WO2011033717A1

WO2011033717A1 - Goya koji composition

Info

Publication number: WO2011033717A1
Application number: PCT/JP2010/005064
Authority: WO
Inventors: 盛雄稲福; 恵与那覇; 早紀徳田
Original assignee: 株式会社琉球バイオリソース開発; 有限会社水耕八重岳
Priority date: 2009-09-15
Filing date: 2010-08-13
Publication date: 2011-03-24
Also published as: TW201114374A; JP2012236771A

Abstract

Disclosed are a goya (Momordica charantia L. var. pavel Crantz) koji composition, which is obtained by fermenting goya, that is an inexpensive and easily available material, with a koji mold, has elevated antioxidative and ACE-inhibitory activities compared with the activities before the fermentation, exerts improved functional effects compared with a product obtained by fermenting goya with a lactic acid bacteria, has a relieved bitterness and shows no side effect; and a method for producing the same. The goya koji composition having elevated antioxidative and ACE-inhibitory activities is obtained by adding a carbon source to dried and ground goya, adding water thereto, fermenting the mixture with a koji mold, i.e., Aspergillus oryzae (IFO4079) or Aspergillus awamori (IFO6082), and then drying the fermented matter. This koji composition shows significantly elevated antioxidative and ACE-inhibitory activities compared with a product obtained by fermenting goya with a lactic acid bacteria.

Description

Bitter gourd composition

The present invention relates to a bitter gourd composition, and more particularly, a bitter gourd or gourd pre-treated product fermented using koji molds to increase the antioxidant activity and ACE inhibitory activity and total polyphenol, The present invention relates to an antioxidant and an ACE inhibitor containing the goya composition as an active ingredient.

The action of antioxidants is to remove active oxygen generated in the body. This active oxygen is generated by the reaction of oxygen and various substances in the body, and causes a series of chain reactions and damages tissues in the body. It is said that. When exercising, it absorbs more oxygen than usual, so more active oxygen is produced, and it causes great damage to internal tissues. However, the living body has the ability to adapt to various environments and repairs it. However, if it cannot be repaired, it is said that cancer (cancer), aging, arteriosclerosis, myocardial infarction, and stroke are caused. It is said that prevention or treatment can be carried out to some extent by supplementing ingredients containing antioxidant activity with food or medicine.

Also, as one cause of hypertension, it is known that angiotensin I converting enzyme (hereinafter referred to as ACE), which controls the renin-angiotensin system, which is a pressor system, is deeply involved. By the action of ACE, the angiotensin I C-terminal dipeptide His-Leu is released to produce active angiotensin II. Whereas angiotensin I has no physiological activity, angiotensin II has a strong vasoconstrictive action, that is, a pressor action. ACE is also called kinase II, and acts on pressurization by degrading bradykinin, which has a strong vasodilatory action. Therefore, hypertension can be prevented or treated by inhibiting ACE having these actions.

Polyphenol is a general term for plant components having a plurality of phenolic hydroxyl groups (hydroxy groups) in the same molecule, and is contained in almost all plants, and the number is said to be over 5000 species. It is a water-soluble (partially fat-soluble) substance that has excellent antioxidant ability and is a plant pigment and bitter component produced by photosynthesis.

Polyphenols are the second largest component of dietary fiber in plant components and are important substances that contribute to the generation, activation, and division of plant cells related to biological defense and species preservation. It is said to have antimutagenicity, endocrine (hormone) promoting action and the like. Typical examples include anthocyanin, catechin, flavone, isoflavone, flavan, flavanone, chlorogenic acid, ellagic acid, lignan, curcumin, coumarin and the like.

In recent years, the number of patients with so-called lifestyle-related diseases such as diabetes, hyperlipidemia, and hypertension has increased due to the westernization of eating habits and lack of exercise accompanying lifestyle changes, which has become a major social problem. Under such circumstances, prevention and treatment of lifestyle-related diseases are urgently needed, and effective foods to be eaten on a daily basis are required to improve these diseases.

Goya is called the Japanese name “Tsurureishi” and is generally called “Nigauri”. Scientific name Momordica charantia L.var.pavel Crantz, which is rich in vitamin C and contains carotene, vitamin B, vitamin K, potassium, magnesium, iron, phosphorus, dietary fiber, etc. Although it is a plant that has been eaten, in recent years it has a health-oriented boom and plants that are easy to grow, and it has been widely cultivated in places other than Okinawa and has become a daily ingredient during the season.

