KR102150843B1 - Naturally occurring essential oil composition having antifungal activities against xerophilic fungi - Google Patents

Abstract

The present invention relates to an antifungal composition comprising natural essential oils, and the composition according to the present invention can effectively inhibit the growth of dry-resistant molds, as well as minimize the amount of essential oils used individually, thereby reducing the organoleptic properties of food when applied to food. Changes can be minimized, and cost savings can also be obtained by reducing the use of individual refineries.

Description

Naturally occurring essential oil composition having antifungal activities against xerophilic fungi for controlling dry mold

The present invention relates to a natural essential oil composition showing antifungal activity against dry mold, a food packaging material containing the same, a food packaging method, and a method for preventing or inhibiting the growth of dry mold resistant mold.

Penicillium corilophyllum Corruption in the 0.60 to 0.85 of the food): corylophilum) and Euro tium Les pence (a _w (water activity, Water Activity Eurotium naegeonseong fungi such as repens) is a relatively Microbiological Safety of Food is considered that the intermediate water (intermediate-moisture food) It is known to cause (Non-Patent Documents 1 to 3). Not only can P. corylophilum cause food spoilage, but it has also been reported that it produces citrinin and mycotoxin as a secondary product of metabolic processes (Non-Patent Document 4). In addition, since Penicillium species and Eurotium species are known to exist frequently in the environment, cross-contamination with food can easily occur.

Dry-resistant molds are generally resistant to various external stresses such as low pH, osmotic pressure, heat, and high salt concentration as well as in a dry environment, so they survive in food and can cause food poisoning in people who consume it. In order to control the growth of these dry-resistant molds, a method of spraying ethanol or the like that can be added to food on the food surface or adding synthetic chemical preservatives has been attempted, but there is no significant effect.

As the health-oriented demands of consumers have recently increased, there is an increasing tendency to prefer natural antibacterial agents such as essential oils rather than chemical synthetic substances as antibacterial agents. Essential oils are highly volatile liquids obtained by extraction from plants and are known to have antibacterial activity against various microorganisms. Essential oils having antibacterial activity against these microorganisms can be used in a liquid or gaseous state. Essential oil can be used as an antibacterial packaging material for food, and has the advantage of high permeability to microorganisms contaminated in the crevices of food surfaces or food contact surfaces. Is increasing. However, the biggest limitation in applying natural essential oils to foods is that natural essential oils generally have a strong flavor, so when used in foods, it is highly likely to affect the organoleptic properties and physical properties of the food. In addition, when one type of essential oil is used alone, there is a disadvantage in that it is expensive because a large amount is required in order to exhibit the antibacterial effect.

Therefore, in order to treat natural essential oil in food in order to control dry mold resistance, there is a need for a method capable of minimizing the amount of essential oil to be processed in food or the amount of essential oil remaining in food after treatment.

Leistner, L., 1992. Food preservation by combined methods. Food Research International, 25(2), 151-158. Suhr, K. I., & Nielsen, P. V., 2003. Antifungal activity of essential oils evaluated by two different application techniques against rye bread spoilage fungi. Journal of Applied Microbiology, 94(4), 665-674. Pitt, J. I., & Hocking, A. D., 2009. The ecology of fungal food spoilage. In Fungi and food spoilage (pp. 3-9). Springer, Boston, MA. Dos Santos, C. M. C., Costa, G. L. D., & Figueroa-Villar, J. D., 2012. Identification of citrinin as the defense metabolite of Penicillium corylophilum stressed with the antagonist fungus Beauveria bassiana. Nat. Prod. Res. 26, 2316-2322.

It is an object of the present invention to provide an antifungal composition capable of minimizing changes in organoleptic properties and cost of food by reducing the amount of individual essential oils while maintaining the antifungal effect of essential oils against dry mold.

The inventors of the present invention confirmed that the antibacterial synergy can be maximized by searching for the combination and ratio of natural essential oils that cause antibacterial synergy while conducting research on natural products for essential oil compositions that exhibit antifungal activity against dry mold. Was completed.

The present invention prevents or inhibits the growth of dry-resistant mold in food comprising the step of adding an antifungal composition against dry-resistant mold containing natural essential oil as an active ingredient, a food packaging material containing the antifungal composition, and the antifungal composition to the food packaging material It provides a food packaging method for preventing or inhibiting the growth of dry-resistant mold in food comprising the step of treating the antifungal composition to the food.

In the following examples, it was confirmed that the antifungal activity of the composition according to the ratio of natural essential oils against dry-resistant fungi was confirmed, and it was confirmed that the amount of essential oils individually used from the antifungal synergy by the combination of natural essential oils can be minimized. . Therefore, natural essential oils combined in a certain ratio can be used as an active ingredient of an antifungal composition against dry-resistant mold.

