JP2000106826A - Composition containing chlorella peptide - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a composition containing chlorella peptide, having biological activity, high digestive absorption ratio and excellent taste, flavor and nutrition and useful for health food, etc., by treating chlorella with a proteolytic enzyme, separating the decomposition product into solid and liquid and recovering the liquid part. SOLUTION: Chlorella such as algae of the genus Chlorella is treated with a proteolytic enzyme originated e.g. from microorganism of the genus Aspergillus, the obtained enzymatic decomposition product is separated into solid and liquid and the liquid part is recovered and preferably concentrated and/or dried to obtain the objective chlorella-containing composition having the following properties. Peptide content, 65-95%; molecular weight fraction, peptide having a molecular weight of <=10.000 and composed mainly of a peptide having a molecular weight of 955 and a peptide having a molecular weight of 279 and water-soluble nucleic acid; digestivity (compared with starting chlorella), the digestivity with pepsin is almost 100%; physiological activity, positive ACE inhibiting activity; bitter taste, scarce; solubility, soluble in wter. Preferably, a health food or drink is produced by using the composition as a component.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to the production of a chlorella peptide-containing composition. According to the present invention, since the active ingredient in chlorella cells is solubilized and released outside the cells, completely different from conventional chlorella products,
In addition to flavor and nutrition, it is possible to produce new compositions that are excellent in terms of physiological activity and digestion and absorption, and can be used not only for ordinary foods and drinks, seasonings, but also for various foods and drinks such as health foods and nutritional foods. It can also be used for applications.

[0002]

2. Description of the Related Art Many chlorella products are commercialized with one cell surrounded by a cell membrane and a cell wall because chlorella is a kind of algae, and is extremely excellent in nutritional aspect. However, it was difficult to digest and absorb in vivo. Chlorella in which the cell wall has been crushed has been developed in order to increase the digestibility and absorbability in the living body. However, this also has a cell membrane remaining, and is not yet sufficient for the digestibility and absorbability in the living body. Furthermore, chlorella products having a high peptide content and containing a water-soluble nucleic acid have not been known at all.

[0003]

SUMMARY OF THE INVENTION In view of the above-mentioned state of the art, the present invention has been made for the purpose of developing a novel chlorella product which has not been known so far, and to further effectively utilize chlorella which is originally known as a useful natural resource. Things.

[0004]

Means for Solving the Problems In order to achieve the above object, the present inventors have studied from various aspects. As a result, when the chlorella raw material was treated with a protease, a double barrier consisting of a cell membrane and a cell wall of chlorella cells was obtained. And obtained a very useful knowledge that the active ingredient in the cell is released after passing through.

Furthermore, the active ingredient is not only water-soluble, but also has an excellent property that it has a high peptide content, contains nucleic acid, has no bitterness, and also has an excellent ACE inhibitory activity. Were obtained, and further studies were conducted based on these useful new findings, and the present invention was completed. Hereinafter, the present invention will be described in detail.

[0006] Chlorella raw materials include Chlorella algae (for example, Ch.
lorella pyrenoidosa, C. ellipsoidea, C. vulgaris
) Can be used, and chlorella raw materials commercially available not only from Japan but also from foreign countries such as from Taiwan can be used freely. It is also preferable to use raw materials in powder form.

In order to carry out the present invention, it is necessary to treat a chlorella raw material with a protease. Specifically, when a powdered chlorella raw material is used, after adding 10 to 30 times the amount of water, an enzymatic decomposition reaction may be performed under suitable reaction conditions according to the enzyme to be used.

[0008] As the enzyme, any enzyme can be used as long as it is a proteolytic enzyme used in the food manufacturing industry, such as those derived from microorganisms and plants, and non-limiting examples include the following: Enzymes derived from Aspergillus (Protease P-6, Sumizyme LP, Flavorzyme, Denazyme AP, Punchidase: all trade names), Enzymes derived from Bacillus (Esperase, Protin PC-10, Neutrase, Bioprase: all) Product name).

[0009] In addition to the above, all enzymes capable of degrading proteins can be used alone or in combination as the protease. Its origin can be obtained from microorganisms in addition to animals and plants, and in addition to pepsin, renin, trypsin, chymotrypsin, papain, bromelain, bacterial proteases, filamentous fungal proteases, actinomycetes proteases and the like can be widely used. As these enzymes, those commercially available are generally used, but unpurified enzymes, culture solutions containing the enzymes, solid or liquid enzyme contents such as koji may also be used as necessary according to the purpose. Can be.

