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Method for separating and purifying hydrophobin

A technology of hydrophobin and purification method, which is applied in the field of separation and purification of hydrophobin, can solve problems such as separation and purification difficulties, hinder development speed, and limited production capacity of hydrophobin, and achieve broad application prospects, easy industrial application, and low cost.

Inactive Publication Date: 2007-10-17
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the production capacity of hydrophobin is still very limited, and there are also great difficulties in separation and purification, which greatly hinders its development speed.

Method used

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  • Method for separating and purifying hydrophobin
  • Method for separating and purifying hydrophobin
  • Method for separating and purifying hydrophobin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1. Separation and purification of hydrophobin and detection of the influence of ultrasonic oscillation treatment with different powers on the purification effect

[0022] 1. Preparation of solid substrate with Teflon particles distributed on the surface for separation and purification of hydrophobin

[0023] Put the clean polished quartz substrate into the ion beam sputtering coating machine, install the Teflon target, vacuumize to 1.3E-3pa, take it out after sputtering for 5s, and obtain a solid substrate with Teflon particles distributed on the surface. The observation results of the atomic force microscope are shown in the figure 1 (the size of Figure A is 5 microns, and the size of Figure B is 1 micron), the surface roughness of the clean and polished quartz substrate is very low, and it can be seen from the atomic force image after sputtering that there are many particles distributed on the surface of the substrate. Particles, through the test of the water ...

Embodiment 2

[0026] Embodiment 2, the separation and purification of hydrophobin

[0027] 1. Preparation of solid substrate with Teflon particles distributed on the surface for separation and purification of hydrophobin

[0028] Put the clean glass substrate into the ion beam sputtering coating machine, install the Teflon target, vacuumize to 1.3E-3Pa, take it out after sputtering for 300s, and obtain a solid substrate with Teflon particles distributed on the surface.

[0029] 2. Separation and purification of hydrophobin

[0030] Put the solid substrate with Teflon particles distributed on the surface prepared in step 1 in the hydrophobin HFBII solution to be purified and soak for 72 hours, then take the substrate out of the hydrophobin solution, observe and analyze the image under an atomic force microscope, you can see Large irregular-shaped hydrophobin aggregates grew on the surface of the substrate, which was caused by the adsorption and nucleation of hydrophobin on the surface of Te...

Embodiment 3

[0031] Embodiment 3, the separation and purification of hydrophobin

[0032] 1. Preparation of solid substrate with Teflon particles distributed on the surface for separation and purification of hydrophobin

[0033] Put the clean glass substrate into the ion beam sputtering coating machine, install the Teflon target, vacuumize to 1.3E-3Pa, take it out after sputtering for 600s, and obtain a solid substrate with Teflon particles distributed on the surface.

[0034] 2. Separation and purification of hydrophobin

[0035] Soak the solid substrate with Teflon particles on the surface prepared in step 1 in the hydrophobin HFBII solution to be purified for 24 hours, then take the substrate out of the hydrophobin solution, observe and analyze the image under an atomic force microscope, you can see Large irregular-shaped hydrophobin aggregates grew on the surface of the substrate, which was caused by the adsorption and nucleation of hydrophobin on the surface of Teflon particles. The...

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Abstract

The invention discloses hydrophobic protein separation and purification method. The method comprise: soaking solid substrate with hydrophobic particle on surface in hydrophobic protein solution, then taking the substrate out of the hydrophobic protein solution, rinsing, carrying out supersonic oscillation in container filled with water, then taking the substrate out of the solution, obtaining purified hydrophobic protein solution. The principle that hydrophobic protein can adsorb at hydrophobic substance surface, and hydrophobic particle has hetero phase nucleation growth effect to hydrophobic protein can be utilized to achieve the aim for separating and purifying the hydrophobic protein. The inventive method is simply and conveniently operated, and cost is low. The hydrophobic substance surface can be repeatedly used which is a merit for industrial production. The present invention will have a broad prospect in hydrophobic protein separation and purification field.

Description

technical field [0001] The invention relates to a method for separating and purifying proteins, in particular to a method for separating and purifying hydrophobin. Background technique [0002] Fungal hydrophobin is a cluster of small proteins (about 100 amino acid residues) with unusual physiological and biochemical properties widely present in filamentous fungi, which have not been found in other organisms. Based on the hydrophobicity profile and the solubility characteristics of their assembled films, hydrophobins can be divided into two classes: type I and type II. In order to understand hydrophobin more concretely, scientists have done a lot of research on their structure and biophysical and biochemical characteristics. Studies have found that hydrophobin plays a very important role in the growth and reproduction of fungi, including the adsorption and stability of fungi to surfaces, the formation of pulp and aerial hyphae, and the covering and protection of fungal stru...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14
Inventor 胡振彦李良彬田扬超
Owner UNIV OF SCI & TECH OF CHINA
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