Brevibacillus agri., preparation thereof, method for preparing surfactant and uses of brevibacillus agri. and surfactant
A technology of brevibacillus and active agent, applied in the field of brevibacillus and preparation and preparation of surfactants, to achieve good emulsification performance, reduce surface tension, and good physical properties
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Embodiment 1
[0055] Embodiment 1: Isolation, identification and preservation of biosurfactant bacterial classification
[0056] 1. Enrichment culture and isolation of surfactant-producing strains
[0057] According to the conventional strain screening method, 10 mL of oil and water samples collected from oil fields in western China were inserted into 100 mL of sterilized crude oil medium (2% crude oil, v: v), 30 ° C, 180 rpm constant temperature shaking culture for 96 h, and then Take 5% of the enriched culture solution of the experimental group with a high degree of emulsification and dispersion of crude oil and transfer it to a fresh medium, and culture it at 30° C. and 180 rpm for 96 hours with constant temperature shaking. Repeat the enrichment culture for 10 rounds, then select the experimental group with the best crude oil emulsification and dispersion, take 100 μL of fermentation liquid and spread it on the LB agar plate medium, and cultivate it at 30°C for 48 hours; pick single col...
Embodiment 2
[0071] Embodiment 2: Extraction of B5-2 metabolites
[0072] 1. Fermentation of B5-2
[0073] For the Brevibacillus Agri. B5-2 provided by the present embodiment, the fermentation medium comprises: MgSO 4 0.2g / L, K 2 HPO 4 1.0 g / L, KH 2 PO 4 1.0 g / L, Na 2 HPO 4 4.0 g / L, NaCl 10.0 g / L, NaNO 3 10.0 g / L, crude oil (heavy oil) 10.0g / L, molasses 2-3g / L, pH 7.0, appropriate amount of trace elements and multivitamin solution, pH 6-8, sterilized at 115°C for 30min, and fermented at 30°C.
[0074] Use an inoculation loop to streak the preserved strains on the plate to activate them, culture them at a constant temperature of 30°C for 20 hours, and then pick three rings (each inoculation loop contains more than 3 single colonies with obvious characteristics) strains from the plate to inoculate into the first level Seed shake flask (250mL Erlenmeyer flask, liquid volume is 50mL), cultured at 30°C, 180r / min for 18h, 2% of the volume was inserted into the secondary shake flask (250...
Embodiment 3
[0086] Example 3: Optimization of B5-2 lipopeptide-producing culture system
[0087] Culture medium includes seed medium, fermentation medium and slant medium, wherein:
[0088] Seed medium (g / L): MgSO 4 0.2,K 2 HPO 4 1.0, KH 2 PO 4 1.0, NH 4 NO 3 1.0, yeast extract 1.0, CaCL 2 2H 2 O 0.02, FeCl 3 0.05, distilled water 1000ml, PH 7.2;
[0089] Fermentation medium (g / L): MgSO 4 0.2,K 2 HPO 4 1.0, KH 2 PO 4 1.0, Na 2HPO 4 4.0, NaCl 10.0, peptone 10.0, crude oil 20.0, molasses 10.0, distilled water 1000ml, PH 7.0;
[0090] Incline medium (g / L): beef extract 5.0, peptone 10.0, NaCl 5.0, agar 20.0, distilled water 1000ml, pH 7.0.
[0091] 1. Carbon source optimization
[0092] Any nutrient that can be used as a source of carbon in microbial cell structure or metabolites is called a carbon source. There are a wide variety of carbon sources that can be used as microbial nutrition, ranging from simple inorganic substances (CO 2 , carbonate) to complex organic carbo...
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