JPH05284937A - Digestive enzyme activity-inhibiting substance extracted from sea alga and dietary food containing the same - Google Patents

Abstract

PURPOSE:To provide the subject inhibiting substance inhibiting the activities of alpha-amylase and lipase and effectively inhibiting obesity and hyperglycemia as a dietary food by filtering the extraction solution of a sea alga and subsequently purifying the filtrate. CONSTITUTION:A brown alga (e.g. Undaria pinnatifida, sea tangle), a green alga (e.g. Ulva pertusa, Enteromorpha) or a red alga (e.g. Gracilaria, Gelidium subcostatum) is hot-extracted with a solvent, preferably water or an alcohol, and subsequently filtered. The filtrate is purified to provide the objective inhibiting substance.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a digestive enzyme (α-amylase, lipase etc.) activity inhibitory substance extracted from seaweed and a diet food containing the digestive enzyme activity inhibitory substance.

[0002]

BACKGROUND OF THE INVENTION α-Amylase is an enzyme that hydrolyzes α-1,4-glucoside bonds such as starch and glycogen. It is widely present in various organisms and saliva in the human body. There are α-amylase derived from the pancreas and α-amylase derived from the pancreas, which play a role of converting starch into sugar in the oral cavity or the digestive tract, respectively. The α-amylase inhibitor inhibits the activity of this α-amylase, and therefore is an antiobesity diet drug,
It is useful as a therapeutic agent for hyperglycemia, a therapeutic agent for diabetes, a preventive agent for dental caries and the like. For this reason, it has been attempted to extract and obtain α-amylase inhibitors from various raw materials, and for example, a method for extracting α-amylase inhibitors contained in wheat is known (Japanese Patent Application Laid-Open No. 2004-242242). JP-A-57-140727, JP-A-60-41
No. 32, etc.). On the other hand, lipase is an enzyme that hydrolyzes fat and is present in pancreatic juice in the human body. Lipase inhibitors block the activity of this lipase,
It is useful as an anti-obesity diet agent, a therapeutic agent for hyperlipidemia and the like.

As mentioned above, α-in wheat or flour.
It is known that an amylase inhibitor is contained, but in order to remove this α-amylase inhibitor from wheat or flour, a water extract of wheat or flour is heat-treated, fractionated by an organic solvent, and precipitated. The desired α-amylase inhibitor is obtained by a very complicated operation of collecting a portion, treating the solution with an adsorbent, eluting with a salt solution, and fractionating by chromatography. Not a method.
Further, the α-amylase inhibitor obtained by such a method is inevitably expensive, and is therefore unsuitable for use as a dieting agent which requires providing a large amount of the α-amylase inhibitor at low cost. there were. In addition, the conventionally obtained α-amylase inhibitor is
Whether or not it has an effect of inhibiting the activity of not only α-amylase but also lipase has not been reported at all.

Therefore, an object of the present invention is to provide a digestive enzyme activity-inhibiting substance having an action of inhibiting not only the activity of α-amylase but also various digestive enzymes such as lipase easily and inexpensively. Another object of the present invention is to provide a diet food useful for suppressing hyperglycemia, hyperlipidemia and the like at low cost.

[0005]

According to the present invention, in order to achieve the above object, a digestive enzyme activity inhibitory substance obtained by filtering and purifying an extract of seaweed is provided. Furthermore, according to the present invention, there is provided a diet food containing a digestive enzyme activity inhibitor obtained by filtering and purifying the extract of seaweed.

[0006]

The present inventor has found that the extract from seaweed has not only a high α-amylase activity inhibitory action but also a lipase activity inhibitory action and can be easily obtained by an extraction procedure. They have found the present invention and completed the present invention. That is, the digestive enzyme (α-amylase, lipase) activity-inhibiting substance of the present invention comprises a seaweed extract. The seaweeds used in the present invention include brown algae (seaweed, kelp, squid, sardine, psyllium,
Mozuku, Honda straw, etc.), green algae (Anaosa, Aonori, mill, etc.), red algae (Ogonori, Hirakusa, Fukuronori, etc.) and the like. Above all, the activity of brown algae is extremely high.