[0010] The amount of the proteolytic enzyme varies depending on the degree of purification of the enzyme and also varies depending on the treatment time, but it is sufficient to use 0.05 to 5%, preferably about 2% of the treatment solution. Further, as will be apparent from the following description, an enzyme derived from a microbial system, in particular, a microorganism of the genus Aspergillus is effective for obtaining a composition having less bitterness, and is suitable as the enzyme used in the present invention. After the enzyme treatment, heat is applied to inactivate the enzyme.

The treatment liquid for stopping the enzyme reaction is filtered through a vibro screen or the like, and if necessary, subjected to a Jector treatment.
Using a Sharpless centrifuge or the like, for example, 10,000
Centrifuge at ３０30,000 rpm and microfilter if necessary.

This is concentrated by vacuum concentration or the like (1 to 30 Bx
) To obtain a chlorella peptide stock solution, if necessary. The chlorella peptide stock solution can be used for various applications as it is. However, if necessary, the chlorella peptide stock solution can be purified by activated carbon filtration, diatomaceous earth filtration or the like, and used as it is as a chlorella peptide stock solution. The peptide stock solution obtained here can be further purified with alcohols, ion-exchange resin, etc., but these purifications are not particularly required, and are purified to the extent that they can be used as active ingredients for various applications. Is what it is.

The chlorella peptide stock solution may be further sterilized and filled, if necessary, and then dried and solidified according to a conventional method. For example, chlorella peptide powder can be obtained by freeze-drying or spray-drying with or without the addition of dextrin to obtain a chlorella peptide powder.

Thus, a chlorella peptide-containing composition such as a liquid chlorella peptide-containing composition (chlorella peptide stock solution) and a solid chlorella peptide-containing composition (chlorella peptide powder) can be obtained.
FIG. 1 shows an example of a method for producing the present composition.

[0015] The present composition contains an active ingredient in chlorella cells, the protein is decomposed into peptides, the content of water-soluble nucleic acid is greatly increased, and other components are all different from conventional chlorella products. In addition to being a water-soluble component, it has no bitterness, has an extremely high digestibility, and can be used as it is or as a food or drink when mixed with other components. As described above, the present composition has a remarkable characteristic of a water-soluble chlorella product, so that it can be used as a beverage, and is excellent in this respect.

Further, since the present composition contains a peptide and a water-soluble nucleic acid, it has a taste and can be used as a seasoning. Furthermore, the water-soluble nucleic acid is CG
It is also referred to as F (Chlorella Growth Factor), and is expected to have a physiologically activating effect such as a cell activating effect and an immunostimulating effect. In addition, the present composition contains a chlorella peptide, and further contains an angiotensin I converting enzyme ( ACE) also has an effect of suppressing the activity of ACE), and therefore has a blood pressure lowering effect. Therefore, the present composition can be used not only as ordinary food and drink, but also can be effectively used as health food and drink and health food and drink.

At this time, the composition uses chlorella used as food as a raw material, and does not use any substances that are a problem in food safety in the process of processing. In addition, not only ordinary foods and drinks but also health foods and drinks can be used safely by elderly people and the sick. Hereinafter, examples of the present invention will be described.

[0018]

Example 1 A commercial product made in Taiwan was used as a raw material of chlorella, which was degraded with a commercially available protease shown in Table 1 below, and a degradation test of chlorella was performed.

[0019]

[Table 1]

In the decomposition test of chlorella, a 10-fold amount of water was added to the chlorella raw material according to the decomposition conditions shown in FIG.
Decomposed with the above-mentioned commercially available protease,
After heating for one minute, the supernatant was separated by screen treatment, and the supernatant was measured for protein yield, peptide content, and the like. Table 2 shows the obtained results. As a blank, a sample obtained by boiling and extracting a 10-fold amount of hydrochloride of the chlorella raw material for 10 minutes was also processed and measured in the same manner as in the case of the enzymatic decomposition described above.

[0021]

[Table 2]

For the above five enzymes having relatively high protein yields or low bitterness, Aspergillus genus such as punchase, protease P-6, and flavorzyme are evident from the results in Table 2 in which the peptide contents were measured. The enzymes of origin have been found to be particularly suitable for the degradation of chlorella. Among them, protease P-6 had the best taste and decomposition power.

Next, high-performance liquid chromatography (HPLC) was carried out on the “boil extract” of the raw material chlorella obtained above and “chlorella peptide” obtained using protease P-6 under the following analysis conditions. The components were compared. FIG. 3 shows the obtained results.