Hereinafter, the configuration of the present invention will be described in detail.

The present invention is cinnamon bark, citronella, May chang, Garlic, thyme linalool, oregano, thyme thymol, and spia It provides an antifungal composition against dry-resistant fungi comprising two or more essential oils selected from the group consisting of mint (Spearmint) as an active ingredient.

In the present invention, "essential oil" refers to an oily extract obtained by condensing a fragrance component from cinnamon peel, citronella, meichang, garlic, thyme linalul, oregano, thyme thymol or spearmint. For the production of the essential oil, a steam distillation method, a solvent extraction method using a volatile solvent (benzene, hexane, etc.), a leaching method (a method of collecting the leaching sap by wounding an extraction target), a compression method, a supercritical extraction method, etc. can be used. It is not limited thereto, and may be prepared using a method known in the art. In addition, the essential oil may use a commercially available product. The essential oil can be applied as a liquid or gas.

In one embodiment, the essential oil may be applied in a gaseous state.

In the present invention, the antifungal essential oil composition exhibits antifungal activity against dry-resistant mold.

In the present invention, dry-resistant mold refers to a mold that can grow even in relatively dry conditions, and refers to a mold that can grow even in a water activity of 0.60-0.85.

In one embodiment, the dry-resistant fungus may be Penicillium corylophilum or Eurotium repens .

The Penicillium corilofilum and Eurotium repens are fungi belonging to blue mold and millet mold, respectively, and may cause food corruption in low-moisture foods or medium-moisture foods. In addition, Penicillium corilofilum can cause food poisoning in people who consume it by producing mycotoxins.

Therefore, the composition according to the present invention can prevent food poisoning and deterioration of food quality by inhibiting the growth of the fungi.

In the present invention, the antifungal composition may include a combination of two or more essential oils selected from the group consisting of cinnamon peel, citronella, mei chang, garlic, thyme linal, oregano, thyme thymol, and spearmint.

In the present invention, the essential oil of the composition may include two essential oils combined in a concentration ratio of 1:0.1 to 1:32. For example, essential oil A and essential oil B have a concentration ratio of 1:0.1 to 1:32, 1:0.25 to 1:16, 1:0.25 to 1:8, 1:0.5 to 1:16, 1:0.5 to 1:8, It may be included in a concentration ratio of 1:0.5 to 1:4, 1:1 to 1:3, 1:2 to 1:4, 1:2 to 1:16, 1:4 to 1:8 and combined.

In one embodiment, the present invention is cinnamon peel essential oil; And

It may be an antifungal composition comprising, as an active ingredient, essential oil selected from the group consisting of citronella, oregano, May Chang, thyme linal and garlic.

In the combination, cinnamon peel essential oil; And the essential oil selected from the group consisting of citronella, oregano, May Chang, thyme linal and garlic may be combined in a concentration ratio of 1:0.01 to 1:32. For example, the combination essential oil may be included in a concentration ratio of 1:0.05 to 1:25, 1:0.0625 to 1:20, and 1:0.25 to 1:16.

In another embodiment, the present invention provides citronella essential oil; And

It may be an antifungal composition comprising, as an active ingredient, essential oil selected from the group consisting of garlic, May Chang, oregano, thyme thymol, spearmint, and thyme linalul.

In the above combination, Citronella essential oil; And the essential oil selected from the group consisting of garlic, May Chang, oregano, thyme thymol, spearmint, and thyme linalul may be combined in a concentration ratio of 1:0.01 to 1:32. For example, the combination essential oil may be included in a concentration ratio of 1:0.05 to 1:16, 1:0.1 to 1:8, and 1:0.125 to 1:1.

In another embodiment, the present invention is Mei Chang essential oil; And

It may be an antifungal composition comprising an essential oil selected from the group consisting of garlic, oregano, and thyme linal as an active ingredient.

In the above combination, Mei Chang essential oil; And the essential oil selected from the group consisting of garlic, oregano, and thyme linal may be combined in a concentration ratio of 1:0.01 to 1:32. For example, the combination essential oil may be included in a concentration ratio of 1:0.05 to 1:16, 1:0.1 to 1:8, and 1:0.5 to 1:1.

In another embodiment, the present invention is a garlic essential oil; And

It may be an antifungal composition comprising, as an active ingredient, essential oil selected from the group consisting of thyme linal, thymol, oregano, and spearmin.