Traditionally, many plants that produce natural effects when eaten as they are are eaten as they are, and those that dry the plants and drink like tea and ingest the contained active ingredients are known. Recently, by making microorganisms act on plant raw materials, it has become easier to drink. For example, with regard to bitter gourd, a method for producing bitter tea that cuts bitter melon, fermented by adding lactic acid bacteria, and then dried (see, for example, Patent Document 1), lactic acid fermentation, citric acid fermentation, alcohol fermentation to bitter gourd A fermented product obtained by performing microbial fermentation such as acetic acid fermentation (for example, see Patent Document 2) is known.

On the other hand, a goya koji composition that is fermented by using koji mold as a goya or its pre-treated product and has enhanced antioxidant activity and ACE inhibitory activity and a method for producing the same are not known.

JP 2003-169621 A JP 2006-75083 A

The present invention is an economically inexpensive and easily obtainable goya that has an increased antioxidant activity and ACE inhibitory activity as compared with that before fermentation by koji mold, and an increase in total polyphenols, and also compared with fermentation by lactic acid bacteria. However, the present invention is to provide a bitter gourd composition that has high side effects and has reduced side effects and has no side effects, and a method for producing the same.

Many of the plants that grow in Okinawa are known to be effective in preventing or treating diseases, and the plants have been dried, decocted, or used daily for cooking, etc. The present inventors have further extended the study to other plants, tried various processing treatments on plants known to contain physiologically useful ingredients, and maximized the effective ingredients contained therein. We have made every effort to make it available to the limit. As for goya, in addition to fermentation by lactic acid bacteria, in addition to fermentation by lactic acid bacteria, it was found that as a result of the action of various other microorganisms, gonorrhea treatment increased antioxidant activity and ACE inhibitory activity. It came to be completed.

That is, the present invention is (1) the following (a) hot water extract at 80 ° C. obtained by fermentation treatment with koji molds of goya or a pre-processed product of goya has an antioxidant activity enhanced by fermentation treatment with koji molds. And (b) a 80% ethanol extract has an ACE inhibitory activity enhanced by fermentation treatment with Aspergillus oryzae; (2) Aspergillus oryzae (Aspergillus oryzae) Or the bitter gourd composition as described in (1) above, which is Aspergillus awamori, or (3) Aspergillus oryzae (IFO4079) or Aspergillus awamori (IFO6082) (2) the bitter gourd composition described in (2) above, and (4) the total poly, measured by the foreign dennis method. The bitter gourd composition according to any one of (1) to (3) above, wherein (5) the above (1) to (4) A method of using the bitter gourd composition according to any one of the above for the preparation of an antioxidant or an ACE inhibitor, or (6) an effective bitter gourd composition according to any one of (1) to (4) above An antioxidant as an ingredient, (7) an ACE inhibitor containing the bitter gourd composition according to any one of (1) to (4) as an active ingredient, and (8) a carbon source for dried and ground bitter gourd A gourd characterized in that it includes a fermentation step for fermenting using gonococcus to increase the antioxidant activity and ACE inhibitory activity, and a drying step for drying the treated product after fermentation. (9) The carbon source is molasses (8) The method for producing a bitter gourd composition according to (8) above, or (10) the total amount of polyphenols in the bitter gourd composition as compared with that before fermentation (8) ) Or (9), or (11) the total polyphenol amount measured by the foreign dennis method is increased by 30% or more compared to before fermentation (10) This relates to a method for producing the bitter gourd composition.

INDUSTRIAL APPLICABILITY According to the present invention, fermented processing of gourd gourd of goya, which has higher antioxidant activity and ACE inhibitory activity, mitigates bitterness, is safer and can be obtained economically and economically than fermented processed products of goya lactic acid bacteria As a result, a supplement, food, or food material containing the goya koji composition is useful as an antioxidant composition or an ACE inhibitor composition.

