Protein with emulsion ability and application thereof
An emulsifying activity and protein technology, applied in the field of proteins with emulsifying activity, can solve the problems of difficult to fully meet the repair requirements, slow recovery process efficiency, secondary pollution, etc. effect, cost saving effect
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Embodiment 1
[0022] Example 1: Screening and sequence analysis of emulsified proteins
[0023] The applicant screened and obtained a strain of Bacillus licheniformis XS2-450 (Geobacillus sp.), and found that its fermentation product had emulsifying activity. The preservation number of the strain is CGMCCNo.9430, and it was preserved on July 09, 2014 at the General Microbiology Center of the Chinese Microbiological Culture Collection Management Committee, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.
[0024] The protein with emulsifying effect of the present invention is obtained by purifying the metabolites of the bacterial strain. 1) Preparation of crude extract
[0025] The active strain Geobacillus sp.XS2-450 was shaken at 60°C and 180rpm for 10 hours as a seed solution for fermentation, and the inoculation amount was 10% (v / v), inoculated into the fermentation medium, and the final concentration of 0.1 % palm oil is used as the inducer of producing highly active biol...
Embodiment 2
[0042] This example illustrates the method for recombinant expression and activity verification of a protein with a molecular weight of 24K Da.
[0043] 1. Construction of recombinant plasmid vector
[0044] (1) Amplification of the target gene
[0045] Use the nucleotide sequence corresponding to the 24K protein in Example 3 and Primer Premier 5.0 to carry out primer design
[0046]
[0047] The genome of the active strain Geobacillus sp.XS2-450 was used as a template for PCR amplification.
[0048] PCR reaction system:
[0049]
[0050] The reaction conditions are:
[0051] 95°C for 15min (95°C for 30s, 59°C for 30s, 72°C for 2min)·30cycles; 72°C for 5min. The PCR amplified product was recovered from the column.
[0052] (2) Enzyme digestion of target gene and pET21a+ vector
[0053] The recovered fragment and the pET21a+ vector were digested with fastdigestNdeI and fastdigestHindIII respectively.
[0054] Enzyme digestion system and conditions:
[0055]
[...
Embodiment 3
[0083] Example 3 Application of emulsifier protein in oil sludge disposal.
[0084] The present invention mainly utilizes the emulsifier protein prepared in Example 2 to treat oil sludge polluted by high viscosity and heavy hydrocarbons.
[0085] Stir the sludge to make it homogeneous, adjust the ratio of sludge and emulsifier protein to 4:3, add water 10 times that of the sludge, stir fully at 60°C to emulsify the crude oil in the sludge, and let it stand , The settled oil residue is deeply deoiled to remove residual high-viscosity components, so as to achieve the purpose of separating the oil and residue in the sludge, and finally recover all the emulsified upper crude oil. During the treatment process, the oil content of the sludge is monitored, and the oil content in the sludge has been reduced from the original 6.1% to the current 0.8%, and the oil content has been reduced by more than 87%, which meets the sludge disposal standard; compared with physical and chemical trea...
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