In the above combination, garlic essential oil; And the essential oil selected from the group consisting of thyme linal, thymol, oregano, and spiramine may be combined in a concentration ratio of 1:0.01 to 1:32. For example, the combination essential oil may be included in a concentration ratio of 1:0.05 to 1:20, 1:0.1 to 1:16, and 1:2 to 1:8.

In another embodiment, the present invention provides spearmint essential oil; And

It may be an antifungal composition comprising oregano or thyme linalul essential oil as an active ingredient.

In the above combination, spearmint essential oil; And oregano or thyme linalul essential oil may be combined in a concentration ratio of 1:0.01 to 1:32. For example, the combination essential oil may be included in a concentration ratio of 1:0.05 to 1:16, 1:0.1 to 1:8, and 1:0.25 to 1:2.

In another embodiment, the present invention may be an antifungal composition comprising thyme linalul essential oil and thyme thymol essential oil as active ingredients.

In the above combination, thyme linarul essential oil and thyme thymol essential oil are combined in a concentration ratio of 1:0.01 to 1:8, such as 1:0.05 to 1:4, 1:0.1 to 1:2, and 1:0.5 to 1:1 It can be.

In the following examples, it was confirmed that the combination of cinnamon peel essential oil and citronella essential oil exhibited an antifungal synergistic effect against Penicillium corilofilum. Accordingly, as a result of confirming the antifungal effect by additionally combining cinnamon peel essential oil and citronella essential oil and Meichang essential oil each showing partial synergistic effect based on the above combination, the combination of the three essential oils was found to be penicillium corilofilum. It was confirmed that an antifungal synergistic effect appeared.

Accordingly, the present invention may include an antifungal composition comprising cinnamon peel essential oil, citronella essential oil, and Meichang essential oil as active ingredients.

In the above combination, the cinnamon peel essential oil, citronella essential oil and Mei Chang essential oil are in a concentration ratio of 1:0.5:0.5 to 1:4:4, such as 1:1:1 to 1:2:2, 1:2:1 To 1:2:2 may be combined in a concentration ratio.

In addition, according to the following examples, as a result of treating food by combining two types of essential oils selected from the group consisting of cinnamon peel, citronella, garlic, May Chang, thyme thymol, spearmint, thyme linalul and oregano, It was confirmed that there was a synergistic effect of antifungal partial against Tium repence.

In the present invention, the content of the essential oil with respect to the total weight of the composition may vary depending on the concentration of the essential oil to be combined and the object to which the composition of the present invention is applied.

Meanwhile, in the present specification, the term "included as an active ingredient" means containing an amount sufficient to achieve the antibacterial effect or activity of the essential oil as described above. In the present invention, the essential oil may be 0.0001 µl/mL or more, such as 0.001 µl/mL or more, or 0.004 µl/mL or more, but is not limited thereto.

In one embodiment of the present invention, the essential oil to be combined in the composition of the present invention is, for example, when cinnamon skin essential oil and citronella essential oil are combined, cinnamon skin essential oil is 0.0001 µl/mL or more, preferably 0.001 µl. /mL or more, more preferably 0.0015 µl/mL or more is contained, and Citronella essential oil contains 0.0001 µl/mL or more, preferably 0.001 µl/mL or more, and more preferably 0.0015 µl/mL or more.

In another embodiment of the present invention, when the combination of Mei Chang essential oil is further combined, the Mei Chang essential oil is 0.0001 μL/mL or more, preferably 0.001 μL/mL or more, and more preferably 0.0015 μL/mL or more. Can be included.

Since the essential oil is a natural product and does not have side effects on the human body even when administered in an excessive amount, the upper limit of the quantity of the essential oil contained in the composition of the present invention can be selected and implemented within an appropriate range by a person skilled in the art.

In addition, the present invention provides an antifungal food packaging material for inhibiting the growth of dry-resistant mold comprising the antifungal composition. In one embodiment, the dry-resistant fungus is Penicillium corilofilum ( Penicillium corylophilum ) or Eurotium repens .

According to the present invention, when the food packaging material is used, the amount of essential oil used is minimized compared to the amount of essential oil used in the conventional food packaging material, thereby minimizing the sensory effect on the food, and cost reduction.

In addition, the present invention provides a food packaging method for inhibiting the growth of dry-resistant mold in food, comprising the step of applying or coating the antifungal composition on a food packaging material.

In the present invention, the step of applying or coating the antifungal composition on a food packaging material includes cinnamon bark, citronella, May chang, Garlic, thyme linal (Thyme). linalool), oregano (Oregano), thyme thymol (Thyme thymol) and spearmint (Spearmint) by combining two or more essential oils selected from the group consisting of to prepare an antifungal composition may be further included.

