WO2012015242A2 - Nouvelle composition antimicrobienne, et son procédé de préparation - Google Patents

Nouvelle composition antimicrobienne, et son procédé de préparation Download PDF

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WO2012015242A2
WO2012015242A2 PCT/KR2011/005541 KR2011005541W WO2012015242A2 WO 2012015242 A2 WO2012015242 A2 WO 2012015242A2 KR 2011005541 W KR2011005541 W KR 2011005541W WO 2012015242 A2 WO2012015242 A2 WO 2012015242A2
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extract
wheat
wheat bran
wheat germ
antimicrobial composition
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PCT/KR2011/005541
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Korean (ko)
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WO2012015242A3 (fr
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권영인
장해동
이학성
하경수
조성훈
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한남대학교 산학협력단
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3472—Compounds of undetermined constitution obtained from animals or plants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents

Definitions

  • the present invention relates to a novel antimicrobial composition and a method for producing the same, to an antimicrobial composition containing wheat germ extract or wheat bran extract having antimicrobial efficacy and a method for extracting antimicrobial components from wheat germ or wheat bran. More specifically, the present invention is an antimicrobial composition containing wheat germ extract or wheat bran extract as an active ingredient and an antimicrobial component from wheat germ or wheat bran as an active ingredient, which can replace preservatives or antibiotics added to food, cosmetics or animal feed, etc. It is about how to extract.
  • the present invention is a new natural antimicrobial material for preventing and treating pathogenic diseases caused by foods and preventing increased resistance to antibiotics, and is applicable to various fields such as food, beverages, cosmetics and food packaging.
  • Preservatives that can safely store food for a long time according to mass production and distribution of food, etc. are commonly used as additives in food or cosmetics. If preservatives are not used, food-borne pathogens may multiply in food or cosmetics and cause fatal harm to those who consume it or apply it to the skin. Therefore, preservatives that have been proven safe for humans have been widely used in foods and cosmetics. However, these preservatives have been reported as a factor that causes allergic diseases, atopic dermatitis, or even cancer by disturbing human immune function according to recent studies.
  • antibiotics and antimicrobials have recently reduced the mix of antibiotics and antimicrobials in formulated feeds to produce safe livestock products, while adding common antibiotics for both humans and animals to feed for livestock and farmed fish. Since 2012, measures have been banned.
  • Representative common antibiotics include penicillin, tetracycline-based antibiotics, dacitracin zinc, colistin sulfate, neomycin sulfate, and neomycin hydrochloride, and these acquired antibiotics will be banned in the future in the manufacture of animal feed.
  • Wheat germ is a nutritious embryo of malt (wheat kernel) that is removed from the process of processing whole wheat into flour, which accounts for about 2-3% of the whole wheat grain.
  • Wheat germ which is a seed of wheat, is known to be rich in vitamin E (tocopherol), which is known as an antioxidant vitamin to date, and recent studies have shown that natural substances extracted from wheat germ have the effect of restoring the immune function of the human body.
  • vitamin E tocopherol
  • Avemar a water-soluble extract obtained by fermenting wheat germ with yeast at 30 ° C for 18 hours, has been reported to have an effect of halving fever or other complications caused by the use of anticancer products. In the case of chemo side effects have been reported to reduce by more than 50%.
  • Korean Patent No. 10-0376538 discloses a hair shampoo composition for strengthening the hair internal fiber containing wheat germ protein as an additive, but as part of the shampoo composition It can be said that it stays at the level which utilizes the efficacy of the tocophenol which is the known component of wheat germ as mentioned above. Republic of Korea Patent No.
  • 10-0970544 discloses a skin external preparation with excellent whitening and anti-aging effect, it discloses the extract of the main ingredient Sisters and plant extracts and a variety of plants added to it, as an additional additive extract wheat germ extract It is mentioned.
  • the efficacy of the external skin preparation of the Republic of Korea Patent No. 10-0970544 mainly relates to the sister plant extract and wheat germ extract is referred to simply as an additional additive extract, and there is no disclosure about the exact efficacy of the wheat germ extract.
  • 10-2010-0038621 discloses a cosmetic composition for improving atopic dermatitis containing wheat germ oil and ceramide in the form of nano liposomes, strengthening the skin moisturizing power by reducing the amount of water loss by using wheat germ oil It only discloses the efficacy of improving atopic dermatitis, not only discloses or implies specific immune-improving efficacy, but does not suggest any antibacterial effect.
