KR102110363B1 - A composition comprising maclurin as an active ingredient for inhibiting food browning - Google Patents

Abstract

The present invention relates to a composition for preventing food browning, which includes macrorin as an active ingredient.
The composition comprising the macrorin of the present invention as an active ingredient can suppress browning of fruits or vegetables that are easy to brown for a long time, and when added directly, has an antioxidant activity enhancing effect. In particular, since it was used as a control, vitamin C and arbutin had a superior anti-browning effect as well as excellent anti-oxidant effect, the composition containing macrorin as an active ingredient suppressed browning and increased the antioxidant capacity of foods, resulting in quality Productivity can be improved.

Description

A composition comprising maclurin as an active ingredient for inhibiting food browning}

The present invention relates to a composition for preventing food browning, which includes macrorin as an active ingredient.

Browning is a phenomenon in which food turns brown in the process of storage, processing, and cooking of food. There are cases where enzymes are involved in browning (enzymatic browning) and cases where enzymes are not involved (non-enzymatic browning). Browning not only impairs the appearance of food and deteriorates its aroma, but also results in increased damage to producers by deteriorating the product's commerciality, such as nutritionally losing or reducing vitamins and essential amino acids.

As part of efforts to prevent such browning, a method of using various additives to fruits or removing solids by ultrafiltration, electrodialysis, etc. is being sought (Monsalve GA, et al., 1995; Kang HA, et al. , 1998), in terms of business application, there are still many points that are still insufficient.

For example, the heat treatment, such as scalding or peeling, which is most effective in preventing browning, may adversely affect the texture, aroma, and taste of the fruit on the other hand, and the heating conditions should be sufficiently considered. In addition, ascorbic acid is widely used as an anti-browning agent and antioxidant, but ascorbic acid itself causes browning by an oxidase, an addition amount or a coexisting material must be considered.

Many of the “browning” inhibitors developed so far have been used as a preventer to prevent browning of flesh in the process of harvesting and processing fruits. Vitamin C, reducing agent, acidulant, chelating agent, etc. have been traditionally used, but safety There is a case where this effect is concerned or when it is directly added to food, the effect is insignificant, and thus a better anti-browning agent is needed.

Thus, the present inventors completed the present invention by confirming that as a result of earnest research to develop an anti-browning agent having an excellent effect, macurin has an excellent anti-browning effect and enhances the antioxidant activity of food.

One object of the present invention is to provide a composition for preventing food browning, which includes macrorin as an active ingredient.

Another object of the present invention is to provide a food composition comprising the composition for preventing food browning.

Another object of the present invention is to provide a method for preventing food browning, comprising the step of treating the composition with food.

One aspect of the present invention for achieving the above object, to provide a composition for preventing food browning, comprising as an active ingredient.

In the present invention, the term "maclurin (maclurin)" refers to a compound having the structure of Formula 1 below.

[Formula 1]

The physiological activity of macrorin is known to inhibit the metastasis or invasion of cancer cells (Ku, Min Jung, et al,) 2015). However, the effect of preventing food browning of macurine was first identified by the present inventors.

The macrorin may be purchased, directly extracted, or separated.

In the present invention, the term "food" may mean any ingestible food that is susceptible to browning. The food may be fruits, vegetables, meat, dairy products, bread, but is not limited thereto.

In the present invention, the term, "browning" is a phenomenon in which food turns brown in the process of storage, processing, and cooking of food. When browning involves an enzyme (enzymatic browning) and when no enzyme is involved (non-enzymatic) Browning).

The substrates for enzymatic browning are phenols and polyphenols in plants, and quinones with high reactivity are produced by the action of enzymes such as catechol oxidase and laccase. The resulting quinones are polymerized by themselves or an amino compound and a reactant are polymerized to form melanoidin, a colored polymerization material.

The non-enzymatic browning is called the Maillard reaction after the discoverer and the aminocarbonyl reaction after the atomic group involved in the reaction. In general, browning is unfavorable, such as browning of dairy products, browning of bread and biscuits, browning of fats and oils, browning of potatoes, browning of tea, browning of fruit and dried fruits, browning of meat, etc. As seen in miso, soy sauce, etc., there are cases in which desirable properties are given to foods, such as the generation of aroma components, coloring, and expression of antioxidant properties.

The composition may be to inhibit polyphenol oxidase.