The radical residual rate of the hot water extract of the bitter gourd fermented product by Aspergillus oryzae 4079 and 6082 of the present invention, calculated using the DPPH radical scavenging activity method, is shown together with before fermentation and control. The ACE inhibitory activity is shown for the post-fermentation increase in the ethanol extract of the bitter gourd processed product by Aspergillus oryzae 4079, 6082 of the present invention, together with that before fermentation. The radical residual rate of the hot water extract of the bitter gourd fermented product by Aspergillus oryzae 4079 and 6082 of the present invention calculated using the DPPH radical scavenging activity method is shown together with the lactic acid fermented product, before the aspergillus fermentation, and the control. The increase in ACE inhibitory activity after fermentation in the ethanol extract of bitter gourd fermented product by Aspergillus oryzae 4079, 6082 of the present invention is shown together with the pre-fermented and lactic acid fermented product. The chromatogram which shows the result by the HPLC analysis of the total polyphenol content in the hot-water extract of the processed goya before fermentation. The chromatogram which shows the result by HPLC analysis of the total polyphenol content in the hot-water extract of the bitter gourd processed material by the koji mold 4079 of this invention. The chromatogram which shows the result by HPLC analysis of the total polyphenol content in the hot-water extract of the bitter gourd processed material by the koji mold 6082 of this invention. The chromatogram which shows the result by HPLC analysis of the total polyphenol content in the hot-water extract of the bitter fermented product by lactic acid bacteria. The total polyphenol content in the methanol extract of the pre-fermentation product calculated using the foreign dennis method, the koji mold 4079 fermentation process product of the present invention, the koji mold 6082 fermentation process product of the present invention, and the lactic acid bacteria fermentation process product is shown.

As the bitter gourd composition of the present invention, (a) a hot water extract at 80 ° C. obtained by fermenting gourd or goya pre-treated product with gonorrhoeae has an antioxidant activity enhanced by the fermenting process using gourd. Physical properties that the hot water extract at 80 ° C. has enhanced antioxidant activity compared to the unfermented processed product; and (b) the 80% ethanol extract is fermented by Aspergillus oryzae. Physical properties with enhanced ACE inhibitory activity upon treatment; ie, 80% ethanol extract derived from bitter gourd with physical properties having enhanced ACE inhibitory activity relative to the unfermented treated counterpart The composition is not particularly limited as long as it is a composition. Examples of the pretreatment of bitter gourd include bitter gourd treatment, shredding treatment, drying treatment, ultrasonic homogenization treatment, freezing treatment, and the like. , These may be combined process of two or more. Preferable examples include a method of chopping goya as it is and then drying, or a method of chopping or pulverizing a processed product that has been previously subjected to a drying treatment, a freezing treatment, or the like. In addition, as a method for producing the bitter gourd composition of the present invention, a carbon source is added to dried and ground bitter gourd, hydrolyzed, and fermented using koji molds to increase antioxidant activity and ACE inhibitory activity. There is no particular limitation as long as the method includes a fermentation step for drying and a drying step for drying the processed product after fermentation. Specifically, the dried and pulverized product is, specifically, dried with goya set at about 40 ° C. to 100 ° C. Examples of the processed products are those that are shredded or crushed to 0.05 to 1 mm after drying treatment such as heat drying treatment indoors or sunlight drying treatment outdoors.

The koji mold used in the present invention is not particularly limited, but yellows such as Aspergillus oryzae, Aspergillus flavos, Aspergillus polyoxogenes, Aspergillus sojae, etc. Aspergillus awamori, Aspergillus kawauchii, Aspergillus usami, Aspergillus niger, Monascusanka, Monascusanka Examples include Monascus purpureus. In particular, in the present invention, Aspergillus oryzae and Aspergillus awamori are preferable, and Aspergillus oryzae IFO 4079 or Aspergillus awamori IFO6082 is more preferable.

As fermentation conditions of the present invention, it is possible to efficiently ferment by treating raw materials, adjusting fermentation temperature, fermentation time, pH, water content suitable for growth of Aspergillus and selecting the type of fermentation substrate. Depending on the progress of fermentation and preferences, it can be changed and selected as appropriate. For example, for the treatment of bitter gourd, the dry powder of bitter gourd described above, for example, can be used. The fermentation temperature is 28 to 32 ° C., particularly 30 ° C., and 72 to 168 hours is appropriate, pH 4 to 7, The raw material powder is preferably 2.5 to 5 times (weight), particularly about 3 times. It is preferable to add a carbon source or a nitrogen source to a fermentation substrate such as dried or ground bitter gourd. As the carbon source, sugars such as molasses, glucose and sucrose are preferable, and these carbon source and nitrogen source are 1 You may use a seed | species individually or in mixture of 2 or more types. The amount of saccharide added is preferably 1 to 5% by weight per medium (excluding water). The nitrogen source is preferably rice bran, bran or the like, and the addition amount is preferably 1 to 10% by weight per medium (excluding water), and particularly about 3% by weight is appropriate. As for the koji mold, it is preferable to inoculate 5 to 50 ml / 100 g of fermentation substrate as a spore suspension (10 ⁵ to 10 ⁸ / ml) in fermentation. After fermentation, it is preferable to perform a fermentation stop treatment from the viewpoint of preventing overfermentation. As such a fermentation stop treatment, a method of exposing to steam at 80 to 100 ° C. for 30 minutes can be specifically exemplified.