(Analytical conditions) Measurement: HPLC Pump: Shimadzu LC-9A Detector: Shimadzu SPD-6AV Column: Asahipack GS-320 (15 cm × 2 connected + 5 cm) Mobile phase: 0.1% TFA-45% acetonitrile aqueous solution Flow rate: 0.5 ml / min Detection wavelength: 260 nm

As shown in FIG. 3 (Molecular weight fraction of Chlorella), the data of high performance liquid chromatography showed that “Chlorella peptide” indicated by a dotted line was compared with “Boiled extract” indicated by a solid line.
However, it can be seen that both the peptide component and the nucleic acid component are contained in large amounts. "Boiled extract" contains a protein with a molecular weight of about 60,000. Enzymatically degraded "Chlorella peptide" has almost no protein, and it is a soluble peptide fraction (molecular weights 955 and 279). I understand. That is, the molecular weight was mostly 10,000 or less, and the di- and tripeptides were the main.

From the molecular weight fractionation data, the amounts of peptides and nucleic acids were calculated according to the following equations. Peptide amount = peptide peak area of "chlorella peptide" / peptide (protein) peak area of boiled product = 380506/104834 = 3.63 times Nucleic acid amount = nucleic acid peak area of "chlorella peptide" / nucleic acid peak area of boiled product = 239370/31662 = 7.56 times As is clear from the above, the peptide content is about 3.6 times that of the “boiled product” and the nucleic acid content (CGF) is about 7 times that of the “boiled product” by the protease treatment. .6 times.

[0027]

Example 2 A 10-fold amount of water was added to a chlorella raw material (produced in Taiwan), 2% of protease P-6 (commercial name of a commercially available protease agent) was added, and the mixture was kept at 50 ° C. for 3 hours to perform enzymatic degradation. Was. The mixture was then boiled for 15 minutes to inactivate the enzyme. This is filtered with a vibro screen (150 mesh),
The filtrate was subjected to a Jector treatment at 5,000 rpm, followed by treatment with a Sharpless centrifuge (15,000 rpm).
Microfiltration using a press filter using diatomaceous earth as a filter aid, concentration under reduced pressure to Bx 7.0, and liquid chlorella peptide-containing composition (chlorella peptide stock solution)
I got

Subsequently, this was sterilized on a plate at 120 ° C. for 5 seconds, spray-dried, and dried to obtain a chlorella peptide-containing composition (chlorella peptide powder).
I got

The component analysis values of the chlorella raw material are shown in Table 3 below, and the component analysis values of the chlorella peptide powder prepared as described above are shown in Table 4 below. Among the protein contents of the chlorella raw material, those obtained as "solubilizing components" are 21% in the case of boiling extraction (blank),
In the case of the enzyme treatment, it was 40 to 62% (the measured data width of various enzymes), and particularly in the case of protease P-6, it was 57%.

[0030]

[Table 3]

[0031]

[Table 4]

As is evident from the above results, the “chlorella peptide” obtained by enzymatic degradation with “protease P-6”
As shown in the component analysis values of No. 4, the raw material chlorella contained 4.6% of "water-soluble nucleic acid" which was little. This “water-soluble nucleic acid” is called CGF (Chlorella Growth Factor) and is one of the active ingredients.
It is said that it has physiological activities such as "cell activation". The other components are not significantly different from the raw material chlorella, and are all water-soluble components. Therefore, it can be said that only the components that can be digested and absorbed among the intracellular components of the raw material chlorella are extracted.

[0033]

Example 3 For the purpose of proving that the human digestibility of chlorella peptide was superior to that of chlorella raw material, the digestibility of chlorella peptide pepsin was measured as follows.

A pepsin-HCl solution (0.075N pepsin protein ratio of 0.6) was added to a sample (chlorella peptide (chlorella peptide stock solution prepared in Example 2) and chlorella raw material).
%), And the total amount was adjusted to 6 ml. After centrifugation, the amount of protein in 1 ml of the supernatant was measured by the Lowry method. Then 45 ° C
After incubation for 4 hours, the mixture was centrifuged again at 3000 rpm, and the amount of protein in 1 ml of the supernatant was measured. The digestibility was calculated by the following equation. FIG. 4 shows the obtained results. Digestibility (%) = (Amount of protein in 1 ml of centrifuged supernatant / amount of protein in 1 ml of sample lysate) × 100

As is clear from the results of FIG. 4 (digestibility of pepsin), chlorella is usually solubilized by pepsin. Is considered to be less than half the intake. On the other hand, since “chlorella peptide” produces only soluble components, it is 100% solubilized from the beginning and has conditions for 100% digestion and absorption. That is, it can be seen that “chlorella peptide” has a higher digestibility (not decomposed by pepsin) as compared with the chlorella raw material.