In a specific embodiment of the present invention, the potato soup was treated with macurine and the absorbance according to browning was measured to confirm the inhibitory activity of the polyphenol oxidase, confirming that the inhibitory activity of the group treated with macrorin was excellent. (Fig. 3).

The composition according to the present invention may be added to food to enhance the antioxidant capacity of food.

The antioxidant refers to the action of inhibiting oxidation, and the human body has a balance of an oxidant and an antioxidant (antioxidant), but due to various factors, this balance is unbalanced and promotes oxidation. When tilted, oxidative stress is induced, causing potential cell damage and pathological diseases. Reactive oxygen species (ROS), which are the direct cause of these oxidative stresses, are unstable and highly reactive, so they can easily react with various biological materials, attack the polymers in the body, cause irreversible damage to cells and tissues, or mutate, It causes cytotoxicity and carcinogenesis. Reactive nitrogen species (RNS) such as NO, HNO ₂ and ONOO ^- are produced in large quantities due to the immune response of macrophage neutrophils and other immune cells during an inflammatory reaction, and ROS is also generated. The free radicals as described above oxidize and destroy cells in the body, and thus are exposed to various diseases. Therefore, adding the macrorin of the present invention to food can increase the antioxidant capacity of the food and contribute to health promotion.

In a specific embodiment of the present invention, as a result of measuring the ROS level according to the treatment of macrorin, ROS increased rapidly after treatment with SIN-1, a source of active oxygen production, but concentration in the group treated with SIN-1 and macrorin Independently, it was confirmed that the amount of free radicals decreased (FIG. 4A).

In another specific embodiment of the invention, the mark ONOO according to the process of the rurin ^- As a result of measuring a scavenging active oxygen generated cause SIN-1 after treatment ONOO ^- but production is increased, in the group treated with the mark rurin ONOO ^- It was confirmed that the production was significantly reduced (FIG. 4C).

There is no particular limitation on the type of food to which the composition containing the above-mentioned macrorin can be added as an active ingredient, for example, various drinks, gums, teas, vitamin complexes, health supplements, and the like. Other ingredients that do not interfere with the food browning prevention effect may be added to the food, and the type is not particularly limited. For example, as a conventional food, various herbal extracts, food additives, food additives, or natural carbohydrates may be added as additional ingredients.

The food supplement additive is added to prepare a health functional food of each formulation and can be appropriately selected and used by those skilled in the art. Flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, colorants and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents , Stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, etc., but the type is not limited by the examples.

At this time, the content of the composition contained in the food is not particularly limited, but may be included in an amount of 0.01 to 100% by weight, more preferably 1 to 80% by weight relative to the total weight of the food composition.

Specifically, the composition of the composition is not limited to this, but may be 1 to 1000 μM, specifically 1 to 800 μM, more specifically 1 to 600 μM, and even more specifically 1 to 500 μM.

When the food is a beverage, it may be included in a ratio of 1 to 30 g, preferably 3 to 20 g, based on 100 ml. In addition, the composition may include additional ingredients that are commonly used in food compositions to improve odor, taste, and vision. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantotenic acid, and the like. In addition, minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), and copper (Cu) may be included. In addition, amino acids such as lysine, tryptophan, cysteine, and valine may be included. In addition, preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), fungicides (bleached and highly bleached, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene ( BHT), etc., colorants (such as tar pigment), colorants (sodium nitrite, sodium nitrite, etc.), bleach (sodium sulfite), seasonings (such as MSG sodium glutamate), sweeteners (dulcine, cyclate, saccharin, sodium, etc.) , Food additives such as flavoring agents (vanillin, lactones, etc.), expanding agents (alum, D-potassium hydrogenate, etc.), strengthening agents, emulsifying agents, thickeners (foaming agents), coating agents, gum starting agents, foam inhibitors, solvents, improvers, etc. ) Can be added. The additive is selected according to the type of food and used in an appropriate amount.

Another aspect of the present invention provides a food composition comprising a composition for preventing browning of food comprising the macrorin of Formula 1 as an active ingredient.

The food composition of the present invention can be prepared by a method conventionally used in the art, and may be prepared by adding raw materials and ingredients conventionally added in the art. In addition, unlike general medicines, it has the advantage that there are no side effects that can occur when taking medicines for a long time using food as a raw material, and it can be excellent in portability.

The food composition is not limited thereto, but may include the food browning preventing composition in an amount of 0.001 to 25% by weight, specifically 0.01 to 20% by weight, and more specifically 0.01 to 10% by weight based on the total weight.