The digestive enzyme (α-amylase, lipase) activity-inhibiting substance of the present invention can be obtained by extracting with a suitable solvent as it is untreated, except for appropriately cutting algae as described above. The extraction method, temperature, etc. are not particularly limited, but from the viewpoint of extraction efficiency, it is preferable to carry out the extraction by hot extraction of the solvent with seaweed. The reflux temperature varies depending on the solvent used, but the use of a temperature of 100 ° C. or higher does not impair the activity of the α-amylase inhibitor of the present invention.

As the solvent, in addition to water-soluble organic solvents such as water, alcohols, and acetone, aromatic hydrocarbons such as benzene and toluene, and halogens of aliphatic hydrocarbons such as chloroform and carbon tetrachloride. Compound and the like. Further, ethers, esters, ketones and the like can be used, but those having a low boiling point are preferable. In addition, water and alcohols are preferable from the viewpoint of safety and convenience in use after extraction. Examples of ether solvents include ethyl ether, isopropyl ether, dimethyldioxane, carbitol, cellosolves and the like. Examples of the ester solvent include ethyl acetate, propyl acetate, butyl acetate, methyl propionate, amyl formate, butyl formate, methyl lactate, methyl acrylate, ethyl acrylate and the like. Examples of the ketone solvent include acetone, diethyl ketone, methyl ethyl ketone, methyl butyl ketone, methyl propyl ketone and the like. Examples of the alcohol solvent include methyl alcohol, ethyl alcohol, propyl alcohol, butyl alcohol, amyl alcohol, acetonylmethanol and the like.

The digestive enzyme (α-
An aqueous solution containing an amylase, lipase) activity-inhibiting substance or an extract, if desired, is dried by a known method such as spray drying, freeze drying, or vacuum drying to obtain a powdery solid substance, which is directly used as a dieting agent. Alternatively, it may be added to a suitable food carrier, or may be formulated with a known auxiliary agent for liquid formulation, tableting or granulation. Known additives such as excipients, fillers, lubricants, binders, fragrances, and colorants can be used as auxiliaries for formulation of these. The solid substance containing the above-mentioned digestive enzyme (α-amylase, lipase) activity inhibitor is made into tablets or granules by the known tableting means or granulating means together with the above-mentioned auxiliaries.

The diet food containing the digestive enzyme (α-amylase, lipase) activity inhibitor thus obtained (in the present specification means a broad concept including the above-mentioned tablets, granules, etc.) is a food. It prevents the breakdown of starch and fat ingested as, and prevents it from being digested and absorbed as an energy source. Therefore, obesity, hyperglycemia, hyperlipidemia and the like can be effectively suppressed.

[0011]

EXAMPLES The present invention will be specifically described below with reference to examples. Example 1 From the edible seaweed, several representative seaweeds of each of the green algae, brown algae, and red algae listed in Table 1 were collected. Then, 100 g of untreated raw seaweed was crushed, and each was crushed together with 100 mL of the solvent shown in Table 1 to 30
After heating under reflux for a minute, the extract was filtered, the residue obtained by concentrating and drying the filtrate under reduced pressure was dissolved in 50 mL of ethanol, and the ethanol-soluble portion was filtered off. The extract was made into a 1% aqueous solution.

Test Example 1 (α-of extracts from various seaweeds)
Amylase activity inhibitory action) The degree to which the extract material obtained in Example 1 above inhibits α-amylase from hydrolyzing starch was measured. The test was carried out by comparing the presence or absence of α-amylase activity inhibition of the extract with water (blank test) as follows. First, 1 mL of water in 9 test tubes
Add 1 mL of 1% α-amylase aqueous solution to the first one and mix well, take 1 mL of it and mix in the next test tube, add 1 mL to the next test tube And mix. Repeat this operation in sequence,
Discard the last 1 mL. Then α-amylase 1
% Aqueous solution 0.5, 0.25, 0.12, 0.06, →
0.0019mL was taken and the total volume was 1mL with water.
The book is obtained. Attach one of the 1% aqueous solution of α-amylase without water. As described above, a set of 10 test tubes is prepared for the number of samples and a blank test and cooled in ice water. Then, 0.2 mL of each 0.1% aqueous solution of the seaweed extract obtained in Example 1 was placed in each test tube, while 0.2 mL of water was placed in each test tube as a blank test. Furthermore, add 5 mL of soluble starch to each test tube, and after adding all, add 40 ° C at a time.
Put it in the constant temperature bath and leave it for 30 minutes.