  • wheat bran also known as "bran" is the remainder of the separation of flour and embryos from milled wheat, most of which are seed blood. In the milling process, three kinds of sheaths, sophies, and flakes come out but are mixed and shipped from the factory for feed. Wheat bran has a lot of fiber, but has a lot of protein, fat, minerals, etc., attracting attention as a feed.
  • 10-1994-0013355 describes a method for preparing noodles with soft, chewy and savory flavor by adding wheat flour processed flour to wheat flour, but this also provides ingredients that give elasticity to noodles. It is only about improving the texture of the noodles using the physiological activity of wheat bran has not been disclosed at all.
  • wheat germ extract or wheat bran extract which is a natural material
  • wheat germ extract or wheat bran extract has antimicrobial effects against food-borne pathogens. There was no study or report on this.
  • the present inventors are interested in the research on wheat germ extract or wheat bran extract, which is a natural material, and the wheat germ extract or wheat bran extract extracted by the extraction method developed by the present inventors has excellent antimicrobial efficacy against major food pathogens. It was confirmed that there is an excellent antiseptic effect also in cosmetics and came to complete the present invention.
  • the present invention provides a novel antimicrobial composition and a method for producing the same.
  • the present invention is an antimicrobial composition containing wheat germ extract or wheat bran extract as an active ingredient and an antimicrobial component from wheat germ or wheat bran as an active ingredient, which can replace preservatives or antibiotics added to food, cosmetics or animal feed, etc.
  • the purpose is to provide a method for extracting.
  • the present invention is a new natural antimicrobial material for preventing and treating pathogenic diseases caused by foods and preventing increased resistance to antibiotics, and is applicable to various fields such as food, beverages, cosmetics and food packaging. It is an object of the present invention to provide an antimicrobial composition containing wheat germ extract or wheat bran extract having possible antimicrobial efficacy and a method for extracting the antimicrobial component from wheat germ or wheat bran.
  • the present invention provides an antimicrobial composition comprising wheat germ extract or wheat bran extract having an antimicrobial effect as an active ingredient.
  • the wheat germ extract or wheat bran extract is characterized in that extracted with at least one solvent selected from the group consisting of chloroform, methylene chloride, ethyl acetate and diethylene glycol monoethyl ether.
  • the wheat germ extract or wheat bran extract is obtained by further filtration after extraction with the solvent.
  • the wheat germ extract or wheat bran extract is characterized in that extracted by the supercritical fluid extraction method.
  • the supercritical fluid used in the supercritical fluid extraction method may be carbon dioxide and the cosolvent may be ethyl alcohol.
  • the wheat germ extract or wheat bran extract is obtained by further filtration after mixing with the co-solvent.
  • the wheat germ extract or wheat bran extract is characterized in that it is produced in a dry powder form through a drying process.
  • the antimicrobial composition of the present invention may include 0.01 wt% to 100 wt% of the wheat germ extract or wheat bran extract based on the total weight of the composition.
  • the antimicrobial efficacy composition of the present invention may be used as an antimicrobial substance to replace an antimicrobial agent added to an animal feed or an antimicrobial agent to replace a preservative added to food, beverage or cosmetics.
  • step (b) re-dissolving the wheat germ extract or wheat bran extract obtained in step (a) in the solvent and then filtering.
  • the method of preparing an antimicrobial composition of one embodiment of the present invention further comprises the step of evaporating the wheat germ extract or wheat bran extract in a vacuum evaporator to a maximum temperature of approximately 40 ° C. in the step (a).
  • the step (b) comprises the wheat germ extract or wheat bran extract obtained in step (a), or an evaporated concentrate of wheat germ extract or a wheat bran extract. After re-dissolving in the solvent, it is characterized in that the filtration through a PTFE (polytetrafluoroethylene) filter.
  • PTFE polytetrafluoroethylene
  • step (b) mixing the wheat germ extract or wheat bran extract obtained in step (a) with the co-solvent and filtering the mixture.
  • the step (b) is a wheat germ extract or wheat bran extract obtained in the step (a) after mixing with the co-solvent through a PTFE (polytetrafluoroethylene) filter It is characterized in that the filtration treatment.
  • PTFE polytetrafluoroethylene
  • the antimicrobial composition containing wheat germ extract or wheat bran extract having the antimicrobial efficacy of the present invention may replace preservatives or antibiotics added to food, cosmetics or animal feed.
  • wheat germ and wheat bran can be obtained in large quantities inexpensively and easily as a by-product of the flour production process, so the antimicrobial composition containing wheat germ extract or wheat bran extract of the present invention can be widely used in food, beverages, cosmetics, etc.