Another aspect of the present invention provides a food browning prevention method comprising the step of treating a food browning-preventing composition comprising the macrorin of Formula 1 as an active ingredient in food.

The "macroline" and "browning" are as described above.

Specifically, the method for preventing food browning of the present invention may include the step of treating the composition in a food-effective amount in a food that may or may have food browning.

The treatment may be any method that is conventionally used in the art as long as it can prevent browning of food using the composition, but may be specifically immersion treatment or spray treatment.

The composition comprising the macrorin of the present invention as an active ingredient can inhibit browning of fruits or vegetables that are easy to brown for a long time, and when added directly, has an antioxidant activity enhancing effect. In particular, since it has not only excellent anti-browning effect but also excellent anti-oxidation effect than vitamin C and arbutin, which were used as controls, the composition containing macrorin as an active ingredient suppresses browning and increases the antioxidant capacity of foods, thereby improving product quality as well as quality. Can be improved.

1 is a graph confirming the effect of inhibiting browning of potatoes over time using the method according to the embodiment.
2A, 2B, and 2C are graphs measuring the effect of macurine on long-term (5 weeks) shelf life of potatoes with a colorimeter.
3 is a graph showing absorbance of the effect on the activity of polyphenol oxidase (PPO) according to concentration when macurine is added to potatoes.
4A, 4B, 4C, and 4D are graphs showing the free radical scavenging activity of macrorin and the free radical scavenging ability in potato when macurine is added.
5 is a comparative experiment comparing the effect of inhibiting the activity of polyphenol oxidase by ethanol extract of mulberry and mulberry.
6A and 6B show graphs of antioxidant results and polyphenol oxidase activity inhibition results of substances that do not have an inhibitory effect on polyphenol oxidase activity, but do not have an antioxidant effect, and do not have an antioxidant effect. It is shown.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

Example 1: Analysis of the browning phenomenon of potatoes according to the amount of macurine added and the storage period

In order to determine whether the browning phenomenon of potatoes is inhibited by adding macrorin to the potato, an anti-browning analysis refers to a protocol described in a conventional article ( Food Chem . 2018 Jun 30; 252: 207-214). . First, the potato chips were crushed and centrifuged twice at 1000 rpm for 1 minute to harvest the potato supernatant. The harvested supernatant was treated with 5 μM to 500 μM of macrorin to observe changes in color over time.

As a result, as can be seen in Figure 1, polyphenol oxidase inhibitors and arbutin known to be capable of inhibiting browning (Arbutin) and vitamin C in a concentration-dependent manner was more excellent inhibitory effect of browning, especially arbutin And vitamin C had an unstable inhibitory effect over time, whereas macrorin showed stable anti-browning effect from 4 hours to 84 hours. Therefore, as shown in FIG. 1, it is determined that macrorin is an anti-browning agent and can be used as a food additive that inhibits browning (FIG. 1).

Example 2: Anti-browning effect of potato for a long time according to the amount of macurine

After a long period of time (5 weeks) by adding macrorin to the potato, the effect of preventing the browning of the potato was observed, and a colorimeter described in a conventional paper ( Food chemistry 201 (2016) 315-319) was used. As a specific experimental method, chromaticity was measured using a color difference meter (CR-300 series, Minolta Co.) to measure the degree of browning of each sample. At this time, the L value represents brightness, the a value represents redness, and the b value represents yellowness. It is known that L and a values are involved in browning of food.

As a result of the experiment, according to FIG. 2A, when 5 weeks elapsed after treatment with macurin, it was confirmed that the concentration-dependently high L value was compared to the control group, and the effect was superior to that of the arbutin treatment group, which is a positive control group.

According to Fig. 2b, macurine decreased the a value (redness) of potatoes in a concentration-dependent manner, but arbutin did not affect it. On the other hand, there was no difference in yellowness (Fig. 2c) in all groups. Therefore, through this result, it can be confirmed that when the potato is treated with macurine, a long-term browning-preventing effect can be obtained (FIG. 2).

Example 3: Inhibitory effect of polyphenol oxidase activity

By analyzing the inhibitory effect of polyphenol oxidase activity of macurine, the browning effect was specifically identified.

