Preparation method and application of protein nanocage stabilized Pickering emulsion

A nanocage and protein technology, applied in the preparation method, application, chemical instrument and method of peptides, etc., can solve the problems of structural damage, loss of biological activity, etc., achieve strong biocompatibility, ensure stability, and simple preparation process Effect

Active Publication Date: 2019-04-26
QINGDAO GUGAO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Unsuitable selection methods may lead to structural disru

Method used

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  • Preparation method and application of protein nanocage stabilized Pickering emulsion
  • Preparation method and application of protein nanocage stabilized Pickering emulsion
  • Preparation method and application of protein nanocage stabilized Pickering emulsion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation method of GroEL nanocage-stabilized Pickering emulsion

[0026] The basic principle of Pickering emulsion stabilized by GroEL nanocages is as follows: figure 1 shown.

[0027] (1) Using the GroEL nanocage as an emulsifier, the expression and purification steps of GroEL are as follows:

[0028] The pTric99 plasmid containing the GroEL gene was transformed into Escherichia coli strain BL21 (DE3); liquid medium was used for the selective amplification of the transformed plasmid BL21 (DE3), when OD 600 When the value reached 0.4-0.6, the final concentration of 1mM isopropyl-β-D-1-thiogalactopyranoside was used to induce BL21(DE3) to express protein at 37°C; after 4 hours of induced expression, the bacteria were collected body, resuspension and cell disruption in a high-pressure cell disruptor; the precipitation and crude extraction of GroEL were carried out using a saturated ammonium sulfate solution with a final concentration of 50%; anion exchange ...

Embodiment 2

[0036] Embodiment 2 Pickering emulsion stability test

[0037] (1) pH value: all emulsions are prepared at neutral pH, and the pH value of the newly prepared emulsion can be changed by adding hydrochloric acid or sodium hydroxide. Systematic research shows that the emulsion is very stable in the neutral to alkaline range, and the pH value of the emulsion should not be lower than 4.5, and should not be higher than 10.5.

[0038] (2) Ionic strength: The surface charge is an important factor for the stability of the Pickering emulsion, and the surface charge changes with the change of the ionic strength. Systematic research shows that when the ionic strength is not higher than 200mM, the stability of the formed emulsion is better.

[0039] (3) Storage temperature: the Pickering emulsion was incubated at different temperatures for several days, and the influence of the analysis temperature on the Pickering emulsion was investigated. At the upper limit temperature of 65°C, the Gr...

Embodiment 3

[0040] Example 3 Application of GroEL Nanocage Stabilized Pickering Emulsion

[0041] a. Prepare a GroEL buffer solution (50 mM Tris-HCl, 1 mM EDTA, 1 mM DTT, 0.02% sodium azide, pH=7.5), and control the mass fraction of GroEL at 0.25 (wt%).

[0042] b. First dissolve β-carotene into the oil phase substance, and then mix the oil phase substance dissolved with β-carotene and water in a ratio of 0.11:1 by volume.

[0043] c. adding the GroEL solution obtained in step a dropwise to the oil and water mixture obtained in step b while stirring the mixture, thereby preparing a heterogeneous coarse emulsion with a large droplet diameter.

[0044] d. Utilize the KQ-100KDE high-power numerically controlled ultrasonic instrument to ultrasonically treat the heterogeneous coarse emulsion obtained in step c for 2 minutes with an amplitude of 40%, so as to homogenize the coarse emulsion, thereby obtaining a stable oil-in-water Pickering emulsion .

[0045] In this example, β-carotene was u...

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Abstract

The invention provides a preparation method of protein nanocage stabilized Pickering emulsion in order to overcome defects that nano-particle stabilized Pickering emulsion is poor in biocompatibilityand biodegradability and common bioorganic molecules are poor in surface activity, have insufficient monodispersity and wetting performance and are easily affected by environmental conditions, and belongs to the field of Pickering emulsion. According to a specific scheme, a buffer solution of a GroEL protein nanocage is prepared, and mass fraction of GroEL is controlled to be 0.05wt%-0.45wt%; an oil-phase substance and water are mixed in a certain proportion, and the range of a volume ratio of the oil-phase substance to water is controlled to be 0.11-0.50; the GroEL solution is added to an oil-water mixture dropwise, meanwhile, the mixture is stirred, ultrasonic treatment is performed, and crude emulsion is enabled to be homogenized. The invention further provides an application of the Pickering emulsion as a bioactive substance for carrier encapsulating or transport. The Pickering emulsion prepared on the basis of the unique structure and excellent interfacial activity of GroEL has the advantages of good stability, long quality guarantee period, high biocompatibility and simple preparation process and is environmentally friendly.

Description

technical field [0001] The invention belongs to the field of Pickering emulsion preparation, and in particular relates to a preparation method and application of a protein nanocage-stabilized Pickering emulsion. Background technique [0002] Pickering emulsion is a new type of emulsion that replaces the traditional organic surfactant stable emulsion system with solid particles. It is used in a wide range of fields, including oil flooding, food, cosmetics, biomedicine, fine chemical synthesis, etc. Various inorganic particles such as silica, palygorskite, clay, etc. can be used for the stabilization of Pickering emulsions, but their application in human life is greatly limited due to their poor biocompatibility and biodegradability. In recent years, the stability of Pickering emulsions has been extended from the use of inorganic particles to the use of organic particles, especially organic particles of biological origin, such as cellulose, modified starch, flavonoids, lipid n...

Claims

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Application Information

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IPC IPC(8): C08J3/03C12N15/70C07K14/245C07K1/36C07K1/30C07K1/18C07K1/16A23L33/105A23L33/195C08L89/00
CPCA23V2002/00A23L33/105A23L33/195C07K14/245C08J3/03C08J2389/00C12N15/70A23V2250/546A23V2250/211
Inventor 王小强王华高娟郝锰
Owner QINGDAO GUGAO BIOTECH CO LTD
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