  • the antimicrobial composition containing wheat germ extract or wheat bran extract of the present invention can be used as a novel natural antimicrobial material for preventing and treating pathogenic diseases such as food-borne media and preventing increased resistance to antibiotics, As well as animal feed, there is an advantage that can be applied to various fields such as food, beverage, cosmetics and food packaging.
  • FIG. 1 is a graph showing a strong growth inhibitory effect against Staphylococcus aureus which is a food-mediated pathogen, the antimicrobial composition containing wheat germ extract of the present invention as an example of food Staphylococcus aureus in soy milk It is a graph showing the antimicrobial effect against.
  • Figure 2 is a graph showing a strong growth inhibitory effect against staphylococcus aureus, the main pathogen through which the antimicrobial composition containing wheat germ extract of the present invention is a food-borne pathogen to Staphylococcus aureus in milk as an example of food. It is a graph showing the antimicrobial effect against.
  • Figure 3 is a graph showing the results of the Staphylococcus aureus killing rate for the effect of killing the Staphylococcus aureus antibacterial composition containing the bran extract extracted by the method of extracting bran of one embodiment of the present invention.
  • Figure 4 is a graph showing a strong growth inhibitory effect against staphylococcus aureus, the main pathogen through which the antimicrobial composition containing wheat bran extract of the present invention as a food-borne pathogen to Staphylococcus aureus in soy milk as an example of food. It is a graph showing the antimicrobial effect against.
  • FIG. 5 is a graph showing a strong growth inhibitory effect against Staphylococcus aureus, which is a food-mediated pathogen, is an antimicrobial composition containing wheat bran extract of the present invention for Staphylococcus aureus in milk, which is an example of food. It is a graph showing the antimicrobial effect.
  • Wheat germ or wheat bran was purchased from Dong-A Won Co., Ltd. (Seoul, Korea) and used, and pathogens that are the antimicrobial and antiseptic test of wheat germ extract or wheat bran extract of the present invention (in the present invention, human or animal)
  • pathogens that are the antimicrobial and antiseptic test of wheat germ extract or wheat bran extract of the present invention (in the present invention, human or animal)
  • microorganisms such as viruses, bacteria, and fungi that cause disease
  • Salmonella typhimurium , Escherichia coli , Listeria monocytogenes , Staphylococcus aureus , and Bacillus Bacillus cereus , Pseudonomas aeruginosa , Candida albicans and Aspergillus niger
  • KCTC Korean Bank of Biotechnology for Type Cultures
  • nutritional medium for assaying antimicrobial activity (containing 0.03% beef extract and 0.05% peptone) was obtained from the BBL Microbiology System (Cockeysville, MD, USA.), And beef extract and peptone were obtained from Difco Laboratories (Detroit, MI, USA). Purchased from Other materials were purchased from Sigma Co., U.S.A., USA, unless otherwise noted.
  • the antimicrobial activity was detected by modified disk diffusion method (Bauer, AW, WM Kirby, JC Sherris, and M. Turck. 1966. Antibiotic susceptibility testing by a standardized single disk method.Am J. Clin. Pathol. 45: 493-496), to facilitate the diffusion of antimicrobial material and to analyze the antimicrobial activity of wheat germ extract or wheat bran extract of the present invention using agar well diffusion assay well known in the art.
  • agar well plates were prepared by cutting agar wells from agar medium sprayed with pathogens using a 7 mm diameter hollow tube, and applying a slight negative pressure to remove agar plugs.
  • the sensitivity of each pathogen to the test substance was determined by the formation of an inhibition zone after incubating the agar well plate for 18 hours at 37 ° C.
  • the experiments were conducted in triplicates and the results were expressed as mean values for the three measurements.
  • the minimum inhibitory concentration is Christoph et al. Christoph, M.S., P. Klaus, W. Annette, S. Erwin, and J. C. Simon. 1999. Antibacterial activity of hyperforin from St. John's wort, against multiresistant Staphylococcus aureus and gram-positive bacteria. The Lancet. 353: 2129-2131) and macrodilution test. Serial 2-fold dilutions of the test substance were prepared in dimethylsulfoxide, and 0.5 ml of dilution of test substance was added to 9.0 ml of nutrient medium. Salmonella cultured overnight in the diluted solution of each test substance thus obtained ( Salmonella typhimurium ), E.
  • Bacillus cereus bacteria Bacillus cereus 0.5 ml of the culture medium was injected. After 48 hours of incubation at 37 ° C. (but Bacillus cereus bacterium at 30 ° C.), a minimum inhibitory concentration was determined, which is the minimum concentration of test substance that results in no visible growth of pathogens (see Table 1 below).