In the above method, specific combinations of essential oils and antifungal compositions may be equally applied or applied mutatis mutandis to the above description.

In addition, the present invention provides a method for preventing the growth of dry-resistant mold in food, comprising the step of treating the antifungal composition to the food.

In the present invention, the dry-resistant mold may be Penicillium corylophilum or Eurotium repens .

In the present invention, the food may be a low water activity food or an intermediate-moisture food. The low moisture food and medium moisture foods refer to foods having a moisture content of about 1 to 20% naturally occurring or by a drying or dehydration process. Food groups belonging to the low moisture food and medium moisture food may be grains (whole grains and whole grains), seasonings, spices, dry herbs, powdered foods, coffee, breads, confectionery, dried fruits, dried vegetables, seeds, or jerky. It is not limited.

In the present specification,'water activity' is the ratio (Ps/Po) of the water vapor pressure (Po) of pure water to the water vapor pressure (Ps) represented by moisture in food at an arbitrary temperature (Ps/Po), growth of microorganisms, flavor of food It is related to chemical, biological, and physical reactions in food that change color and flavor.

The low moisture food and medium moisture food generally have a water activity of 0.60 to 0.85.

In the present invention, the step of treating the antifungal composition on food may be a step of spraying the composition on the food.

The antifungal composition of the present invention not only can effectively inhibit the growth of dry mold-resistant fungi, but also minimizes the amount of essential oils individually used due to the antifungal synergy that can be obtained by combining essential oils, thereby reducing the change in sensory properties of food when applied to food. It can be minimized, and cost savings due to the reduction in the use of individual refineries can also be obtained.

1 is a schematic diagram of a method of measuring the minimum inhibitory concentration (MIC) of a refinery gas for P. corylophilum .
Figure 2 is a schematic diagram of a checkerboard assay method of cinnamon peel essential oil and citronella essential oil for P. corylophilum .
3 is a schematic diagram of a sealed container for confirming the antibacterial activity of a combination of cinnamon peel essential oil, citronella essential oil, and Meichang essential oil for P. corylophilum inoculated on jerky.

Advantages and features of the present invention, and a method of achieving them will become apparent with reference to embodiments described below in detail. However, the present invention is not limited to the embodiments disclosed below, but will be implemented in a variety of different forms, only the present embodiments are intended to complete the disclosure of the present invention, and the general knowledge in the technical field to which the present invention pertains. It is provided to completely inform the scope of the invention to those who have it, and the invention is only defined by the scope of the claims.

[ Example One] Penicillium Corylophilum ( P. corylophilum )on Determination of the minimum inhibitory concentration of natural oil gas in Korea

In order to determine the minimum inhibitory concentration (MIC) of natural essential oil for P. corylophilum , a gas antibacterial activity measuring device and a gas antibacterial activity measuring method (Korean Patent No. 10-1406387) were used.

First, P. To prepare the extract of spore corylophilum, Dichloran 18% were incubated with P. corylophilum Glycerol agar (DG18) and the frequency divider 0.1% peptone water with 30% glycerol (0.1% PW30G) in 10 mL of the agar medium, After separating the colonies on the culture medium in which the fungi were cultured using a spreader, the suspension in which the colonies of P. corylophilum and 0.1% PW30G were mixed was filtered using sterilized gauze. The number of spores in the suspension was counted using a Neubauer counting chamber (hemocytometer) and a microscope. The spore suspension was diluted to a concentration of 10 ⁵ or 10 ⁶ spores/mL using 0.1% PW30G, and then the diluted spore suspension was mixed by 5 mL to prepare 10 mL of P. corylophilum spore extract to be used in the experiment.

DG18 was prepared and autoclaved. After adding 0.35 mL each of the liquid medium to the upper container of the antimicrobial activity measuring device, and hardening, 10 µl of P. corylophilum spore extract (ca. 10 ⁶ spores/mL) is dispensed to allow microorganisms to be adsorbed to the medium laminal flow biosafety hood (22). ±2°C) for 30 minutes. Natural essential oils were binary diluted step by step using dimethyl sulfoxide (DMSO). After dispensing 10 µl of the natural essential oil dilution into the lower container of the antimicrobial activity measuring device containing the paper disc (Advantec Toyo Kaisha. Ltd., Tokyo, Japan), the upper container and the lower container were quickly combined and sealed. The sealed containers were cultured in a 25° C. incubator for 120 hours, and then the presence or absence of colony formation on the medium was visually observed, and the minimum concentration of the essential oil gas in which colonies were not formed was set as the minimum inhibitory concentration (MIC). 1 shows a method of measuring the MIC of natural oil gas for P. corylophilum , and the measurement results are shown in Table 1.