After the fermentation stop treatment, drying is preferably performed by a dryer so that the moisture value is 10% by weight or less. As a drying method, it can be performed by heat drying or freeze drying. Is preferably carried out at 100 ° C. or lower because the deactivation of the physiologically active ingredient can be prevented.

After drying, if necessary, sterilization is performed by a known method such as heating to obtain the bitter gourd composition of the present invention. The bitter gourd composition is used as a food material as it is or as a compounding agent that can be used by adding to food. Moreover, you may extract an active ingredient from a fermentation processed material with a solvent.

As the solvent used for the solvent extraction treatment, for example, any of an aqueous medium and an organic solvent may be used, and the solvent may be used alone or as a mixed solvent in which a plurality of solvents are combined. Examples of the aqueous medium include water, purified water, deionized water, distilled water, and warm water and hot water thereof. Examples of the organic solvent include lower alcohols such as methanol, ethanol, propanol, isopropanol, isobutyl alcohol, and t-butyl alcohol and their hydrates, liquid polyhydric alcohols such as glycerin, propylene glycol, and 1,3-butylene glycol, and their hydrates. Products, alkyl acetates such as methyl acetate and ethyl acetate, aliphatic ketones such as acetone and ethyl methyl ketone, aliphatic ethers such as dimethyl ether and diethyl ether, aliphatic hydrocarbons such as hexane, heptane and petroleum ether, and fats such as cyclohexane Cyclic hydrocarbons, aromatic hydrocarbons such as toluene and benzene, halogenated aliphatic hydrocarbons such as chloroform, dichloromethane, 1,1,1,2-tetrafluoroethane, 1,1,2-trichloroethene, sesame oil, Corn oil, Leave oil, it may be mentioned edible fat and oil of cotton seed oil, methane, ethane, propane, propylene, butane, liquefied aliphatic hydrocarbons butylene, dimethyl sulfoxide and the like. Preferable extraction methods include, for example, a method in which hot water at 80 to 100 ° C. is used for 30 minutes to 24 hours, and an alcohol extraction method. In the case of the alcohol extraction method, it is preferable to use ethanol having an ethanol concentration of 60 to 80% by volume. In this case, a method of extracting at 0 to 80 ° C. for 30 minutes to 5 days can be exemplified. The method of filtering etc. can be mentioned.

The bitter gourd composition which is a fermented processed product of gourd or the processed product of goya of the present invention by Aspergillus oryzae has an action of improving antioxidant activity and ACE inhibitory activity. Therefore, the present invention relates to a method of using the bitter gourd composition of the present invention for the preparation of an antioxidant or an ACE inhibitor, an antioxidant containing the bitter gourd composition of the present invention as an active ingredient, The present invention also relates to an ACE inhibitor containing a goya koji composition as an active ingredient. Moreover, the food containing the bitter gourd composition of the present invention can be distributed and sold as a functional food or food material with a label indicating that it is used for improving antioxidant activity and ACE inhibitory activity.

The bitter gourd composition of the present invention has higher antioxidant activity and ACE inhibitory activity effect than those before and after lactic acid fermentation, and the total polyphenols when measured by the foreign dennis method A gallic acid equivalent of 350 mg% or more, preferably 400 mg% or more, more preferably 450 mg% or more, and is a koji composition that alleviates bitterness unique to bitter gourd and feels delicious. The composition is extracted with hot water to make a drink, or added to various foods and drinks so as not to impair the taste of the added foods and drinks, for improving antioxidant activity and for improving ACE inhibitory activity Food or food material can be produced. Examples of foods and beverages to be added include noodles, rice cakes, cookies, cereals, confectionery (snacks), sprinkles, processed livestock products, processed fish products, canned foods, juices, soft drinks, nutritional drinks, etc. Can be mentioned.