  • the wheat bran extract of the present invention has a minimum inhibitory concentration of ⁇ 1.25 mg / ml, which is the minimum concentration of a test substance which does not show visible growth of Staphylococcus aureus , and the wheat bran extract is higher than wheat germ extract. It is confirmed that about 5 times lower, the antibacterial activity is about 5 times better.
  • the antimicrobial activity of wheat germ extract or wheat bran extract in Tables 2 to 9 of the following examples was measured by growth inhibition zone test compared to the positive control and the negative control, and the results are expressed in mm. .
  • the results shown in Tables 2 to 9 represent the mean value ⁇ standard deviation for the measured values obtained from a total of three experiments, and all the data in Tables 2 to 9 were statistical analysis programs [Statistical Package for Social Science (SPSS; SPSS Inc., Chicago, IL, USA).
  • Example 3 Antimicrobial efficacy test by wheat germ or wheat bran extract preparation and disk diffusion method
  • Example 3-1 Preparation of wheat germ or wheat bran extract and antibacterial efficacy test using chloroform
  • the prepared wheat germ was pulverized and 2.5 L of secondary distilled water was added to 100 g of the ground wheat germ, stirred for 1 hour, and then 1 L of chloroform (CHCl 3 ) solution was added to the solution for 30 minutes.
  • the solvent layer was recovered by stirring and centrifugation. At this time, the chloroform layer was removed after centrifugation, and the remaining filtrate layer was subjected to chloroform extraction three times in the same manner.
  • the chloroform layers were combined, washed with distilled water and concentrated at a maximum temperature of 40 ° C. using a vacuum evaporator. The concentrated material (6.2 g) was finally redissolved in chloroform and filtered through a 0.45- ⁇ m PTFE filter.
  • Salmonella (Salmonella typhimurium), as described in Example 2 by using the wheat germ extract obtained is filtered, treated according to the process as a sample, E. coli (Escherichia coli), Listeria monocytogenes Zenith (Listeria monocytogenes), Staphylococcus aureus (Staphylococcus aureus ) or 150 ⁇ l of the sample was put into each agar well plate in which Bacillus cereus was cultured, and the antimicrobial activity was measured using the well diffusion method (see Table 2).
  • E. coli Esscherichia coli
  • Listeria monocytogenes Zenith Listeria monocytogenes
  • Staphylococcus aureus Staphylococcus aureus
  • 150 ⁇ l of the sample was put into each agar well plate in which Bacillus cereus was cultured, and the antimicrobial activity was measured using the well diffusion method (see Table 2).
  • the prepared wheat bran was pulverized and 2.5 L of secondary distilled water was added to 100 g of the ground wheat bran, followed by stirring for 1 hour. Then, 1 L of chloroform (CHCl 3 ) solution was added again to the solution and re-stirred for 30 minutes. The solvent layer was recovered by centrifugation. At this time, the chloroform layer was removed after centrifugation, and the remaining filtrate layer was subjected to chloroform extraction three times in the same manner. The chloroform layers were combined, washed with distilled water and concentrated at a maximum temperature of 40 ° C. using a vacuum evaporator.
  • the concentrated material (5.8 g) was finally redissolved in chloroform and filtered through a 0.45- ⁇ m PTFE filter.
  • Salmonella typhimurium , Escherichia coli , Listeria monocytogenes , Staphylococcus aureus Or 150 ⁇ l of the sample was put into each agar well plate in which Bacillus cereus was cultured, and the antimicrobial activity was measured using the well diffusion method (see Table 3).
  • the wheat germ extract (1.5g fresh weight (FW; 150 ⁇ l)) of this Example 3-1 was analyzed by the antimicrobial activity of five food and cosmetic harmful bacteria, It can be seen that it exhibits even antimicrobial activity against Gram-positive and Gram-negative bacteria, especially Staphylococcus aureus (approximately 27.22 mm growth inhibition zone) and Bacillus cereus (approximately 12.99 mm growth). It can be seen that it has a strong growth inhibitory effect on the zone of inhibition).
  • the wheat bran extract (1.5g fresh weight (FW; / 150 ⁇ l) of this Example 3-1) of the antimicrobial activity analysis results for five harmful bacteria and food products , It can be seen that it shows even antimicrobial activity against Gram-positive and Gram-negative bacteria, in particular Staphylococcus aureus (approximately 15.91mm growth inhibition zone) and Bacillus cereus (approximately 12.94mm It can be seen that it has a strong growth inhibitory effect on the growth inhibition zone).
