Reverse micelle system used for extracting proteins, and preparation method and application thereof

A technology of reverse micelles and proteins, applied in the field of bioengineering, to achieve good biodegradability, biocompatibility, and high extraction rate

Inactive Publication Date: 2018-01-19
DALIAN POLYTECHNIC UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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  • Reverse micelle system used for extracting proteins, and preparation method and application thereof
  • Reverse micelle system used for extracting proteins, and preparation method and application thereof
  • Reverse micelle system used for extracting proteins, and preparation method and application thereof

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[0032] Example 1

[0033] The starch laurate reverse micelle system used for protein extraction was prepared according to the following method:

[0034] 1. Synthesis of Starch Lauric Acid

[0035] (1) Add 1mol / L hydrochloric acid to corn starch to keep the pH of the solution at 3-5, stir at 60℃ for 1-4h, filter, and wash the filter cake with distilled water until the pH is about 7, and the average molecular weight is 20000-40000 degraded corn starch;

[0036] (2) Take 6 g of degraded corn starch in a three-necked flask, add 0.3 g of LiCl, fill the reaction system with nitrogen to replace the air in the system to achieve an anhydrous environment, add 50 mL of NN dimethylacetamide, and heat to 80°C. Stir for 30min, add different proportions of pyridine (0.019-0.111moL) and lauroyl chloride (0.019-0.111moL) to the reaction system. After reacting for 30min, cool to room temperature, add proper amount of ethanol, precipitate, filter by suction, deionized water, anhydrous Washed with etha...

Example Embodiment

[0045] Example 2

[0046] Soy protein extracted with starch laurate reverse micelle system:

[0047] For the reverse micelle system (reverse micelle system 3) formed by the above-prepared lauroyl substitution degree of 1.93 starch laurate, add distilled water to form a reverse micelle system with a concentration of 1.0 to 4.0 mg / mL. Add 0.015g of soy protein powder to the mixture, shake at 35℃, 100r / min for 30min, centrifuge at 8000r / min for 20min, the extraction system will be divided into two layers, the upper layer is the reverse micelle system for protein extraction, and the lower layer is the residue. The residue was removed by filtration, and the supernatant was taken to measure the absorbance A at 280 nm with an ultraviolet-visible spectrophotometer. Take bovine serum albumin as the standard product, make the standard curve, the linear regression equation is: A=0.666C-0.0025, R 2 =0.9999; where A is the absorbance and C is the protein concentration (mg / mL).

[0048] Bring th...

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Abstract

The invention provides a reverse micelle system used for extracting proteins, and a preparation method and an application thereof. The reverse micelle system is composed of hydrophobic starch and a nonpolar organic solvent, wherein substitution degree of the hydrophobic starch is 0.5-3; the nonpolar organic solvent includes isooctane and n-octanol according to the volume ratio of 1:1-9:1; the mass-volume ratio (g:mL) of the hydrophobic starch to the nonpolar organic solvent is 0.001:1-0.01:1. A surfactant for preparing a reverse micelle solution is a starch-based product, which has excellent biodegradability and biocompatibility. The reverse micelle system has higher extraction ratio to macromolecular proteins than other reverse micelle systems.

Description

technical field [0001] The invention belongs to the technical field of bioengineering and relates to a reverse micelle system, in particular to a reverse micelle system for extracting protein and its preparation method and application. Background technique [0002] Reverse micelles are based on dipole-dipole, electrostatic force, hydrophobic, etc. When the concentration of amphiphilic substances in non-polar organic solvents exceeds the critical aggregation concentration, reverse aggregates will be formed in the organic phase , such as reverse micelles and other forms. The non-polar groups of the reverse micelles face outward, and the interior is a hydrophilic inner core, which can solubilize biologically active components such as proteins. Due to the shielding protection of reverse micelles, these substances can be kept from direct contact with organic solvents to protect the activity of biological substances, thereby realizing the dissolution and separation of certain bio...

Claims

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Application Information

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IPC IPC(8): C07K1/14
Inventor 汪源浩徐成张红奎谭凤芝李沅曹亚峰
Owner DALIAN POLYTECHNIC UNIVERSITY
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