Method for preparing polypeptide by beer sediment

A technology of brewer's grains and wheat grains, which is applied in the field of preparing polypeptides by using brewer's grains, can solve the problems of preparation and activity research that have not been reported, and achieve the effect of obvious hypoglycemic activity and a wide range of sources

Active Publication Date: 2010-06-16
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are more and more researches on the biological activity of natural peptides, but there are no reports on the preparation and activity research of the rich source of brewer's grain peptides

Method used

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  • Method for preparing polypeptide by beer sediment
  • Method for preparing polypeptide by beer sediment
  • Method for preparing polypeptide by beer sediment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1 technological process is as figure 1 shown

[0030] (1) Vacuum freeze-dry the wet brewer's grains (provided by Guangzhou Zhujiang Brewery Group Co., Ltd.), pulverize them with a universal pulverizer, pass through a 100-mesh sieve, and obtain brewer's grains; Extracting agent, the extracting agent is 95% ethanol: 0.01mol / LNaOH=1: 2 (v: v), stirred at room temperature for 60min, filtered to remove residue, centrifuged at 2000rpm for 30min to obtain protein extract; The solution was adjusted to pH 4.5 with 0.2mol / L citric acid to carry out protein isoelectric precipitation, the supernatant was removed, and vacuum freeze-dried to obtain crude polypeptide;

[0031] (2) Add disodium hydrogen phosphate-citric acid buffer solution to the crude polypeptide obtained in step (1) to adjust the pH value to 6.5; add Alcalase alkaline protease (purchased from Novozymes), heat to 65 ° C, shake hydrolyzed under conditions for 1 h to obtain a hydrolyzate; the added amount ...

Embodiment 2

[0036] (1) Vacuum freeze-dry the wet brewer's grains (provided by Guangzhou Zhujiang Brewery Group Co., Ltd.), pulverize them with a universal grinder, pass through a 100-mesh sieve, and obtain brewer's grains; Extracting agent, the extracting agent is 70% ethanol: 0.10mol / LNaOH=1: 2 (v: v), stirred at room temperature for 120min, filtered to remove residue, centrifuged at 6000rpm for 10min to obtain protein extract; The solution was adjusted to pH 4.0 with 0.2mol / L citric acid to carry out protein isoelectric precipitation, the supernatant was removed, and vacuum freeze-dried to obtain crude polypeptide;

[0037] (2) Add disodium hydrogen phosphate-citric acid buffer solution to the crude polypeptide obtained in step (1) to adjust the pH value to 8.5; add Alcalase alkaline protease (purchased from Novozymes), heat to 45°C, and shake hydrolyzed under conditions for 2 hours to obtain a hydrolyzate; the added amount of the disodium hydrogen phosphate-citric acid buffer solution ...

Embodiment 3

[0042] (1) Vacuum freeze-dry the wet brewer's grains (provided by Guangzhou Zhujiang Brewery Group Co., Ltd.), superfinely pulverize them, and pass through a 100-mesh sieve to obtain brewer's grains; add 3000 mL of brewer's grains for extraction at a solid-to-liquid ratio of 1:30 per 100 g of brewer's grains. The extractant is 70% ethanol: 0.05mol / LNaOH=1:2 (v:v), stirred at room temperature for 70min, filtered to remove slag, centrifuged at 4000rpm for 20min to obtain protein extract; 0.2mol / L citric acid to adjust the pH to 5.0 for protein isoelectric precipitation, remove the supernatant, and vacuum freeze-dry to obtain the crude polypeptide;

[0043] (2) Add disodium hydrogen phosphate-citric acid buffer solution to the crude polypeptide obtained in step (1) to adjust the pH to 7.5; add Alcalase alkaline protease (purchased from Novozymes), heat to 50°C, and shake hydrolyzed under conditions for 5h to obtain a hydrolyzate; the added amount of the disodium hydrogen phosphat...

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Abstract

The invention discloses a method for preparing polypeptide by beer sediment. The method comprises the following steps: drying, pulverizing and sieving wet beer sediment to obtain beer sediment; adding extractant to the beer sediment and stirring the mixture at room temperature, filtering, deslagging and centrifuging the mixture to obtain protein extracting solution; adjusting pH value to 4.0-5.0 for precipitation; removing supernatant, carrying out vacuum freeze drying to obtain crude polypeptide; adding disodium hydrogen phosphate-citric acid buffer solution to the beer sediment crude protein, adjusting pH value of the mixture to 6.5-8.5; adding Alcalase alkaline protein enzyme, heating to 45-65 DEG C, carrying out hydrolysis for 1-5h while the mixture is stirred; afterwards killing enzyme in boiling water and centrifuging the mixture; taking centrifuged supernatant for gel column analysis and respectively collecting peaks and then carrying out concentration and freeze drying to obtain active polypeptide. In the method of the invention no harmful substance is introduced, the product is made of natural materials and enjoys wide material source, so that the natural active substances prove to have obvious blood-sugar-level-regulate activity according to in vitro experiments.

Description

technical field [0001] The method belongs to the field of food and health care, and particularly relates to a method for preparing polypeptides by using beer grains. Background technique [0002] Brewer's grains are one of the main wastes in beer brewing production. It is estimated that my country's beer production will reach 30 million tons / year in 2010, and the production of brewer's grains will also reach 7.5 million tons / year (wet grains). According to the measurement, its protein content is 23%~30% (dry basis), it can be seen that brewer's grain is a kind of good protein resource. For a long time, most manufacturers mainly sell the wet grains directly as roughage at a low price, with little profit; a few manufacturers directly discharge the brewer's grains, which not only causes serious environmental pollution, but also leads to a waste of resources. Therefore, the development and comprehensive utilization of brewer's grains has become an important task for researcher...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/14C07K2/00A61K38/02A61P3/10
CPCA61K38/02C12P21/06C07K2319/00A23J3/34A61K38/00C12F3/06A61P3/10
Inventor 李冰田应娟李琳陈玲朱良刘国琴苏健裕李晓玺
Owner SOUTH CHINA UNIV OF TECH
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