  • wheat germ extract or wheat bran extract of the present invention can be used as a useful antimicrobial material for food poisoning prevention. That is, the wheat germ extract or wheat bran extract of the present invention can be confirmed that the material is applicable as an antiseptic for the safe production of food or drink and can be used as an antibacterial and antibiotic.
  • Example 3-2 Preparation of wheat germ or wheat bran extract and antibacterial efficacy test using methylene chloride
  • the prepared wheat germ was pulverized, and 2.5 L of secondary distilled water was added to 100 g of the ground wheat germ, followed by stirring for 1 hour. Then, 1 L of methylene chloride (CH 2 Cl 2 ) solution was further added to the solution. The solvent layer was recovered by restirring for minutes and centrifuging. At this time, the methylene chloride layer was removed after centrifugation, and the remaining filtrate layer was subjected to methylene chloride extraction three times in the same manner. The methylene chloride layers were combined, washed with distilled water and concentrated at a maximum temperature of 40 ° C. using a vacuum evaporator.
  • CH 2 Cl 2 methylene chloride
  • the concentrated material (5.9 g) was finally redissolved in methylene chloride and filtered through a 0.45- ⁇ m PTFE filter.
  • Salmonella Salmonella (Salmonella typhimurium), as described in Example 2 by using the wheat germ extract obtained is filtered, treated according to the process as a sample, E. coli (Escherichia coli), Listeria monocytogenes Zenith (Listeria monocytogenes), Staphylococcus aureus (Staphylococcus aureus ) or 150 ⁇ l of the sample was put into each agar well plate in which Bacillus cereus was cultured, and the antimicrobial activity was measured using the well diffusion method (see Table 2 above).
  • the prepared wheat bran was pulverized and stirred for 1 hour by adding 2.5 L of secondary distilled water to 100 g of the crushed wheat bran, and then 1 L of methylene chloride (CH 2 Cl 2 ) solution was added to the solution for 30 minutes.
  • the solvent layer was recovered by restirring and centrifugation. At this time, the methylene chloride layer was removed after centrifugation, and the remaining filtrate layer was subjected to methylene chloride extraction three times in the same manner.
  • the methylene chloride layers were combined, washed with distilled water and concentrated at a maximum temperature of 40 ° C. using a vacuum evaporator.
  • the concentrated material (5.4 g) was finally redissolved in methylene chloride and filtered through a 0.45- ⁇ m PTFE filter.
  • Salmonella typhimurium Escherichia coli , Listeria monocytogenes , Staphylococcus aureus
  • 150 ⁇ l of each sample was put into an agar well plate in which Bacillus cereus was cultured, and the antimicrobial activity was measured using the well diffusion method (see Table 3 above).
  • the wheat germ extract (1.5g fresh weight (FW; 150 ⁇ L)) of this Example 3-2 was analyzed by the antimicrobial activity of five harmful food and cosmetic bacteria, It can be seen that it exhibits even antimicrobial activity against Gram-positive and Gram-negative bacteria, in particular Staphylococcus aureus (approximately 29.59 mm growth inhibition zone) and Bacillus cereus (approximately 14.49 mm It can be seen that it has a strong growth inhibitory effect on the growth inhibition zone).
  • wheat germ extract or wheat bran extract of the present invention can be used as a useful antimicrobial material for food poisoning prevention. That is, the wheat germ extract or wheat bran extract of the present invention can be confirmed that the material is applicable as an antiseptic for the safe production of food or drink and can be used as an antibacterial and antibiotic.
  • the prepared wheat germ was pulverized, and 2.5 L of secondary distilled water was added to 100 g of the ground wheat germ, followed by stirring for 1 hour, and then 1 L of ethyl acetate (CH 3 COOC 2 H 5 ) solution was added to the solution. The mixture was restirred for 30 minutes and the solvent layer was recovered by centrifugation. At this time, the ethyl acetate layer was removed after centrifugation, and the remaining filtrate layer was extracted three times with the same method. The ethyl acetate layers were combined, washed with distilled water and concentrated at a maximum temperature of 40 ° C. using a vacuum evaporator.
  • the concentrated material (5.5 g) was finally redissolved in ethyl acetate and filtered through a 0.45- ⁇ m PTFE filter.
  • Salmonella Salmonella (Salmonella typhimurium), as described in Example 2 by using the wheat germ extract obtained is filtered, treated according to the process as a sample, E. coli (Escherichia coli), Listeria monocytogenes Zenith (Listeria monocytogenes), Staphylococcus aureus (Staphylococcus aureus ) or 150 ⁇ l of samples were put into each agar well plate in which Ba cillus cereus was cultured, and the antimicrobial activity was measured using the well diffusion method (see Table 4).
