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Method for quick separation and purification of walnut protein from walnuts

A technology for separation and purification of walnut protein, which is applied in the field of rapid separation and purification of walnut protein, can solve the problems of inability to obtain pure walnut protein, uneven specific surface area of ​​particles, and decline in quality of walnut protein, so as to speed up the separation and purification speed, and the process is simple and feasible, Reduce the effect of denaturation inactivation

Pending Publication Date: 2020-01-03
ZHEJIANG LIZIYUAN FOOD CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese patent CN103535508A discloses walnut protein powder and its preparation method. The method undergoes pressing and other processes. Although the oil content is greatly reduced, the higher temperature in this process tends to denature the protein, resulting in a decline in the quality of walnut protein.
The walnut fruit has not been pulverized by mechanochemical method, the specific surface area of ​​the particles is not uniform, and there is a problem of insufficient extraction
Supercritical CO 2 The extract was not subjected to selective adsorption of macroporous resin and gradient elution of different concentrations of ethanol, it may not be possible to obtain walnut protein with high purity and wide range of protein polarity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment example 1

[0027] 10g of walnut kernels were mechanically pretreated for 20 minutes and ball-milled into powder, degreased with petroleum ether, mixed with 60ml of 30% ethanol solution, stirred well, and added 0.005g of papain hydrolase at 45°C and pH=7.0 solution for 15 minutes to obtain the crude extract; use supercritical fluid carbon dioxide to extract the crude extract to obtain walnut protein extract, wherein the conditions of supercritical fluid extraction are: supercritical temperature is 45 ° C, supercritical pressure is 25MPa, extract Time is 0.5h, CO 2 The flow rate is 6L / min; use D3520 macroporous resin to adsorb the extract for 1 hour, and then use 5% ethanol, 15% ethanol, 20% ethanol, 25% ethanol and 35% ethanol to absorb the extract on D3520 macroporous resin. The filtrate is eluted, the eluents after different gradient elutions are collected, and the walnut protein eluate is obtained after merging; the walnut protein eluate is concentrated under reduced pressure, and drie...

Embodiment example 2

[0029] 10g of walnut kernels were mechanically pretreated for 20 minutes and ball-milled into powder, degreased with petroleum ether, mixed with 60ml of 20% ethanol solution, stirred well, and added 0.005g of papain hydrolase at 40°C and pH=6.5 solution for 30 minutes to obtain the crude extract; use supercritical fluid carbon dioxide to extract the crude extract to obtain walnut protein extract, wherein the conditions of supercritical fluid extraction are: supercritical temperature is 50 ° C, supercritical pressure is 25MPa, extract Time is 0.5h, CO 2 The flow rate is 6L / min; use D3520 macroporous resin to adsorb the extract for 1 hour, and then use 5% ethanol, 15% ethanol, 20% ethanol, 25% ethanol and 35% ethanol to absorb the extract on D3520 macroporous resin. The filtrate is eluted, the eluents after different gradient elutions are collected, and the walnut protein eluate is obtained after merging; the walnut protein eluate is concentrated under reduced pressure, and drie...

Embodiment example 3

[0031]10g of walnut kernels were mechanically pretreated for 30 minutes and ball-milled into powder, degreased with petroleum ether, mixed with 60ml of 40% ethanol solution, stirred well, and added 0.006g of papaya proteolytic enzyme under the conditions of 35°C and pH=6.0 Decompose for 20min to obtain the crude extract; use supercritical fluid carbon dioxide to extract the crude extract to obtain walnut protein extract, wherein the conditions of supercritical fluid extraction are: supercritical temperature is 55 ° C, supercritical pressure is 25MPa, extraction Time is 0.5h, CO 2 The flow rate is 6L / min; use D3520 macroporous resin to absorb the extract for 1 hour, and then use 5% ethanol, 15% ethanol, 20% ethanol, 25% ethanol and 35% ethanol to treat the filtrate adsorbed on the macroporous resin Carry out elution, collect the eluate after different gradient elutions, and obtain the walnut protein eluate after merging; carry out decompression concentration to the walnut prote...

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Abstract

The invention provides a method for quick separation and purification of walnut protein from walnuts. The method comprises the following steps that plant raw materials containing the walnut protein are subjected to ball milling and crushed into particles, thus walnut powder is obtained, and the dry weight is weighed; petroleum ether is adopted for degreasing, and an organic solvent is volatilized;an alcohol solution is adopted as an extracting solvent, and proteolytic enzyme is added for enzymolysis; the alcohol solvent is removed through rotary evaporation, then a supercritical fluid is adopted to extract the protein in the walnuts, and extracting liquid is adsorbed through macroporous resin; and alcohol solutions with the different concentrations are adopted to elute filtrate adsorbed on the macroporous resin, thus eluant is obtained, the eluant is combined, and vacuum concentration and freezing drying are conducted to obtain a walnut protein solid.

Description

technical field [0001] The invention relates to a method for rapidly separating and purifying walnut protein from walnuts. Background technique [0002] Walnuts (Juglans regia L), also known as walnut and Qiang peach, is a plant of the family Juglandaceae. Together with almonds, cashew nuts and hazelnuts, it is also known as the world-renowned "four major dried fruits". Walnuts are rich in nutrients, containing 15-20 grams of protein per 100 grams, more fat, 10 grams of carbohydrates, and contain beneficial calcium, phosphorus, iron and other trace elements and minerals, as well as carotene, Riboflavin, etc., is one of the nut foods loved by the common people. According to literature reports, walnut protein is nut protein, which is rich in resources and low in price, and is the main source of protein for Chinese residents. Among them, gluten accounts for 70%, globulin 18%, albumin 7% and other alcohol-soluble proteins. The proportion of 8 essential amino acids in walnut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/14
CPCC12P21/06C07K1/14Y02P20/54
Inventor 何建新王平龚俊杰王顺余郑宋友
Owner ZHEJIANG LIZIYUAN FOOD CO LTD
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