JPH0445750A - Production of peptide-containing article for food - Google Patents

Abstract

PURPOSE:To obtain the title article having excellent deodorizing property and decolorizing property and useful as a food raw material, etc., by treating a crude peptide-containing material for food with a silica gel to which lipophilic group is bound. CONSTITUTION:A crude peptide-containing material (e.g. sardine meat) for food is treated with a silica gel (preferably having 100-500mum grain size and 100-300Angstrom pore size) to which a lipophilic group such as octadecyl group is bound and deodorized and/or decolorized to provide the aimed article.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a method for obtaining peptide-containing articles that are easy to use as foods by removing undesirable colors and/or odors derived from raw materials.

[Conventional technology]

Vep comes from food. Tides are diverse, with different molecular weights (hundreds to thousands), physicochemical properties, and physiological functions depending on their origins and purification methods.

In recent years, the effects of food ingredients on living organisms, i.e.
The concept of evaluation based on ``functionality'' has been formed, and peptides as food ingredients are also attracting attention. Food functions related to peptides include the primary function of small peptides, which is superior intestinal absorption ability compared to amino acids, secondary functions such as taste and food properties, promotion of calcium absorption, immune enhancement, lipid metabolism improvement, and blood pressure. There are tertiary functions of bioactive peptides such as descent.

When peptides in foods or food raw materials are separated and purified using current food manufacturing technology, the odor and color of the raw materials remain, and the separation accuracy is low, causing a decrease in product value. These deodorizing and decolorizing techniques include adsorption methods using activated carbon, extraction methods using organic solvents, and methods that decompose causative substances using microorganisms or enzymes.
Many problems remain.

[Problem to be solved by the invention]

Among the above-mentioned conventional techniques, the adsorption power of activated carbon varies greatly depending on the size of its pores, but no satisfactory deodorization or decolorization of peptide-containing materials has been achieved. In addition, other methods mentioned above are expensive for food processing and difficult to process in large quantities, and an effective alternative method has been desired.

The purpose of the present invention is to highly deodorize and decolorize peptide-containing substances, which have conventionally been limited in their use in food products due to their odor and color, so that they can be used in many foods.
The objective is to provide food materials with high added value by further purifying physiologically active peptide-containing materials and the like.

[Means to solve the problem]

To summarize the present invention, the present invention relates to the production of a deodorized and/or decolorized food-grade peptide-containing article by treating a crude food-grade peptide-containing material with silica gel bonded with a lipophilic group. Regarding the method.

As a result of extensive research to solve the above problems, the present inventors succeeded in fractionating deodorized and decolorized peptides by treating them with silica gel bonded with lipophilic groups. The invention has been completed.

The silica gel bonded with a lipophilic group of the present invention is one in which a lipophilic group is chemically bonded to the silanol group of silica gel, and the lipophilic group contains an octadecyl group (C, or 0D
S), octyl group (C6) butyl group (C,)
) Limethylsilyl group (TMS) Phenyl group (Ph
) There are cyanopropyl groups (CN), aminopropyl groups (Nli2), etc., and each has different lipophilicity and can be used depending on the target separation component. Among them,
The most versatile group is octadecyl. The shape of the silica gel is not particularly limited, but it is preferably particulate, and more preferably has a particle diameter of 100 to 500 μm and a fine diameter of 100 to 300 μm.

A specific method for using the lipophilic group-bonded silica gel (granules) is to thoroughly wash it with a hydrophilic organic solvent or an aqueous solution containing the same, and then thoroughly replace it with deionized water. Add it to the crude peptide-containing solution, stir well, and then filter it, or pass the crude peptide-containing solution through a column filled with it to remove the lipophilic components (odorous components). components, colored components, lipophilic peptides). Finally, the silica gel is washed with deionized water to wash away the remaining non-adsorbed components to obtain a deodorized and/or decolored hydrophilic food-grade peptide-containing article. The adsorbed peptide can be fractionated by elution with an aqueous solution in which the concentration of a hydrophilic organic solvent is gradually increased.