  • the prepared wheat bran was pulverized and 2.5 L of secondary distilled water was added to 100 g of the crushed wheat bran, followed by stirring for 1 hour. Then, 1 L of ethyl acetate (CH 3 COOC 2 H 5 ) was added to the solution. The solvent layer was recovered by restirring for minutes and centrifuging. At this time, the ethyl acetate layer was removed after centrifugation, and the remaining filtrate layer was extracted three times with the same method. The ethyl acetate layers were collected, washed with distilled water and concentrated at a maximum temperature of 40 ° C. using a vacuum evaporator.
  • the concentrated material (4.8 g) was finally redissolved in ethyl acetate and filtered through a 0.45- ⁇ m PTFE filter.
  • Salmonella typhimurium , Escherichia coli , Listeria monocytogenes , Staphylococcus aureus Or 150 ⁇ l of the sample was put into each agar well plate in which Bacillus cereus was cultured, and the antimicrobial activity was measured using the well diffusion method (see Table 5).
  • the wheat bran extract (1.5g fresh weight (FW; / 150 ⁇ l) of this Example 3-3) is the antimicrobial activity analysis results for five harmful bacteria and food products , It can be seen that it shows even antimicrobial activity against Gram-positive and Gram-negative bacteria, especially Staphylococcus aureus (approximately 19.42mm growth inhibition zone) and Bacillus cereus (approximately 13.89mm It can be seen that it has a strong growth inhibitory effect on the growth inhibition zone).
  • wheat germ extract or wheat bran extract of the present invention can be used as a useful antimicrobial material for food poisoning prevention. That is, the wheat germ extract or wheat bran extract of the present invention can be confirmed that the material is applicable as an antiseptic for the safe production of food or drink and can be used as an antibacterial and antibiotic.
  • the prepared wheat germ was pulverized and 1 L of diethylene glycol monoethyl ether (carbitol) solution was added to 100 g of the ground wheat germ, stirred for 30 minutes, and the solvent layer was recovered by centrifugation. At this time, the diethylene glycol monoethyl ether (carbitol) layer was removed after centrifugation, and the remaining precipitate was subjected to diethylene glycol monoethyl ether (carbitol) extraction three times in the same manner. The diethylene glycol monoethylether (carbitol) layers were collected and concentrated at a maximum temperature of 40 ° C. The concentrated material (4.9 g) was finally filtered through a 0.45- ⁇ m PTFE filter.
  • diethylene glycol monoethyl ether (carbitol) solution was added to 100 g of the ground wheat germ, stirred for 30 minutes, and the solvent layer was recovered by centrifugation. At this time, the diethylene glycol monoethyl ether (carbit
  • Salmonella (Salmonella typhimurium), as described in Example 2 by using the wheat germ extract obtained is filtered, treated according to the process as a sample, E. coli (Escherichia coli), Listeria monocytogenes Zenith (Listeria monocytogenes), Staphylococcus aureus (Staphylococcus aureus ) or 150 ⁇ l of samples were put into each agar well plate in which Bacillus cereus was cultured, and the antimicrobial activity was measured using the well diffusion method (see Table 6).
  • E. coli Esscherichia coli
  • Listeria monocytogenes Zenith Listeria monocytogenes
  • Staphylococcus aureus Staphylococcus aureus
  • 150 ⁇ l of samples were put into each agar well plate in which Bacillus cereus was cultured, and the antimicrobial activity was measured using the well diffusion method (see Table 6).
  • the prepared wheat bran was pulverized, and 1 L of diethylene glycol monoethyl ether (carbitol) solution was added to 100 g of the crushed wheat bran, stirred for 30 minutes, and the solvent layer was recovered by centrifugation. At this time, the diethylene glycol monoethyl ether (carbitol) layer was removed after centrifugation, and the remaining precipitate was subjected to diethylene glycol monoethyl ether (carbitol) extraction three times in the same manner. Diethylene glycol monoethylether (carbitol) layers were collected and concentrated at a maximum temperature of 40 ° C. The concentrated material (7.2 g) was finally filtered through a 0.45- ⁇ m PTFE filter.
  • diethylene glycol monoethyl ether (carbitol) solution was added to 100 g of the crushed wheat bran, stirred for 30 minutes, and the solvent layer was recovered by centrifugation. At this time, the diethylene glycol monoethyl ether
  • bran extract obtained by filtration according to the above process as a sample, as described in Example 2 Salmonella typhimurium , Escherichia coli , Listeria monocytogenes , Staphylococcus aureus ), Or 150 ⁇ l of each sample was put into an agar well plate in which Bacillus cereus was cultured, and the antimicrobial activity was measured using the well diffusion method (see Table 7).