Further, the antioxidant and ACE inhibitor containing the bitter gourd composition of the present invention as an active ingredient can be used as a medicine in addition to supplements and foods, in which case, a pharmaceutically acceptable normal carrier, Various preparation compounding components such as a binder, a stabilizer, an excipient, a diluent, a pH buffer, a disintegrant, a solubilizer, a solubilizer, and an isotonic agent can be added. Such pharmaceutical dosage forms are generally used dosage forms such as powders, granules, capsules, syrups and suspensions, orally, or for example solutions, emulsions, suspensions, etc. It can also be made into the dosage form. The amount of intake varies depending on the human body weight, age, sex, symptoms, etc., and the dose per day is determined according to the symptoms, but can be taken for a long time.

In the present invention, when the bitter gourd composition is used daily as an antioxidant or as an ACE inhibitor, the extract or powder extracted therefrom can be used for tea bags, plastic bottles, cans, and drinks. . Moreover, it can also be set as a granular sprinkling. Furthermore, beverages in which extracts and granules extracted from the bitter gourd composition of the present invention are dissolved in drinking water, juice, etc., baked confectionery such as bread, cake, rice cracker, Japanese confectionery such as mutton, frozen confectionery, chewing gum, jelly, etc. Bread, confectionery, noodles such as udon, soba, fish paste products such as kamaboko, ham, fish sausage, seasonings such as miso, soy sauce, dressing, mayonnaise, sweetener, cheese, butter, yogurt, ice cream It can be added to dairy products such as puddings, and various side dishes such as tofu, konjac, and other boiled sardines.

In the bitter gourd composition of the present invention, it is important that the antioxidant activity is enhanced before fermentation. The strength of the antioxidant activity can be measured by β-carotene method, DPPH method, rhodan iron method and the like. Here, the β-carotene method is a measurement method using β-carotene that fades and becomes transparent by being easily oxidized, and a mixture of β-carotene, linoleic acid, and a sample is auto-oxidized, This is a method for measuring the strength of antioxidant activity by measuring the degree of fading. If the yellow color of β-carotene does not fade, it can be determined that the sample has antioxidant activity. The DPPH method is a method based on the measurement of radical scavenging ability using 2,2-diphenyl-1-picrylhydrazyl. The rhodan iron method is a spectrophotometer for measuring the degree of auto-oxidation of linoleic acid ( 500 nm).
About the bitter gourd composition of this invention, it is preferable that the antioxidant activity measured by the hot water extraction method is strengthened 30% or more, especially 40% or more compared with before fermentation processing.

Moreover, the bitter gourd composition of the present invention has improved ACE inhibitory activity as compared with the unfermented processed product. This is due to the fact that the ACE (Angiotensin I Converting enzyme) inhibitory activity of the fermented processed product obtained by the present invention is higher than that of the unfermented processed product. ACE is an angiotensin I converting enzyme related to the conversion of angiotensin I to angiotensin II. Angiotensin II is a hormone that increases blood pressure, and thus it can prevent an increase in blood pressure by inhibiting the activity of ACE. It becomes possible. The bitter gourd processed product is preferably increased by 20% or more, particularly 30% or more from that before fermentation. The ACE inhibitory activity was measured in 80% ethanol extract. Hippuryl L-, which is a hippuric acid derivative dissolved in borate buffer containing sodium chloride, was added to the ACE. This is a method in which the amount of hippuric acid liberated by adding histidylcineL-leucine is measured and determined as the production ratio when water is used as a control.

Furthermore, the method for producing the bitter gourd composition of the present invention is characterized in that the total amount of polyphenols is increased compared with that before the fermentation, and the total amount of polyphenols measured by the foreign dennis method is 30% compared with that before the fermentation. As mentioned above, a method of increasing preferably 40% or more, more preferably 50% or more, and especially 60% or more can be suitably exemplified. The enhancement of the antioxidant activity and the ACE inhibitory activity in the bitter gourd composition of the present invention is considered to be due to the production of polyphenol compounds in the koji fermentation (koji making) process.

The total polyphenol in the fermented product of the present invention can be measured by HPLC analysis using predetermined analysis conditions and by the foreign dennis method using a foreign dennis reagent.

More importantly, the bitter gourd composition of the present invention has higher antioxidant action and ACE inhibitory activity than the conventional processed bitter gourd fermented with lactic acid bacteria.