essential oil Minimum inhibitory concentration (µl/ml) Scientific name Common name Area of use Allium sativum Garlic Root 0.0390 Cinnamomum zeylanicum Cinnamon bark Bark 0.1563 Litsea cubeba May chang Fruit 0.1563 Cymbopogon nardus Citronella Grass 0.3125 Thymus zygis CT thymol Thyme thymol Leaf 0.3125 Origanum vulgare Oregano Leaf 0.6250 Mentha spicate Spearmint Flowering herb 0.6250 Thymus zygis CT linalool Thyme linalool Flower 0.6250

As shown in Table 1, all of the garlic, cinnamon peel, May Chang, citronella, thyme thymol, oregano, spearmint and thyme linalul essential oil showed a low minimum inhibitory concentration (MIC) of 0.6250 μl/mL or less . It was confirmed that the antifungal effect against corylophilum was excellent.

[ Example 2] Cinnamon Shell essential oil gas and Citronella Refinery gas Combination Checking antifungal synergy

Currently, the experimental method for confirming antibacterial synergy by combining natural essential oils is not standardized, so a checkboard assay, which is a method of confirming antibacterial synergy by combining liquid antibacterial substances, was applied and used.

The antibacterial activity measuring device (vial type) was arranged in 4 rows by 4 columns. After adding 0.35 mL of DG18 agar (refer to Example 1) to the upper container of the antimicrobial activity measuring device and hardening for 30 minutes, 10 µl of P. corylophilum spore extract (10 ⁶ spores/mL) was dispensed and the microorganisms were adsorbed to the medium by laminar flow. It was maintained for 30 minutes in a biosafety hood (22±2℃). An 8 mm diameter paper disc (Advantec Toyo Kaisha. Ltd., Tokyo, Japan) divided in half was placed in the lower container of the antibacterial activity measuring device. The liquid refinery gases were sequentially diluted in binary using DMSO until they reached 1/8 of the minimum inhibitory concentration (MIC) of the refinery gas. 2.5 µl of essential oil A, binary diluted from MIC concentration to 1/8 MIC concentration, was dispensed in the horizontal direction (4 vials) of 16 vials, and binary diluted from MIC concentration to 1/8 MIC concentration in the vertical direction (4 vials). 2.5 μl of essential oil B was dispensed. Immediately after dispensing, the upper and lower containers were quickly joined and sealed. The sealed containers were cultured in a 25° C. incubator for 120 hours, and then the presence or absence of colonies on the medium was visually observed. When the colony of P. corylophilum was not formed, the fractional inhibitory concentration (FIC) and the fractional inhibitory concentration index (FICI) were calculated to confirm the occurrence of antifungal synergistic effects. At this time, if FICI is less than or equal to 0.5, there is a synergistic effect, if it is between 0.5 and 1, a partial synergistic effect, if it is between 1 and 4, it is independent, and if it is 4 or more, it is an antagonistic effect. It was judged that there was. The measurement method is shown in Fig. 2 and the measurement results are shown in Table 2.

[Equation]

Refinery A Refinery B Essential oil concentration (µl/mL) FIC FICI result Refinery A Refinery B Total refinery Refinery A Refinery B Cinnamon Cinnamon bark Citronella (Citronella) 0.0390 0.0781 0.1171 0.2500 0.2500 0.5000 Synergy Garlic Citronella 0.0195 0.0390 0.0585 0.5000 0.1250 0.6250 Partial synergy Garlic Thyme linalool 0.0195 0.0781 0.0976 0.5000 0.1250 0.6250 Partial synergy May chang Cinnamon bark 0.0195 0.0781 0.0976 0.1250 0.5000 0.6250 Partial synergy May chang Citronella 0.0195 0.1563 0.1758 0.1250 0.5000 0.6250 Partial synergy Citronella Oregano 0.1563 0.0781 0.2344 0.5000 0.1250 0.6250 Partial synergy Cinnamon bark Oregano 0.0195 0.3125 0.3320 0.1250 0.5000 0.6250 Partial synergy Garlic Thyme thymol 0.0195 0.0390 0.0585 0.5000 0.2500 0.7500 Partial synergy Garlic May chang 0.0195 0.0390 0.0585 0.5000 0.2500 0.7500 Partial synergy Garlic Oregano 0.0195 0.1563 0.1758 0.5000 0.2500 0.7500 Partial synergy May chang Oregano 0.0781 0.1563 0.2344 0.5000 0.2500 0.7500 Partial synergy Garlic Spearmint 0.0195 0.1563 0.1758 0.5000 0.2500 0.7500 Partial synergy Citronella Thyme thymol 0.1563 0.0390 0.1953 0.5000 0.2500 0.7500 Partial synergy Citronella Spearmint 0.1563 0.1563 0.3125 0.5000 0.2500 0.7500 Partial synergy Citronella Thyme linalool 0.1563 0.1563 0.3125 0.5000 0.2500 0.7500 Partial synergy Thyme linalool Spearmint 0.3125 0.1563 0.4688 0.5000 0.2500 0.7500 Partial synergy