  • the wheat germ extract (1.5g fresh weight (FW; 150 ⁇ l)) of this Example 3-4 was analyzed by the antimicrobial activity of five harmful food and cosmetic bacteria, It can be seen that it exhibits even antimicrobial activity against Gram-positive and Gram-negative bacteria, especially Staphylococcus aureus (approximately 25.59 mm growth inhibition zone) and Bacillus cereus (approximately 13.49 mm growth). It can be seen that it has a strong growth inhibitory effect on the zone of inhibition).
  • the wheat bran extract 1.5g fresh weight (FW; 150 ⁇ l) of this Example 3-4) of the antimicrobial activity analysis results for five food and cosmetic harmful bacteria , It can be seen that it shows even antimicrobial activity against Gram-positive and Gram-negative bacteria, in particular Staphylococcus aureus (approximately 18.18mm growth inhibition zone) and Bacillus cereus (approximately 15.19mm It can be seen that it has a strong growth inhibitory effect on the growth inhibition zone).
  • wheat germ extract or wheat bran extract of the present invention can be used as a useful antimicrobial material for food poisoning prevention. That is, the wheat germ extract or wheat bran extract of the present invention can be confirmed that the material is applicable as an antiseptic for the safe production of food or drink and can be used as an antibacterial and antibiotic.
  • Example 3-5 Preparation of Wheat Germ or Wheat Bran Extract and Antimicrobial Efficacy Test Using Supercritical Fluid Extraction Method
  • the supercritical fluid extraction method used in this example is Extraction method using a supercritical fluid that is a fluid that exceeds the critical temperature and boundary pressure.
  • the supercritical fluid extraction method is more efficient than the liquid extraction method. Since the solvent power is strongly dependent on the density, the extraction conditions suitable for the target material can be obtained by changing the operating temperature or the pressure. Generally, inert, tasteless, odorless, and harmless carbon dioxide is used at room temperature, so it can be safely used in foods and medicines. At normal pressure, only the extract component is easily obtained because the fluid vaporizes.
  • the antimicrobial activity of the wheat germ extract or wheat bran extract extracted using the supercritical fluid extraction method was evaluated by the disk diffusion method (see Table 8 and Table 9).
  • Example 2 the wheat germ extract or wheat bran extract obtained by filtration according to the above procedure was used as a sample, as described in Example 2, Salmonella typhimurium , Escherichia coli , Listeria monocytogenes , 150 ⁇ l of each sample was added to agar well plates in which Staphylococcus aureus or Bacillus cereus were cultured, and the antimicrobial activity was measured using the well diffusion method (see Table 8 and Table 9).
  • the wheat germ extract (1.5g fresh weight (FW; 150 ⁇ l)) of the present Example 3-5 was analyzed by the antimicrobial activity of five harmful bacteria and food, cosmetics, It can be seen that it exhibits even antimicrobial activity against Gram-positive and Gram-negative bacteria, especially Staphylococcus aureus (approximately 26.77 mm growth inhibition zone) and Bacillus cereus (approximately 13.01 mm growth). It can be seen that it has a strong growth inhibitory effect on the zone of inhibition).
  • the wheat bran extract (1.5g fresh weight (FW; / 150 ⁇ l) of this Example 3-5) is the antimicrobial activity analysis results for five food and cosmetic harmful bacteria , It can be seen that it shows even antimicrobial activity against Gram-positive and Gram-negative bacteria, especially Staphylococcus aureus (approximately 29.84mm growth inhibition zone) and Bacillus cereus (approximately 13.43mm It can be seen that it has a strong growth inhibitory effect on the growth inhibition zone).
  • wheat germ extract or wheat bran extract of the present invention can be used as a useful antimicrobial material for food poisoning prevention. That is, the wheat germ extract or wheat bran extract of the present invention can be confirmed that the material is applicable as an antiseptic for the safe production of food or drink and can be used as an antibacterial and antibiotic.
  • the wheat germ extract of the present invention has a strong growth inhibitory effect against Staphylococcus aureus concentration-dependent
  • the results shown in Figures 4 and 5 it was confirmed that wheat bran extract of the present invention also has a strong growth inhibitory effect against Staphylococcus aureus concentration-dependent. From this, it could be confirmed once again that the wheat germ extract or wheat bran extract of the present invention is applicable as an antiseptic for the safe production of foods or beverages and can be used as an antibacterial and antibiotic.