The origin of the crude peptide content in the present invention is animal,
Either plants or microorganisms may be used, but it is preferable to use those in a solution state. The solvent may be water or a hydrophilic solvent (such as ethanol) that can be used in foods.

The peptide-containing article for food according to the present invention can be used for general foods, but is preferably used for beverages, health foods, etc., and is particularly suitable for obtaining useful substances such as angiotensin enzyme (hereinafter abbreviated as ACE) inhibitors. It is suitable for use in beverages, health foods, etc.

The method of analyzing the product in the present invention is as follows.

(1) Peptide amount Total nitrogen was measured by the Kerbourg method, and the value obtained by multiplying it by 6.25 was used.

(2) Average number of amino acid residues (1) The average number of amino acid residues (1) was determined by the following formula.

1 = (Number of moles of free amine groups in the hydrochloric acid decomposition solution of sample 1)/(Number of moles of free amino groups in sample 1) Hydrolysis was carried out with 6N hydrochloric acid at 10°C for 24 hours to remove amino groups. Quantification was performed by the TNBS method.

(3) Put 50 μβ of ACE inhibitory activity sample in a test tube, add 100 μβ of ACE
E (manufactured by Sigma, 2.5 mM) solution was added, and 37
After incubation for 5 hours at °C, 100 μf of
ly-L-His-L-Leu (Peptide Research Shinsei,
A final concentration of 5 mJ NaCl (containing 400 mM) was added, and the mixture was reacted at 37°C for 60 minutes. Thereafter, 0.25-mL of 0.5N hydrochloric acid was added to stop the reaction, and then 1.5 tons of ethyl acetate was added and vigorously stirred for 15 seconds. Thereafter, it was centrifuged at 3000 rpm for 10 minutes, and the ethyl acetate layer was heated at 140° C. for 20 minutes to remove the solvent. After removing the solvent, it was dissolved in 3-IM NaCl aqueous solution, and the absorption of the extracted hippuric acid (absorbance of 228no+) was measured, and this was taken as the enzyme activity.

Inhibition rate = (A-B) /Ax 100 (%) A; Absorbance at 228 nm when no inhibitor is included B; Absorbance at 228 nm when an inhibitor is added And the inhibitory concentration when the inhibition rate is 50% is IDs . Closed.

〔Example〕

Hereinafter, the present invention will be explained in more detail with reference to Examples.
The present invention is not limited thereto.

Example 1 Water IIl was added to 100 g of ground sardine meat, 2 g of Denazym AP manufactured by Nagashio Sangyo ■ was added, and enzymatic decomposition was performed at 40° C. for 5 hours. Next, the enzyme was inactivated by boiling at 95°C for 15 minutes, cooled, centrifuged to degrease, and filtered through Celite to obtain a filtrate of 1.050W11. I got it. This filtrate is ODS
Silica gel [■YMC Gel 0DS-A manufactured by YMC
After passing the solution through a column filled with 125 g of Q 120-550, it was washed with 250 g of deionized water. This cleaning solution was added to the above-mentioned solution to prepare a sensory test sample.

As a control, the above filtrate was treated in the same way with activated carbon for deodorization and decolorization (manufactured by Takeda Pharmaceutical Co., Ltd., Shirasagi Y), which is commonly used for food products, and the resulting sample and the previous sample were evaluated in a sensory test. The degree of deodorization and decolorization was compared. The sample was a 5% aqueous solution, the panel consisted of 8 people, and the test method was a 3-point method. The average values of the results are shown in Table 1.