As shown in Table 2, synergistic or partial synergistic effects were confirmed in the experimental group of various combinations of essential oils for P. corylophilum . Among them, it was found that when the cinnamon peel essential oil and citronella essential oil were combined in a concentration ratio of 1:2, the growth of P. corylophilum could be suppressed in a smaller amount than when each essential oil was treated alone. . As the FICI value was 0.5000 in the combination treatment group, it was confirmed that the combination of the two essential oil gases had an antifungal synergistic effect against P. corylophilum .

[ Example 3] Cinnamon Peel essential oil, Citronella Refinery and may Window refinery Combination Check antibacterial synergy

In Example 2, the antifungal synergistic effect against P. corylophilum was confirmed in the combination of cinnamon peel essential oil and citronella essential oil. Accordingly, the experiment was carried out by additionally combining the gaseous of Meichang essential oil, which exhibited a partial synergy effect (FICI = 0.6250) in common when combined with the cinnamon shell essential oil and citronella essential oil, respectively.

Antimicrobial activity measuring devices (64 vials) were arranged in 4 rows × 4 rows × 4 sets. Add 0.35 mL of DG18 agar to the upper container of the antimicrobial activity measuring device and harden for 30 minutes, and then dispense 10 µl of P. corylophilum spore extract (10 ⁶ spores/mL) to allow microorganisms to be adsorbed to the medium laminal flow biosafety hood (22±2) ℃) for 30 minutes. An 8 mm diameter paper disc (Advantec Toyo Kaisha, Ltd., Tokyo, Japan) was placed in the lower container of the antibacterial activity measuring device. The liquid refinery gases were sequentially binary diluted from MIC of the refinery gas to 1/8 of the MIC using DMSO. 1.7 µl of essential oil A, binary diluted from MIC concentration to 1/8 MIC concentration, was dispensed in the horizontal direction (4 vials) of 16 vials, and binary diluted from MIC concentration to 1/8 MIC concentration in the vertical direction (4 vials). 1.7 μl of essential oil B was dispensed. Thus, four sets of antimicrobial activity measuring devices consisting of 16 vials were prepared. Thereafter, 1.7 μl of essential oil C obtained by binary dilution from the MIC concentration to the 1/8 MIC concentration was dispensed into each experimental apparatus. Immediately after dispensing, the upper and lower containers were quickly joined and sealed. The sealed containers were incubated in an incubator at 25° C. for 120 hours, and then the presence or absence of colonies on the medium was visually observed, and the antifungal synergy effect of the combination of essential oils was calculated by calculating FIC and FICI according to the same method as in Example 2. Confirmed. The results are shown in Table 3 below.

Essential oil concentration (µl/mL) FIC FICI result Cinnamon skin Citronella may window Total refinery Cinnamon skin Citronella may window 0.0195 0.0390 0.0195 0.0781 0.1250 0.1250 0.1250 0.3750 Synergy 0.0195 0.0390 0.0390 0.0975 0.1250 0.1250 0.2500 0.5000 Synergy 0.0195 0.0390 0.0781 0.1366 0.1250 0.1250 0.5000 0.7500 Partial synergy

In Example 2, the concentrations of cinnamon skin essential oil and citronella essential oil, which exhibited antifungal synergistic effects against P. corylophilum , were fixed, and the concentration of the Meichang essential oil gas was different. As a result, cinnamon skin essential oil, citronella When treating essential oils and Meichang essential oils in a concentration ratio of 1:2:1 or 1:2:2, it was confirmed that the growth of P. corylophilum can be inhibited in a smaller amount than that of each essential oil alone. I did. As the FIC values in the combination treatment group were 0.3750 and 0.5000, respectively, it can be seen that the combination of the three oil gases has an antifungal synergistic effect.

[ Example 4] Confirmation of antifungal effects of essential oil gases in actual medium moisture food

In the previous example, cinnamon peel essential oil, citronella essential oil, and Meichang essential oil, which showed strong antibacterial activity against P. corylophilum , and the combination that showed the lowest FICI in the combination treatment (1:2:1 concentration ratio (volume ratio)) The antimicrobial activity of was measured for P. corylophilum inoculated on jerky.