  • Staphylococcus aureus ( Staphylococcus aureus ) was added to the liquid medium medium of the concentration of about 10 4 CFU / ml by the concentration of the wheat bran extract of the present invention was incubated for 24 hours to measure the killing of bacteria. As a result, as shown in Figure 3, it was confirmed that the killing of Staphylococcus aureus occurred within one hour after adding the bran extract of the present invention to the Staphylococcus aureus liquid medium.
  • Staphylococcus aureus Escherichia coli , Pseudonomas aeruginosa , Candida albicans and Aspergillus niger were incubated at 37 ° C and 25 ° C for 18 hours and then 10 -3 After diluting and adding 1 ml of each culture to 9 g of the cosmetic base (Table 12) sample, the mixture was stirred, and then the wheat germ extract or wheat bran extract of the present invention was added to each sample at a concentration of 0.5-2.0% to 25 ° C. The number of bacteria in each sample was measured on days 0, 7, 14 for the antiseptic test of the stored samples.
  • the wheat germ extract or wheat bran extract of the present invention can be used as food and cosmetics as a safe and inexpensive natural antimicrobial agent against food-borne pathogens.
  • wheat germ and wheat bran are inexpensive and easily available in large quantities as a by-product of the flour production process, so that wheat germ extract or wheat bran extract can be widely used in food, beverages, cosmetics, etc.
  • the present invention uses chloroform, methylene chloride, ethyl acetate or diethylene glycol monoethyl ether as an example for extracting antimicrobial substances from wheat germ or wheat bran, but the present invention is not limited to these solvents, but various food fields. And of course, it does not exclude known organic solvents that can be used for the extraction of active ingredients in the medical field.

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Abstract

La présente invention concerne une composition antimicrobienne contenant un extrait de germe de blé ou un extrait de son de blé ayant des effets antimicrobiens, qui peut remplacer des agents de conservation ou des antibiotiques ajoutés aux aliments, aux produits cosmétiques, aux aliments pour animaux, ou analogues. Le germe de blé ou son de blé est un sous-produit de la production en série de farine, et peut donc être obtenu facilement et à un faible coût. La composition antimicrobienne selon la présente invention utilisant du germe de blé ou du son de blé peut être grandement utilisée dans des aliments, des boissons, des produits cosmétiques, ou analogues, et peut être également utilisée comme substance antimicrobienne pouvant remplacer des antibiotiques dans des aliments pour animaux, qui est devenu un sujet de grande importance. Par conséquent, la composition antimicrobienne selon la présente invention peut prévenir et traiter des maladies pathogènes provoquées par des aliments ou analogues et peut être utilisée comme une nouvelle substance antimicrobienne naturelle pour la prévention d'un accroissement de tolérance aux antibiotiques. La composition antimicrobienne selon la présente invention peut être utilisée pour une large gamme de domaines comprenant ceux d'aliments pour animaux, de produits alimentaires, de boissons, de produits cosmétiques, d'emballage de produits alimentaires entre autres.
PCT/KR2011/005541 2010-07-30 2011-07-27 Nouvelle composition antimicrobienne, et son procédé de préparation WO2012015242A2 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015071413A1 (fr) * 2013-11-15 2015-05-21 Universitat Autonoma De Barcelona Extrait soluble de son de blé utilisé comme agent anti-biofilm
CN111493101A (zh) * 2020-05-05 2020-08-07 广州市美驰化妆品有限公司 采用植物油或合成油脂浸泡法从药用植物中制备消毒剂及应用

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
HARRY HUMFELD: 'Antibiotic Activity of the Wheat Bran Fatty-Acid-like Constituents of Wheat Bran.' J.BACTERIOL. vol. 54, no. 4, 1947, pages 513 - 517 *
STIPKOVITS, L. ET AL.: 'Testing the Efficacy of Fermented Wheat Germ Extract Against Mycoplasma gallisepticum Infection of Chickens' POULT. SCI. vol. 83, no. 11, 2004, pages 1844 - 1848 *
ZAHI AI-HUA ET AL.: 'Study on Antibacteriality of The Flavonids of Wheat Germ.' JOURNAL OF HEILONGJIANG AUGUST FIRST LAND RECLAMATION UNIVERSITY 2005, *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015071413A1 (fr) * 2013-11-15 2015-05-21 Universitat Autonoma De Barcelona Extrait soluble de son de blé utilisé comme agent anti-biofilm
CN111493101A (zh) * 2020-05-05 2020-08-07 广州市美驰化妆品有限公司 采用植物油或合成油脂浸泡法从药用植物中制备消毒剂及应用

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