After a certain period of time, it is again put in ice water and cooled. Then, water is added from the upper end of each test tube up to about the width of the finger, and one drop of 0.1N iodine solution is added. Then, each test tube exhibits a color such as blue-violet, purple, red, and yellow, but among these, blue color is no longer observed, and a test tube exhibiting a purple color is taken, and 1 mL of the amount of α-amylase present in the test tube Calculate the number of mL of 1% starch solution decomposed by the α-amylase solution. For example, the one containing 0.02 mL of α-amylase solution turns purple in 30 minutes,
If the one with a smaller amount of α-amylase was blue, it means that 0.02 mL of the α-amylase solution was decomposed until the 1% starch solution 5 mL of the blue iodine reaction disappeared. Therefore, the amount of 1% starch liquid that 1 mL of α-amylase solution should decompose is 250 mL. Therefore, the starch-refining power of α-amylase is represented by the following formula 1.

[Equation 1] Therefore, if you examine the changes in the starch paste refining ability,
The presence or absence of inhibition of α-amylase activity is known.

The following Table 1 shows the test results of the degree of inhibition of the starch paste refining ability of the seaweed extract obtained as described above.

[Table 1] As is clear from Table 1, the extract from seaweed of the present invention has a high starch paste refining activity inhibition rate and an excellent α-amylase activity inhibitory action.

Test Example 2 (Lipase activity inhibitory action of extracts from various seaweeds) It was investigated whether the extract substance obtained in the above Example 1 inhibits the activity of lipase which is an enzyme that hydrolyzes fat. It was The test was carried out using a commercially available reagent for measuring lipase “BMY” (manufactured by Boehringer Mannheim). This is a method in which triolein is used as a substrate and the rate of decrease in the turbidity of this suspension is measured in the ultraviolet to determine the lipase activity. The composition of the kit is shown in Table 2 below.

[Table 2]

The test was carried out by using the 0.1% aqueous solution of the seaweed extract obtained in Example 1 and comparing the presence or absence of inhibition of lipase activity with water (blank test). First, the total amount of purified water in 1 bottle of buffer / substrate agent is 2
Add to 7.5 mL, dissolve the contents to make a test solution, and dissolve lipase in 1 mL of purified water to make a lipase aqueous solution. After preliminarily heating the test solution to 37 ° C., 2.5 mL was placed in a cuvette, and a 0.1% aqueous solution of seaweed extract (blank test was water) was added to the sample in a volume of 0.1 m.
L, 0.1 mL of an aqueous lipase solution was added, and the mixture was mixed without foaming, and the mixture was incubated in a constant temperature bath at 37 ° C. for 4 minutes, and then the absorbance was read.
After incubating in a constant temperature bath at 7 ° C (9 after mixing the sample
After a minute), read the absorbance. The presence or absence of inhibition of lipase activity can be determined from the difference in the amount of change in the absorbance. That is, if the amount of change in the absorbance is smaller than that in the blank test, it means that the activity of lipase is inhibited.

The test results of inhibition of lipase activity by the extract from seaweed in Test Example 2 are shown in Table 3 below.

[Table 3] As is clear from Table 3, the extract from seaweed of the present invention also has a lipase activity inhibitory action.

[0018]

INDUSTRIAL APPLICABILITY As described above, the extract of seaweed of the present invention has not only a high α-amylase activity inhibitory action but also a lipase activity inhibitory action, and can be easily obtained by an extracting operation. Therefore, this digestive enzyme (α
-Amylase, lipase) It is possible to provide a diet food containing an activity inhibitor at a low cost, and the obtained diet food prevents the decomposition of starch and fat ingested as food and digests and absorbs it as an energy source. Can be effectively prevented, and thereby obesity, hyperglycemia, hyperlipidemia and the like can be effectively suppressed.

Claims (3)

[Claims] 1. A digestive enzyme activity inhibitor obtained by filtering and purifying an extract of seaweed. 2. The digestive enzyme activity inhibitor according to claim 1, wherein the seaweed is brown algae, green algae or red algae. 3. A diet food containing a digestive enzyme activity inhibitor obtained by filtering and purifying an extract of seaweed.