Method for extracting and separating proteins from cake by using ionic liquid and enzyme process

An ionic liquid and enzymatic extraction technology, applied in the field of protein separation and purification, can solve the problems of complex operation, high cost and low extraction rate.

Inactive Publication Date: 2012-11-28
NANJING UNIV OF FINANCE & ECONOMICS
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The traditional method of extracting and separating proteins still has the disadvantages of low extraction rate, high cost and complicated operation, so finding an efficient, green and economical method for extracting and separating proteins has become the focus of technical research

Method used

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  • Method for extracting and separating proteins from cake by using ionic liquid and enzyme process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] (1) Crush and sieve high-temperature rapeseed meal, degrease petroleum ether to obtain defatted rapeseed meal powder;

[0042] (2) Mix defatted rapeseed meal powder with phosphate buffer solution with pH=8.0, add α-amylase and cellulase to make a mixture, enzymatically hydrolyze the mixture at 60 °C for 4 hours, and then 4000 r min -1 Centrifuge for 20 minutes to obtain enzymatic hydrolysis solution and precipitate.

[0043] The solid-liquid mass ratio of the defatted rapeseed meal powder and the phosphate buffer is 1:15;

[0044] The enzyme activities of α-amylase and cellulase in the mixed solution were 110 U·g -1 and 60 U·g -1 , with a total enzyme activity of 170 U·g -1 .

[0045] (3) Add the precipitate from step (2) to 10 times the amount of water, and use 0.5 mol L -1 NaOH was adjusted to pH=10 for alkali dissolution reaction, after 2h, 4000 r·min -1 Centrifuge for 20min, combine the separated supernatant with the enzymolysis solution in step (2), add 0...

Embodiment 2

[0050] (1) crushing and sieving high-temperature rapeseed meal, degreasing with cyclohexane to obtain defatted rapeseed meal powder;

[0051] (2) Mix defatted rapeseed meal powder with phosphate buffer solution at pH = 8.0, add α-amylase and cellulase to make a mixture, enzymatically hydrolyze the mixture at 55 °C for 3 h, and 4000 r· min -1 Centrifuge for 20 minutes to obtain enzymatic hydrolysis solution and precipitate.

[0052] The solid-to-liquid mass ratio of the defatted rapeseed meal powder to the phosphate buffer is 1:10;

[0053] The enzyme activities of α-amylase and cellulase in the mixed solution were 90 U·g -1 and 50 U·g -1 , with a total enzyme activity of 140 U·g -1 .

[0054] (3) Add the precipitate from step (2) to 10 times the amount of water, and use 0.5 mol L -1 NaOH was adjusted to pH=10 for alkali dissolution reaction, after 2h, 4000 r·min -1 Centrifuge for 20 min, combine the separated supernatant with the enzymatic hydrolysis solution in step ...

Embodiment 3

[0059] (1) crushing and sieving low-temperature rapeseed meal, degreasing with n-hexane to obtain defatted rapeseed meal powder;

[0060] (2) Mix defatted rapeseed meal powder with boric acid buffer solution at pH=9.0, add α-amylase, cellulase and pectinase to make a mixture, and enzymolyze the mixture at 60 °C for 5 h , 4000 rpm -1 Centrifuge for 20 minutes to obtain enzymatic hydrolysis solution and precipitate.

[0061] The solid-liquid mass ratio of the defatted rapeseed meal powder and boric acid buffer solution is 1:15;

[0062] The enzyme activities of α-amylase, cellulase and pectinase in the mixed solution were 70 U·g -1 , 40 U·g -1 and 50 U·g -1 , with a total enzyme activity of 160 U·g -1 .

[0063] (3) Add the precipitate from step (2) to 10 times the amount of water, and use 0.5 mol L -1 NaOH was adjusted to pH=10 for alkali dissolution reaction, after 2h, 4000 r·min -1 Centrifuge for 20min, combine the separated supernatant with the enzymolysis solut...

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Abstract

The invention relates to a field of separation and purification of proteins, in particular to a novel method for extracting and separating proteins from cake by using an ionic liquid and an enzyme process. The method for extracting and separating proteins from cake by using an ionic liquid and an enzyme process disclosed by the invention comprises the following steps of: (1) smashing and sieving cake serving as a raw material, and degreasing to obtain degreased cake powder; (2) performing enzymolysis on the degreased cake powder under the condition that the pH is 7-10, the temperature is 40-65 DEG C and the material to liquid ratio is 1:7-1:15 in a state that the total enzyme activity is 90-250 U.g<-1> for 1-5 hours, and centrifugally separating; (3) performing alkali-solution and acid-isolation on the obtained precipitate and hydrolysate, and performing freeze drying to obtain crude proteins; (4) adding the crude proteins obtained in the step (3) into an ionic liquid/salt dual-water-phase system, and separating and purifying proteins at the temperature of 40-70 DEG C and at the pH of 4-9; and (5) separating proteins from an ionic liquid phase obtained in the step (4) with a membrane separation method, recovering an ionic liquid, and performing freeze drying to obtain high-purity protein powder. The method is easy to operate, is environment-friendly, and is a novel environment-friendly, energy-saving and efficient protein separating and purifying method; and the protein purity is high, and the bioactivity is kept.

Description

technical field [0001] The invention relates to the field of protein separation and purification, in particular to a method for extracting and separating protein in cake by using ionic liquid and enzymatic method. Background technique [0002] Oilseed cakes generally contain protein, polyphenols, phytic acid, polysaccharides and other components with high utilization value. Cake is a plant protein resource with high potential nutritional value, and its protein content is 35%~40%, which is the most valuable component in cake. This protein contains a large number of essential amino acids needed by the human body, so it is also an extremely important food protein source. Here we take rapeseed protein in rapeseed meal as an example. The total amount of amino acids in rapeseed meal accounts for 83.8% of its total protein, and its lysine content is close to that of soybean, while its methionine content is higher than that of soybean. Used to supplement grains and many legumes t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14
CPCY02P20/10Y02P20/50
Inventor 刘晓庚刘琴陈梅梅潘春淋高梅万忠民曹崇江彭冬梅王立峰吴珂
Owner NANJING UNIV OF FINANCE & ECONOMICS
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