"Chlorella peptide" not only has a high digestibility as described above, but also has a high ACE inhibitory activity, and is useful as a hypotensive composition. As a result of measuring the ACE inhibitory activity according to the following method, 4.30 mg / m
l. On the other hand, SP-100 is 1.05m
g / ml, 2.76 mg / ml of casein enzyme degradation product
Met. ACE inhibitory activity measurement method: ACE (manufactured by Sigma, derived from rabbit lung) 2.5 mU, Synthetic substrate Hippuryl, Institute for Protein Research
Matsui et al. (T. Matsui, H. Matsufuji and Y. Osajima: Biosc) using 12.5 mM L-histidyl-L-leucine.
i. Biotech. Biochem., 56, 517 (1992)). The ACE inhibitory activity IC ₅₀ value of the sample was represented by the protein concentration of the sample required to inhibit ACE enzyme activity by 50% (inhibition rate).

[0037]

Example 4 A tablet-type chlorella peptide-containing composition product was produced using the following raw materials. (1) Chlorella peptide powder (Example 2) 10 g (2) Egg yolk powder 3 g (3) Lactose 87 g (4) Corn starch 29 g (5) Magnesium stearate 1 g (1), (2), (3) and 17 g of corn starch Was mixed and granulated together with a paste made from 7 g of corn starch, 5 g of corn starch and (5) were added to the granules, and the resulting mixture was tableted with a compression tablet machine to produce 100 tablets.

[0038]

Example 5 Chlorella peptide stock solution (Example 2) 20k
g, egg yolk 30 kg, and granulated sugar 10 kg, and the total amount was made 100 kg. Then 90 ~
Sterilize at 95 ° C. for 15 to 30 seconds, rapidly cool the product temperature to about 70 ° C., fill 100 g of each in a pre-sterilized brown bottle, immediately seal the bottle, and 1000 drink-type chlorella peptide-containing composition products. Manufactured.

[0039]

According to the present invention, the following remarkable effects are obtained. The active ingredients in the cells are released outside the cells by enzymatically decomposing the chlorella raw material with proteolytic enzymes.
By concentrating or drying only the active ingredient, lossless ingestion becomes possible. Since only the intracellular active ingredient can be commercialized, the product has a very high digestibility and is highly absorbable.
The range of application as chlorella is increased. For example, the present invention can be applied to beverages which are conventionally insoluble and thus difficult to use.

[Brief description of the drawings]

FIG. 1 shows a method for producing chlorella peptide.

FIG. 2 shows degradation conditions of chlorella with a protease.

FIG. 3 is a molecular weight fractionation diagram of Chlorella.

FIG. 4 shows the pepsin digestibility of Chlorella.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A23L 2/52 A23L 2/38 F 2/66 2/00 F 2/38 J

Claims (7)

[Claims] 1. A method for producing a chlorella peptide-containing composition, comprising: treating chlorella with a protease; performing solid-liquid separation on the obtained enzyme degradation product; and separating and collecting a liquid portion. 2. The method according to claim 1, wherein an enzyme derived from a genus Aspergillus is used as the protease. 3. The method according to claim 1, wherein the liquid part is further concentrated and / or dried. 4. A chlorella peptide-containing composition produced by the method according to any one of claims 1 to 3. 5. A chlorella peptide-containing composition comprising a liquid part of an enzymatically degraded product obtained by treating chlorella with a proteinase derived from a genus Aspergillus or a processed product thereof and having the following properties. (A) Peptide content 65 to 95% (b) Molecular weight fraction Peptide having a molecular weight of 10,000 or less, centered on a peptide having a molecular weight of 955, a peptide having a molecular weight of 279, and a water-soluble nucleic acid (c) Digestibility (with the raw material chlorella) Comparison) Digestibility by pepsin: almost 100% (d) Physiological activity ACE inhibitory activity: yes Bitter taste: almost none (e) solubility water solubility 6. A food or drink comprising the chlorella peptide-containing composition according to claim 4 or 5. 7. The food or drink according to claim 6, wherein the food or drink is a health food or drink.