A sealed container for the experiment was prepared using a circular polystyrene container. 3 is a schematic diagram of a sealed container manufactured to treat essential oil gas on the jerky surface. A small Petri dish lid was placed to place the jerky coupons inside the sealed container. The upper part of the essential oil gas used to measure the MIC was turned over and placed next to the lid of a small Petri dish, and a sterilized 8 mm paper disc was placed inside the upper part of the oil gas frame. In order to maintain the internal relative humidity of the sealed container at 93%, 20 mL of saturated potassium nitrate was placed on the bottom of the sealed container at least 12 hours before the experiment, and then stored at 25°C.

After sterilizing the jerky coupon cut into 2 cm × 4 cm at room temperature (22±2°C) with ultraviolet rays, 100 µl of P. corylophilum spore mixture (ca. 10 ⁵ spores/mL) was inoculated at 10 points on the jerky coupon. spot inoculation) and dried for 1 hour in a laminar flow biosafety hood (22±2°C). The dried jerky coupons were placed on the lid of a small Petri dish inside the sealed container, and liquid essential oil was dispensed onto a paper disc. The amount of liquid essential oil dispensed was such that the final concentration of the essential oil gas when all of the liquid was vaporized in the sealed container was 0.0097, 0.0195, 0.0390 or 0.0781. In the case of the essential oil gas combination treatment, the concentration of cinnamon shell essential oil, citronella essential oil, and Meichang essential oil liquid was dispensed on a paper disc so that the concentration ratio was 1:2:1. After dispensing the liquid essential oil, the sealed container was sealed using parafilm and incubated at 25° C. for 120 hours.

Jerky coupons in which P. corylophilum was cultured were placed in a Whirl-pak bag containing 10 mL of 0.1% PW30G, and then pummeled for 1 minute using a homogenization device (Stomacher). After diluting the mixture of jerky and 0.1% PW30G by decimal using 0.1% PW30G, it was plated on DG18 agar medium. DG18 agar medium was incubated at 25° C. for 48 hours, and then colonies (dark green colonies) estimated to be P. corylophilum were counted and shown in Table 4 below. The theoretical limit of detection of the plate smear was 0.1 log CFU/cm ² .

essential oil P. corylophilum Number of objects (log CFU / cm ² ) Initial value Essential oil concentration (µl/mL) 0 0.0097 0.0195 0.039 0.0781 Cinnamon skin A2.9±0.1c A5.2±0.3a B4.4±0.3b A4.5±0.3b AB3.2± 0.4c B3.1±0.3c Citronella A2.9±0.1e A5.2±0.3a A4.8±0.2b B4.0±0.3c A3.7±0.5cd A3.6±0.4d may window A2.9±0.1d A5.2±0.3a A5.2±0.3a B4.3 ±0.2b AB3.5±0.3c B2.9± 0.3d Cinnamon skin, Citronella And Mei Chang combination A2.9±0.1b A5.2±0.3a A4.9±0.1a C3.2± 0.0b B3.0±0.4b B3.1±0.3b

In Table 4, capital letters in the same column mean no significant difference ( P≤0.05 ), and lowercase letters mean no significant difference in the same row ( P≤0.05 ).

As shown in Table 4, the initial population of P. corylophilum inoculated on the jerky surface was ca. It was 2.9 log CFU/cm ^2, and the population was ca. It increased to 5.2 log CFU/cm ² . The minimum concentration of each essential oil gas that did not show a significant difference from the initial population of P. corylophilum was cinnamon peel essential oil (0.0390 µl/mL), citronella essential oil (>0.0781 µl/mL), and Meichang essential oil (0.0781 µl/mL). mL) and three essential oil gases (0.0195 μl/mL). At a concentration of 0.0195 µl/mL, the number of P. corylophilum when treated with a combination of cinnamon husk essential oil, citronella essential oil, and Meichang essential oil gas (concentration ratio of 1:2:1 (volume ratio)) was determined as essential oil gas. Compared to the number of individuals in a single treatment, it was significantly lower ( P≤0.05 ). This result indicates that the combination treatment of the three essential oil gases can inhibit the growth of P. corylophilum present in the jerky at a lower concentration than the case of treatment with a single essential oil gas.

