°C, 200rpm, for 12 hours. Then inoculate the emulsifier-producing fermentation medium with 2% inoculum amount, and cultivate for 48 hours at 37° C. and 200 rpm.
[0041] The formula of the fermentation medium for producing emulsifier is: glucose 20g / L, yeast extract 0.5g / L, sodium nitrate 4g / L, dipotassium hydrogen phosphate 2g / L, potassium dihydrogen phosphate 2g / L, calcium chloride 0.01g / L, magnesium sulfate heptahydrate 0.3g / L. Adjust tap water to 1000mL. The pH is 7.4.
Embodiment 3
[0043]The fermented liquid cultured in Example 2 was centrifuged at 4° C., 8000 rpm for 20 minutes, and the supernatant was collected. A part of the fermented supernatant collected is directly used for the determination of emulsifying activity; another part of the fermented supernatant collected is used for the extraction of the emulsifier, the specific method is as follows: pre-cool the supernatant of the fermented liquid, and then Add 3 times the volume of pre-cooled acetone and place in the refrigerator overnight. Then it was centrifuged at 8000rpm at 4°C for 10 minutes, the precipitate was collected, dissolved with an appropriate amount of distilled water, dialyzed at 4°C for 48h, freeze-dried, and weighed.
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