Hereinafter, the present invention will be described with reference to examples, but the present invention is not limited to the following examples.

Raw goya is sliced to a thickness of 1 to 5 mm, the sliced goya is dried in a drying chamber at 40 to 60 ° C. to a water content of 10% or less, and the diameter is reduced to 0. 0 mm by a pulverizer (Oster Blender, Oster). A pulverized product of 5 mm or less was obtained. A suspension prepared by mixing 30 g of dried bitter gourd powder, 0.9 g of molasses and 90 ml of water, adjusted to pH 5 and added aspergillus oryzae (IFO4079) spores to 10 ⁶ per ml of the suspension, Fermented at 30 ° C. for 168 hours. Thereafter, the fermentation was stopped by exposure to steam at 90 ° C., followed by drying until the water content became 10% by weight or less, and sterilization (130 ° C. steam, 5 to 15 seconds) was performed to obtain 31 g of bitter gourd powder.

Example 1 Except for using Aspergillus oryzae (IFO6082) in place of Aspergillus oryzae (IFO4079), Example 1 was carried out to obtain 31 g of bitter gourd powder.

In order to observe the growth state of koji in the fermentation processes of Examples 1 and 2 and to examine the number of viable bacteria, 1 ml of fermentation substrate was added to a standard plate agar medium (“Standard Agar Medium” manufactured by Eiken Chemical Co., Ltd., pH 7 to I went to 7.2). The appearance of the cocoon was confirmed by its appearance, etc., and the number of bacteria was examined for general viable bacteria. Table 1 shows the results of the number of general viable bacteria before fermentation, on the third day of fermentation, on the seventh day of fermentation, and on the ninth day of fermentation.

As for the growth of Aspergillus oryzae (IFO4079), the formation of cocoons can be confirmed as a whole from the second day, and aspergillus awamori (IFO6082) can be confirmed from the first day. Then, it was able to confirm the formation of wrinkles overall. As shown in Table 1, it was found that the number of viable bacteria increased with the passage of fermentation time when Aspergillus oryzae (IFO4079) and Aspergillus awamori (IFO6082) were used.

[Measurement of antioxidant activity by DPPH method]
1 g each of the dried powder of bitter gourd fermented product obtained in Examples 1 and 2 and the dried bitter gourd powder before fermentation was taken up to 50 ml with hot water at 80 ° C., and the liquid was used as a stock solution. Further, ascorbic acid as a positive control, a solution whose concentration was adjusted to 1 mg / ml was used as a stock solution to be measured. These stock solutions were measured according to the following DPPH method. 0.95 mL of 0.05 M Tris-HCl buffer (pH 7.4) (manufactured by WAKO), 1.0 mL of 0.1 mM DPPH-ethanol solution (manufactured by WAKO), and 100% ethanol. 0 mL was mixed and stirred. 0.05 ml of the previous stock solution was added to the reagent thus obtained. Absorbance (517 nm) was measured before the addition of the reagent and 30 seconds after the addition. Distilled water was used as a control, and the difference in absorbance before and after the addition of the reagent to the sample was calculated as DPPH radical consumption rate (%), where the difference in absorbance before and after the addition of the control reagent was 100. %) Subtracted from 100 was defined as DPPH radical residual rate (%). The results are shown in FIG.

As apparent from FIG. 1, the fermented bitter gourd obtained in Example 1 (sometimes abbreviated as “4079”) and Example 2 (sometimes abbreviated as “6082”) is a DPPH method. The antioxidant activity (DPPH radical residual rate) measured by the above was enhanced by 15% or more compared to the dried goya powder before fermentation.

[Measurement of ACE inhibitory activity]
The 80% ethanol extract of bitter gourd powder obtained in Examples 1 and 2 was dissolved in 50% dimethyl sulfoxide (DMSO) to a concentration of 20 mg / mL, and used for the test. After 15 μL of each sample and 50 μL of ACE were mixed, preincubated at 37 ° C. for 5 minutes, and then 125 μL of a substrate (Hippuryl-L-histidyl-L-leucine) dissolved in borate buffer containing sodium chloride was added at 37 ° C. The reaction was allowed for 30 minutes. After the reaction, 20 μL of 10% TFA was added to stop the reaction. After stopping the reaction, the amount of hippuric-acid generated by HPLC was measured, and the ACE activity was 100% using water as a control. The ratio is calculated, and the values subtracted from 100 are shown in FIG. 2 as ACE inhibitory activity (%).