To confirm the effect of inhibiting polyphenol oxidase activity when macurine was added to potatoes, a protocol for measuring polyphenol oxidase activity described in a conventional paper ( Food Chemistry . 227 (2017) 102-110) was referred to. As a specific experimental method, 10 µl of DMSO (control), positive control vitamin C, arbutin, and macrorin were respectively treated in a 96-well microplate, 1 mM L-tyrosine buffer was added, and 20 µl of potato soup was processed in a refrigerator. Absorbance (450 nm) was measured by time using a microplate reader (Molecular Devices, Spectramax i3) while incubating for 84 hours.

In order to correct the intrinsic color of the positive control group or macrorin, the blank group was treated with 10 μl of the positive control group or macrorin, followed by 190 μl treatment with 1 mM L-tyrosine buffer, and potato soup was not treated. Calculation of the polyphenol oxidase activity inhibitory effect was calculated by subtracting the absorbance of the blank group from the absorbance of the sample group, and then setting the DMSO (control) treatment group to 100, thereby inhibiting the polyphenol oxidase activity inhibition effect of the positive control group and the macrorin treatment group. %.

As can be seen in the experimental results, it was confirmed that the inhibitory effect of polyphenol oxidase activity, which is the cause of browning, is superior to that of the macrocontrol group, as can be seen in FIG. 3. (Figure 3).

Example 4: Effect of promoting food antioxidant activity of macrorin

The measurement of ROS scavenging effect of macrorin was performed using a 2 ', 7'-dichlorodihydrofluorescein diacetate (DCFDA) assay described in a conventional paper ( Chem Res Toxicol . 5: 227-231, 1992).

As a specific experimental method, esterase (600 U / ml) dissolved in tertiary distilled water and DCFDA (12.5 mM) dissolved in ethanol (99.9%) were stored as a storage solution at -20 ° C, and when used in the experiment, 6 U / ml The 2 ', 7'-dichlorodihydrofluorescein (DCFH) solution prepared by mixing esterase and 10 mM DCFDA was incubated at 22 ° C for 20 minutes, and then stored frozen until use.

Thereafter, the fat-soluble DCFDA is deacetylated to non-fluorescent DCFH due to esterase or oxidative hydrolysis, and DCFH is oxidized by free radicals to become DCF (2 ', 7'-dichlorofluorescein), which exhibits strong fluorescence. It was measured with a fluorescence photometer (GENios, TECAN) at (excitation) wavelength 485 nm and emission wavelength 535 nm.

SIN-1 (3-morpholinosydnonimine hydrochloride) 10 μM was used as the active oxygen generator, and the positive control group was a vitamin C-treated group having a strong free radical scavenging effect.

As a result of test tube experiments in which potatoes were not added, as shown in FIG. 4, ROS increased rapidly after treatment with SIN-1, which is an active oxygen generating source, but the treatment group treated with SIN-1 and macrorin was SIN-. Compared to the control group treated with only 1 (10 μM), free radical production was significantly suppressed in a concentration-dependent manner, and the inhibitory effect was higher than that of the positive control vitamin C. (Figure 4a).

In addition, in FIG. 4B, it was confirmed that a marked antioxidant effect was observed in the potato with the addition of macrorin in the concentration of macrorin (FIG. 4B).

However, when vitamin C, which is a positive control, was added to potatoes, the ROS increased to SIN-1 decreased, but the ROS scavenging effect was less than that of the same concentration of macrorin. This means that even if it is a substance known as a powerful antioxidant, the degree of its effect may be different when added to food.

ONOO rurin of ^mark-erasing effect is a conventional paper (Kooy et al, Free Radic Biol Med 16:.... 149-156, 1994) to refer to, ONOO ^- oxidation of dihydrorhodamine 123 (DHR123) mediated by the reaction It was measured by fluorescence spectroscopy. -Free light-DHR123 is ONOO ^- there is generated a Rhodamine123 noticeable fluorescence is oxidized by, ONOO ^- was used for SIN-1 (10μM) to induce, the positive control group was of strong ONOO ^- Vitamin C treated group with a scavenging effect Did.

As a result of the experiment, as shown in FIG. 4C, ONOO ^- was rapidly induced by SIN-1, but the treatment group treated with SIN-1 and macrorin produced ONOO ^- compared to the control group treated only with SIN-1. This was significantly suppressed, and in particular, it was confirmed that ONOO ^- production was significantly inhibited similarly to the vitamin C-treated group, which is a positive control (FIG. 4C). Both macrorin and vitamin C completely inhibited ONOO ^- production at a concentration of 1 μM (FIG. 4C).

