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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

Active Publication Date: 2017-04-26
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the existing restoration technology and oil recovery process, there are certain limitations in physical and chemical restoration, and it is difficult to fully meet the restoration requirements, while the recovery process has problems such as slow efficiency and serious secondary pollution.

Method used

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  • Protein with emulsion ability and application thereof
  • Protein with emulsion ability and application thereof
  • Protein with emulsion ability and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention provides a protein with the emulsion ability and application thereof. The amino acid sequence of the emulsified protein is SEQ ID NO:1. The emulsified protein is used for processing high-viscosity oil sludge contaminated by heavy hydrocarbon compounds. When processing the high-viscosity oil sludge contaminated by the heavy hydrocarbon compounds, the obtained emulsified protein has a very good deoiling effect and can enable the content of oil in the oil sludge to be reduced to now 0.8% from originally 6.1%, and the oil content is reduced by 87%. Compared with a physical or chemical method, the cost can be saved by 30%, the protein exhibits great economic benefits and scaled application potentialities while cost saving and environmental protection are achieved.

Description

technical field [0001] The invention belongs to the technical field of bioactive molecular screening, and in particular relates to a protein with emulsifying activity and its application. Background technique [0002] Bioemulsifier is a biological macromolecular compound that exhibits important physiological and metabolic activities in the life process; at the same time, studies have shown that the proportion of protein in emulsifiers varies greatly, but its content has no significant relationship with emulsifying activity. The hydrophobic amino acid sequence is the active site for emulsification. The position and number of hydrophobic amino acids play a great role in the emulsification activity. It is clear that the emulsification activity is closely related to the molecular structure of the emulsifier, especially It is the change of protein structure that has a great influence on the emulsifying activity. The protein is connected to the sugar structure at one end through ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/32C12N15/31C02F11/02C02F101/32
CPCC02F11/02C02F2101/32C07K14/32
Inventor 王兴彪高小龙黄志勇董大媛
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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