As is clear from FIG. 2, it is found that the fermented bitter gourd has significantly higher ACE inhibitory activity (%) than before fermentation, indicating that it is about 40% higher than that before fermentation. ing.

[Comparative Example 1]
In Example 1, instead of Aspergillus oryzae (IFO4079), Streptococcus faecalis (IFO3128), Lactobacillus plantarum (IFO14713) and Lactobacillus casei3 (IFO1515) (IFO1515) The same procedure as in Example 1 was carried out except that the total number of mixed bacteria was 10 ⁶ / ml, and 31 g of fermented powder was obtained.

[Comparative test on antioxidant activity]
The bitter gourd lactic acid fermentation powder obtained in Comparative Example 1 was measured in the same manner as the antioxidant activity measurement test by the DPPH method. The results are shown in FIG. 3 together with the fermented koji powder obtained in Example 1 (6082) and Example 2 (4079) of the control and the present invention.

As is clear from FIG. 3, the activity obtained in the hot water extract was confirmed to be higher in the one obtained by the koji mold fermentation of the present invention than the goya lactic acid bacteria fermentation.

[Comparative test on ACE inhibitory activity]
The bitter gourd lactic acid fermentation-treated powder obtained in Comparative Example 1 was measured in the same manner as in the ACE inhibitory activity measurement test. The results are shown in FIG. 4 together with the fermented fermented powder obtained in the control, Example 1 (6082) and Example 2 (4079) of the present invention.

As is clear from FIG. 4, it was confirmed that the ACE inhibitory activity was increased in the ethanol extract obtained from the gourd lactic acid bacteria fermentation by the koji mold fermentation.

[Sensory test]
A sensory test was performed on the processed fermented bitter gourd by Aspergillus. Take 5 g of each of the fermented powder and fermented bitter gourd obtained in Example 1 (4079), the fermented powder obtained in Example 2 (6082) and the pre-fermented bitter gourd powder and take 500 ml of hot water for 5 minutes. Boiled and made tea. Ten panelists sampled these teas. Table 2 shows the number of people who answered that they were delicious.

As shown in Table 2, an improvement in palatability was confirmed after the fermentation of bitter gourd by Aspergillus. In particular, all 10 of the panelists who drank tea containing the fermented bitter gourd processed with Aspergillus oryzae (IFO 4079) of Example 1 answered that it was delicious, the one before fermentation was zero, and goya It has been found that the fermentation by Aspergillus oryzae reduces the bitterness and rather is effectively related to the deliciousness. In addition, even those using Aspergillus awamori (IFO6082) of Example 2 answered that 8 people were delicious in the fermented goya tea, indicating that it can be used as a beverage in substantially the same manner as in Example 1. Yes.

2 g of fermented bitter gourd fermented with koji mold obtained in Example 1 was stored in a tea bag to obtain an instant tea bag. The tea bag is put in 200 cc of hot water, extracted for 3 to 5 minutes, and then used for drinking.

10 g of gourd fermented powder of koji mold obtained in Example 2 is mixed with 100 g of sprinkled raw material consisting of bonito, white sesame and chopped seaweed in an appropriate mixing ratio, seasoned with sugar, soy sauce, salt and bonito extract, and sprinkled powder Got. Can be sprinkled on rice or chopped with leek to make instant soup.

[Polyphenol content measurement test by HPLC]
1 g each of the dried powder (4079 and 6082) of the bitter gourd processed with gonorrhea obtained in Examples 1 and 2, the dried powder of the processed bitter gourd before fermentation, and the powder processed with lactic acid fermentation obtained in Comparative Example 1 were measured. 25 ml of 80% methanol was added and homogenized for 1 minute. Thereafter, the mixture was centrifuged at 8000 rpm for 15 minutes, and the supernatant was filtered. To the remaining residue, 25 ml of 80% methanol was added again, homogenized for 1 minute, and filtered. The obtained filtrate was quantified to 50 ml and used as a measurement sample. HPLC analysis was performed under the following conditions.
Column: Develosil ODS-HG-5 (250 mm x inner diameter 4.6 mm Nomura Chemical)
Mobile phase: A solution; 10 mM phosphoric acid aqueous solution B solution; methanol / acetonitrile / water = 9/9/2
Gradient analysis flow rate: 1 mL / min so that liquid A becomes 55% after 40 minutes from 95% liquid A
Wavelength: UV (280nm)
Injection volume: 10 μL
In the hot water extract of each dry powder of the gourd fermentation processed product (Koji mold 4079 and 6082) by Aspergillus obtained in Examples 1 and 2 and the lactic acid fermented processed product obtained in Comparative Example 1 before fermentation The results of HPLC analysis are shown in FIGS.