In addition, in FIG. 4D, it was confirmed that a marked antioxidant effect was observed in the potato with the addition of macrorin (FIG. 4D).

However, the positive control vitamin C was excellent in ONOO ^- scavenging ability in the absence of potatoes at 1 μM concentration (test tube) (FIG. 4C), but when added to potatoes, it did not reduce the ONOO ^- production increased by SIN-1. . On the other hand, macrorin showed excellent ONOO ^- eliminating effect even when added to potatoes. This result also means that even when a substance known as a strong antioxidant is added to food, its effect may be different.

Therefore, it was confirmed that macrorin has excellent free radical scavenging ability, and especially when added to food, it was confirmed that the food antioxidant enhancing effect was excellent. Through this, macrorin is an antioxidant, improving product quality and productability. By doing so, it can be usefully used as a food additive.

Example 5: Comparative experiment on the inhibitory effect of polyphenol oxidase of macurine and mulberry ethanol extract

Measurement of the polyphenol oxidase activity was carried out in the same manner as in the method set forth in Example 3 mentioned above. In the previous paper, a comparative experiment was conducted with a mulberry branch or mulberry root bark ethanol extract, which is known to contain polyphenol oxidase inhibitory effect and contains macrorin.

As shown in Fig. 5, the mulberry root bark or mulberry branch extract (20 µg / ml) reduced polyphenol oxidase activity by 28% and 36%, respectively, but the same concentration of macrorin reduced polyphenol oxidase activity. It was reduced by about 65% (Fig. 5). This means that the effect of inhibiting polyphenol oxidase of macrorin is superior to that of mulberry extract.

Example 6: Examples of a substance having an effect of inhibiting polyphenol oxidase activity even if it has an antioxidant effect or a substance having an effect of inhibiting polyphenol oxidase activity even without an antioxidant effect.

The following experiment was conducted to prove that the compound having antioxidant effect does not necessarily inhibit the polyphenol oxidase.

First, in order to measure the oxidative stress induced by SIN-1 treatment, peroxynitrate was induced by treating 50 μM SIN-1 in a 96-well plate, and to observe the change in fluorescence by adding DHR123, the excitation wavelength was 485 nm and Measurement was performed by a microtubule fluorescence reader at an emission wavelength of 535 nm at 5 minute intervals for 30 minutes. In addition, 10 μM penicillamin was used as a positive control.

As a result of performing the above experiment, as shown in FIG. 6A, the single substance F41 had a stronger antioxidant effect than the penicillamin used as a positive control. However, as a result of polyphenol oxidase (PPO) activity, F41 showed little inhibitory effect on the activity of polyphenol oxidase.

Conversely, as shown in FIG. 6B, the single substance F48 exhibited a strong inhibitory effect on polyphenol oxidase activity as much as kojic acid, but no antioxidant effect.

In sum, this experiment does not necessarily have an effect of inhibiting polyphenol oxidase activity because it has an antioxidant effect, and since the macrorin of the present invention has not only an antioxidant effect but also a polyphenol oxidase inhibitory effect, it has excellent food browning prevention effect. By exerting, it will be possible to improve not only the quality of food, but also the marketability.

In this specification, contents that can be sufficiently recognized and inferred by those of ordinary skill in the technical field of the present invention are omitted from the detailed description, and in addition to the specific examples described in the present specification, the technical idea or essential configuration of the present invention is changed. Various modifications are possible within a range that does not. Therefore, the present invention can be implemented in a different manner from the one specifically described and illustrated in the present specification, which can be understood by those of ordinary skill in the art.

Claims (8)

A composition for preventing food browning, comprising the following formula (maclurin) of formula 1 as an active ingredient.

[Formula 1]

The composition of claim 1, wherein the composition inhibits polyphenol oxidase.
The composition of claim 1, wherein the composition is added to food to enhance the antioxidant capacity of the food.
The composition of claim 1, wherein the composition has a macurine content of 1 to 500 μM.
A food composition comprising the composition for preventing food browning according to any one of claims 1 to 4.
The food composition of claim 5, wherein the composition for preventing food browning is contained in an amount of 0.01 to 10% by weight based on the total weight of the food composition.
A method of preventing food browning, comprising the step of treating the composition of claim 1 with food.
The method according to claim 7, wherein the treatment is spray treatment or immersion treatment.

Images