[ Example 5] Eurotium Refence( Eurotium repens )on Determination of the minimum inhibitory concentration of natural oil gas in Korea

To determine the minimum inhibitory concentration (MIC) of the natural essential oil for E. repens, the experiment was performed in the same manner as in Example 1, except that instead of the E. repens P. corylophilum the first embodiment. The results are shown in Table 5 below.

essential oil Minimum inhibitory concentration ( MIC )(Μl/mL) Scientific name Common name Use part Allium sativum Garlic Root 0.0390 Cinnamomum zeylanicum Cinnamon bark Bark 0.1563 Cymbopogon nardus Citronella Grass 0.1563 Litsea cubeba May chang Fruit 0.1563 Origanum vulgare Oregano Leaf 0.1563 Mentha spicate Spearmint Flowering herb 0.3125 Thymus zygis CT linalool Thyme linalool Flower 0.3125 Thymus zygis CT thymol Thyme thymol Leaf 0.3125

As shown in Table 5, all of garlic, cinnamon peel, May Chang, citronella, thyme thymol, oregano, spearmint and thyme linalul essential oil showed a low minimum inhibitory concentration (MIC) of 0.3125 μl/mL or less . It was confirmed that the antifungal effect against repens was excellent.

[ Example 6] Eurotium Refence( Eurotium repens )on For refinery gas Combination Checking antifungal synergy effect

In order to confirm the antifungal synergy against Eurotium repense by combining natural essential oils, an experiment was performed in the same manner as in Example 2. The results are shown in Table 6.

Refinery A Refinery B Essential oil concentration (µl/mL) FIC FICI result Refinery A Refinery B Total refinery Refinery A Refinery B Cinnamon bark May chang 0.0781 0.0195 0.0976 0.5000 0.1250 0.6250 part
Synergy Cinnamon bark Garlic 0.0781 0.0049 0.0830 0.5000 0.1250 0.6250 part
Synergy Cinnamon bark Citronella 0.0781 0.0195 0.0976 0.5000 0.1250 0.6250 Partial synergy Garlic Thyme thymol 0.0049 0.1563 0.1612 0.1250 0.5000 0.6250 Partial synergy Garlic May chang 0.0195 0.0195 0.0390 0.5000 0.1250 0.6250 Partial synergy Spearmint Thyme linalool 0.0390 0.1563 0.1953 0.1250 0.5000 0.6250 Partial synergy Cinnamon bark Thyme linalool 0.0781 0.0781 0.1563 0.5000 0.2500 0.7500 Partial synergy Garlic Spearmint 0.0195 0.0781 0.0976 0.5000 0.2500 0.7500 Partial synergy Garlic Thyme linalool 0.0097 0.1563 0.1660 0.2500 0.5000 0.7500 Partial synergy Garlic Oregano 0.0195 0.0390 0.0585 0.5000 0.2500 0.7500 Partial synergy May chang Thyme linalool 0.0781 0.0781 0.1563 0.5000 0.2500 0.7500 Partial synergy Oregano Spearmint 0.0390 0.1563 0.1953 0.2500 0.5000 0.7500 Partial synergy Thyme linalool Thyme thymol 0.1563 0.0781 0.2344 0.5000 0.2500 0.7500 Partial synergy

As shown in Table 6, partial synergistic effects were confirmed in the experimental group of various combinations of refinery gases for E. repens . Among them, a combination of cinnamon peel and Meichang essential oil, a combination of cinnamon peel and garlic essential oil, a combination of cinnamon peel and citronella essential oil, a combination of garlic and thyme thymol essential oil, a combination of garlic and Meichang essential oil, and spearmint and thyme Lina The combination of the rule essential oil showed a FICI value of 0.6250, confirming that there was a partial antifungal synergy effect.

Claims (15)

Cinnamon bark (Cinnamon bark) essential oil and citronella (Citronella) an antifungal composition for penicillium corylophilum ( Penicillium corylophilum ) containing essential oil as an active ingredient. The method of claim 1,
The cinnamon peel essential oil and citronella essential oil is an antifungal composition contained in a volume ratio of 1:0.5 to 1:4. The method of claim 1,
The composition is an antifungal composition further comprising a May chang essential oil as an active ingredient. The method of claim 3,
Cinnamon peel essential oil, citronella essential oil, and Meichang essential oil are antifungal compositions contained in a volume ratio of 1:0.5:0.5 to 1:4:4. delete delete delete delete delete delete delete An antifungal food packaging material for Penicillium corylophilum comprising the antifungal composition according to any one of claims 1 to 4. Food packaging for preventing or inhibiting the growth of Penicillium corylophilum in food, comprising the step of applying or coating the antifungal composition according to any one of claims 1 to 4 on food packaging Way. In a food product comprising the step of treating the antifungal composition according to any one of claims 1 to 4 to the food, Penicillium corylophilum ) to prevent or inhibit the growth. The method of claim 14,
The food is a method for preventing or inhibiting the growth of Penicillium corilofilum in a food that is a low moisture food or a medium moisture food.

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