As shown in FIG. 5, the peak area in the 10.7 minute range before the fermentation was 81843 mAU, whereas in FIG. 6, fermentation with Aspergillus (4079) showed 563039 mAU, and in FIG. 6082) showed 1909224 mAU, and an increase in the total phenol content could be confirmed. Moreover, as shown in FIG. 8, increase was not able to be confirmed by lactic acid fermentation.
In this way, the extract of bitter gourd fermented with koji has a higher total polyphenol content than the one before fermentation and the extract of fermented with lactic acid bacteria. It can be seen that there is a higher antioxidant effect.

[Total polyphenol content measurement test by the foreign dennis method]
1. Extraction Method 1 g of the sample obtained in Example 1 (Koji mold 4079), Example 2 (Koji mold 6082), and Comparative Example 1 was weighed, and 25 ml of 80% methanol was added and homogenized for 1 minute. Thereafter, centrifugation was performed at 8000 rpm for 15 minutes, and the supernatant was filtered. The remaining residue was again added with 25 ml of 80% methanol, homogenized for 1 minute, and filtered. The obtained filtrate was quantified to 50 ml and used as a measurement sample.
2. Foreign Dennis Method The total polyphenol content of the obtained extract was measured using the foreign dennis method. That is, 150 μL of the sample solution, 75 μL of foreign dennis reagent (FURUKA), and 75 μL of 2.5% sodium carbonate solution were respectively added to a 96-well microplate and stirred. The mixture was reacted at room temperature for 60 minutes, and then the absorbance at 750 nm was measured with a microplate reader. A calibration curve was prepared using gallic acid as a standard substance, and the total amount of polyphenols was calculated. The results are shown in Table 3 and FIG.

As shown in Table 3 and FIG. 9, as the gallic acid equivalent, an increase in the total polyphenol content was confirmed by gonococcal fermentation (6082 and 4079), which was 1.6 times and 1.3 times that before fermentation, respectively. . From these values, it can be seen that those obtained by Aspergillus oryzae fermentation increase the total polyphenol content by 30% or more and 60% or more, respectively, compared to before fermentation. Moreover, the same increase was not confirmed by lactic acid fermentation.

Claims

A bitter gourd composition having the following physical properties (a) and (b), which is obtained by fermenting bitter gourd or a bitter gourd pretreatment product with Aspergillus oryzae.
(A) a hot water extract at 80 ° C. has an antioxidant activity enhanced by fermentation treatment with Aspergillus oryzae;
(B) 80% ethanol extract has ACE inhibitory activity enhanced by fermentation treatment with Aspergillus oryzae;
2. The bitter gourd composition according to claim 1, wherein the Aspergillus is Aspergillus oryzae or Aspergillus awamori.
The goya koji composition according to claim 2, wherein the koji mold is Aspergillus oryzae (IFO4079) or Aspergillus awamori (IFO6082).
The bitter gourd composition according to any one of claims 1 to 3, wherein the total amount of polyphenols measured by the foreign dennis method is 350 mg% or more in terms of gallic acid equivalent.
A method of using the bitter gourd composition according to any of claims 1 to 4 for the preparation of an antioxidant or an ACE inhibitor.
An antioxidant comprising the bitter gourd composition according to any one of claims 1 to 4 as an active ingredient.
An ACE inhibitor comprising the bitter gourd composition according to any one of claims 1 to 4 as an active ingredient.
Adding a carbon source to dried and ground bitter gourd, hydrolyzing it, fermenting it using koji molds, increasing the antioxidant activity and ACE inhibitory activity, and drying the dried processed product And a method for producing a bitter gourd composition.
The method for producing a bitter gourd composition according to claim 8, wherein the carbon source is molasses.
The method for producing a bitter gourd composition according to claim 8 or 9, characterized in that the total amount of polyphenol in the bitter gourd composition is increased compared to that before fermentation.
The method for producing a bitter gourd composition according to claim 10, wherein the total amount of polyphenols measured by the foreign dennis method is increased by 30% or more compared with that before fermentation.