Table 1 (1; Good 2; Fair 3; Poor) Example 2 After passing the enzymatically decomposed solution of sardine surimi through an ODS silica gel column in the same manner as in Example 1, the passing solution was mixed with deionized water (in the column name). (20 times volume) washing solution was combined to obtain fraction 1. Next, the solution eluted with 10% acetonitrile (same) was fractionated into fraction 2.
The solution eluted with 25% acetonitrile (same) was divided into fraction 3.5.
The solution eluted with 0% acetonitrile (same) was fraction 4.99.
% acetonitrile (same) as fraction 5.

Color of these fractions (absorbance at 430r+m)
ACE inhibitory activity (ID,.) Peptide yield (N yield),
Table 2 shows the results of the study on the average number of amino acid residues and sensory tests.

As a result, the target peptide-containing substance was contained in fraction 1, which was well deodorized and decolored compared to the original solution.

Furthermore, of the two fractions adsorbed on the column, fraction 2, which had a stronger ACE inhibitory activity, was also deodorized and decolored compared to the original solution.

Example 3 Commercially available yeast extract [manufactured by Narita Pharmaceutical Co., Ltd., 5L-W] 20
Dissolve g in water 100%, ODS silica gel [manufactured by Fuji Gibson ■, Chromatorex-〇DS DM102
It was applied to a column packed with 15 g of 0T, and after passing through the column, it was washed with 50 d of deionized water. Thereafter, in the same manner as in Example 1, powders from the test group and the control group were obtained and subjected to a sensory test, and the test group was clearly deodorized and decolored compared to the control group.

Example 4 Fish sauce batis from the Philippines (manufactured by Unifish Export and Inboat Corporation) 100
In the solution obtained by treating 1 nl in the same manner as in Example 3, the test area was clearly deodorized and decolored compared to the control area.

Example 5 In the extract obtained by treating the aqueous solution 100-, which is a 5-fold dilution of meat extract and -fu (manufactured by Kyokuto Seiyaku Aji) in the same manner as in Example 3, the test group showed a clear difference compared to the control group. scent,
The colors have also been improved.

Example 6 20g of crushed whole soybeans and 200IL of water! and 200 μl of a commercially available proteolytic enzyme were added and hydrolyzed at 40° C. for 3 hours. After decomposition, the enzyme was deactivated by boiling at 95° C. for 15 minutes, and then cooled. After centrifuging the liquid, a clear liquid was obtained by celite filtration, and the same silica gel 10
After passing through the column, add 100 g of deionized water.
Washed with. This cleaning solution was added to the flow through to form a sensory test sample. The results are shown in Table 3.

Table 3 (10+; strongly perceived soybean odor, -8 not detected) This shows that the ODS treated solution was very well deodorized and decolored compared to the stock solution.

Example 7 Next, the deodorizing and decolorizing effects of various chemically bonded silica gels were examined and compared.

A column was filled with 2 g of each of the following resins, thoroughly washed with an aqueous ethanol solution (approximately 70%), and then thoroughly replaced with deionized water.

■TMS bond type; ChromaDrex TMSMO306 ■C1 bond type; same TET DM1020T ■C6 bond type; same OCT DM1020T ■C1° bond type; same O
DS DM1020T sample is sardine peptide stock solution (
peptide content of 1.6%), 40-mL of the column was applied to each column and washed with 20-mL of water. The cleaning solution was used as a sample along with the passing solution, and the chromaticity and sensory tests were compared and examined. The result is the fourth
Shown in the table.

Table 4 (+++ H Strong fishy odor, ++; felt, --not felt) As is clear from Table 4, Cla%C@, C4,7MS
The effects of deodorization and decolorization were greatest in the order of treatment.

〔Effect of the invention〕

As described above, by using the technology of the present invention, it becomes possible to efficiently deodorize and decolorize various crude food-grade peptide-containing products, and the versatility of these food products is expanded. The present invention is a very useful technique.

Claims (1)

[Claims] 1. A method for producing a deodorized and/or decolorized food-grade peptide-containing article, which comprises treating a crude food-grade peptide-containing substance with silica gel bonded with a lipophilic group.