JPH089881A - Method for removing serum albumin in cow's milk - Google Patents

Abstract

PURPOSE:To effectively and simply remove bovine serum albumin from skimmilk and whey by bringing a resin containing an immobilized retinoid therein into contact with the bovine serum albumin. CONSTITUTION:This method for removing bovine serum albumin in cow's milk is to wash, e.g. aminated Cellulofine (R) with deionized water, replace the water with anhydrous ethanol, provide a suspension, add an anhydrous ethanol solution containing retinoic acid therein to the suspension, add dicyclohexylcarbodiimide as a condensing agent thereto, keep the pH of the mixed solution at 5-6, carry out the reaction while shaking the mixture at ambient temperature, prepare a resin containing the immobilized retinoid therein and remove the bovine serum albumin in the cow's milk utilizing the affinity of the bovine serum albumin for the retinoid.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a simple method for removing bovine serum albumin from skim milk and whey.

[0002]

[Prior Art] Since 1984, Borch-Johnsen et al.
hnsen, K. et al , Lancet, 2 , 1083-1086, 1984: Relat
ion between brest-feeding and incidence of insulin
-dependent diabetes mellitus.) found that there is a negative correlation between insulin-dependent diabetes mellitus and the period of breastfeeding in Norway and Sweden.

About this diabetes Karjalainen in 1992
(Karjalainen, J. et al , NewEngl. J. Med., 327 , 3
02-307,1992: A bovine albumin peptide as a possibl
etrigger of insulin-dependent diabetes mellitus)
On the primary structure of bovine serum albumin contained in milk
It was found that a peptide composed of 17 amino acids at the 152-168th position was absorbed by an infant, and an antibody against this peptide acts on β-cells that make insulin in the pancreas and destroys the cells. In this regard, they produce milk or modified milk made from it as a main ingredient
Warns that it should not be given to infants for a period of one to two years.

[0004]

However, there are not enough foods other than these for infants at present, and there is an urgent need to establish a method for removing bovine serum albumin from milk.

[0005]

[Means for Solving the Problems] Therefore, the inventors have been energetically conducting research on low allergen milk.
Among them, bovine serum albumin can be removed very easily by contacting retinoid-immobilized resin created for the purpose of removing β-lactoglobulin or resin having long-chain fatty acid immobilized with bovine serum albumin. Headline,
Completed this invention.

[0006]

In the method for removing bovine serum albumin from milk according to the first aspect, the affinity of bovine serum albumin with retinoid by the resin on which retinoid is immobilized is utilized.
In the method for removing bovine serum albumin from milk according to the second aspect, a resin in which a long chain fatty acid such as linoleic acid or linolenic acid is immobilized is used and its affinity is utilized. In the method for removing bovine serum albumin from milk according to claim 3, skim milk is contacted with the resin according to claim 1 or 2. In the method for removing bovine serum albumin from milk according to claim 4, whey is brought into contact with the resin according to claim 1 or 2.

[0007]

【Example】

Example 1) Preparation of retinoic acid-immobilized resin: 10 ml of aminated cellulophane was thoroughly washed with deionized water and replaced with anhydrous ethanol to obtain a suspension of 10 ml. Anhydrous ethanol solution containing retinoic acid at a concentration of 1 mg / ml was added, and 0.5 g of dicyclohexylcarbodiimide was added as a condensing agent.
The pH of the mixed solution was maintained at 5 to 6, and the reaction was carried out for 16 hours at room temperature with gentle shaking. After completion of the immobilization reaction, the resin was washed with anhydrous ethanol and suspended in deionized water,
It was washed with 0.1N hydrochloric acid. It was replaced with deionized water, and this was used as a retinoic acid-immobilized resin.

Next, retinoic acid-immobilized resin (1 g) was suspended in skim milk (25 ml), and the retinoic acid-immobilized resin was filtered off on a glass filter or filter paper. As shown in Table 1, the amount of bovine serum albumin in the filtrate obtained at this time was SDS.
-Electrophoresis, Western blotting and ELISA
It was not detected by the method.

[0009]

[Table 1]

Example 2) Preparation of linoleic acid-immobilized resin: 10 ml
The aminated cellulophane of was thoroughly washed with deionized water and replaced with anhydrous ethanol to give a suspension of 10 ml. 1m
Add an ethanol solution containing linoleic acid at a concentration of g / ml,
0.5 g of dicyclohexylcarbodiimide was added as a condensing agent. The pH of the mixed solution was maintained at 5 to 6, and the reaction was carried out for 16 hours at room temperature with gentle shaking. After the completion of the immobilization reaction, the resin was washed with anhydrous ethanol, suspended in deionized water, and then washed with 0.1N hydrochloric acid. Replace with deionized water,
This was designated as linoleic acid-immobilized resin.

The contents of bovine serum albumin and β-lactoglobulin in the skim milk (25 ml) used in Example 1 and the skim milk treated with the linoleic acid-immobilized resin immobilized in the same manner as in Example 1 are shown in Table 2. As shown in (3), it was not detected by SDS-electrophoresis, Western blotting and ELISA.

[0012]

[Table 2]

Example 3) A whey was obtained by an isoelectric point or the action of a rennet according to a conventional method, and the pH of whey (50 ml) was adjusted to 6.8 with 0.1N sodium hydroxide, and then retinoic acid-immobilized resin.
(1 g) was added and suspended. The bovine serum albumin and β-lactoglobulin contents of this retinoic acid-treated whey were not detected by any of the methods as shown in Table 3.

[0014]

[Table 3]

Example 4) The whey (50 ml) used in Example 3 was brought into contact with the linoleic acid-immobilized resin (1 g), and the linoleic acid-immobilized resin-treated whey was treated by the method described in Example 1. Obtained. The bovine serum albumin and β-lactoglobulin contents in the whey obtained at this time were not detected by any of the methods as shown in Table 4.

[0016]

[Table 4]

[0017]

According to the above method of the present invention, bovine serum albumin in milk can be removed very effectively.

Claims (4)

[Claims] 1. A method for removing bovine serum albumin from milk utilizing the affinity of bovine serum albumin for retinoid with a resin on which retinoid is immobilized. 2. A method for removing bovine serum albumin from milk by using a resin in which a long chain fatty acid such as linoleic acid or linolenic acid is immobilized and utilizing its affinity. 3. A method for removing bovine serum albumin from milk by bringing skim milk into contact with the resin according to claim 1 or 2. 4. A method for removing bovine serum albumin from milk by bringing whey into contact with the resin according to claim 1 or 2.