KR101807387B1 - A method for the bacterial inactivation by treating fruit extracts and essential oil, and a composition therefor - Google Patents
A method for the bacterial inactivation by treating fruit extracts and essential oil, and a composition therefor Download PDFInfo
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Classifications
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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
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/36—Rutaceae [Rue family], e.g. lime, orange, lemon, corktree or pricklyash
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
본 발명은 깔라만시, 레몬 및 라임에서 선택되는 하나 이상의 과일로부터의 과일즙; 및 카바크롤 및 티몰에서 선택되는 하나 이상의 에센셜오일을 혼합처리하여 미생물을 사멸시키는 방법 및 이를 위한 조성물에 관한 것이다.The present invention relates to fruit juice from one or more fruits selected from calamansi, lemons and lime; And at least one essential oil selected from carbacol and thymol are mixed with each other to kill the microorganism, and a composition therefor.
Description
본 발명은 미생물을 사멸시키는 방법 및 이를 위한 조성물에 관한 것이다. The present invention relates to a method for killing microorganisms and a composition therefor.
시트러스(citrus) 과일은 높은 비타민 C 함량을 갖는 산성 과일로 특유의 상큼한 맛뿐만 아니라 항산화 작용, 독소제거 등의 건강상의 이점 때문에 전 세계적으로 꾸준한 인기를 끌며 가장 많이 소비되는 과일류 중 하나이다. 이는 음료(주스, 에이드 등)의 형태로도 많이 섭취되고 즙의 형태로 여러 식품, 양념 등에 첨가되어 향과 맛을 더해주는 재료로 사용된다. 시트러스 과일은 유기산인 시트르산(citric acid)을 함유하여 높은 산성을 갖기 때문에 일반적으로 낮은 pH를 갖는 과일주스는 위해 세균으로부터 안전하고, 과일즙은 잠재적 위해 식품(Potentially Hazardous Foods)에 첨가되어 미생물학적 품질을 향상시킬 수 있다고 알려져 있다. 하지만 산 적응(acid adaptation) 균주의 등장에 따라 과일주스에서 식중독 세균이 검출되고(표 1), 식품에서 과일즙 첨가로 효과적으로 식중독 세균을 저감하지 못하는 경우가 발생하였다(표 2). 따라서 과일주스와 과일즙이 첨가된 잠재적 위해 식품의 미생물학적 품질과 안전성을 향상시키기 위하여 시트러스 과일즙의 항균력을 향상시킬 수 있는 기술이 요구된다.Citrus fruit is an acidic fruit with a high vitamin C content and is one of the most popular and most consumed fruit species in the world because of its unique refreshing taste as well as its health benefits such as antioxidant activity and elimination of toxins. It is also consumed in the form of beverage (juice, aide, etc.) and is used as a material to add flavor and taste to various foods, spices and the like in the form of juice. Because citrus fruit contains citric acid, which is an organic acid, it has high acidity, fruit juices with low pH are generally safe from bacteria and fruit juices are added to potentially hazardous foods, Can be improved. However, due to the appearance of acid adaptation strains, foodborne pathogens were detected in fruit juices (Table 1) and food poisoning bacteria could not be effectively reduced by adding fruit juice (Table 2). Therefore, there is a need for a technique that can enhance the antimicrobial activity of citrus fruit juice in order to improve the microbiological quality and safety of potentially harmful food added with fruit juice and fruit juice.
최근 천연 식품 첨가제에 대한 소비자의 요구가 증가함에 따라 식물과 같은 천연자원에서 유래한 식품 보존제가 인기를 얻고 있다. 식물에서 추출한 방향족 물질인 에센셜 오일(essential oil)은 이러한 소비자의 요구에 맞춰 식품 첨가제 및 보존제 등으로 식품 시장에서 다양하게 사용되고 있다. 에센셜 오일은 다수의 선행연구에서 다양한 식중독 세균(바실러스 속 균, 대장균, 장출혈성 대장균 O157:H7, 리스테리아 속 균, 살모넬라 속 균, 황색포도상구균, 시겔라 속 균 등)에 대해서 항균작용을 하는 것으로 알려졌고 미국과 유럽에서 식품 첨가제로 허가되었다. 하지만 에센셜 오일이 식품 내에서 항균력을 나타내기 위해서는 고농도가 요구되며 천연 물질이므로 합성 첨가제보다 경제적으로 부담이 있기 때문에 다른 식품 보존료나 식품 보존 기술과 에센셜 오일을 조합하여 세균을 제어하기 위한 연구가 요구된다.Recently, as consumer demands for natural food additives have increased, food preservatives derived from natural sources such as plants have become popular. Essential oil, an aromatic material extracted from plants, is widely used in the food market as a food additive and a preservative to meet the needs of such consumers. Essential oils are antimicrobial to many foodborne pathogens (Bacillus, Escherichia coli, Enterohemorrhagic Escherichia coli O157: H7, Listeria, Salmonella, Staphylococcus aureus, and Shigella) in a number of previous studies And was approved as a food additive in the United States and Europe. However, since essential oils are required to have a high concentration in order to exhibit antimicrobial activity in foods, it is more economical than synthetic additives because it is a natural substance, so research for controlling bacteria by combining other food preservatives, food preservation techniques and essential oils is required .
따라서 본 발명자는 천연항균물질인 에센셜 오일과 시트러스 과일즙을 이용하여 일반 식중독 세균뿐만 아니라 산적응 식중독 세균까지 효과적으로 제어할 수 있는 신기술을 개발하고자 연구하여, 본 발명을 완성하였다.Therefore, the present inventors have completed the present invention by studying to develop a new technology capable of effectively controlling not only general food poisoning bacteria but also acid-sensitive food poisoning bacteria by using natural antimicrobial substances, essential oil and citrus fruit juice.
상술한 문제점을 해결하기 위하여, 본 발명은 천연항균물질인 에센셜 오일과 시트러스 과일즙을 이용하여 일반 식중독 세균뿐만 아니라 산적응 식중독 세균까지 효과적으로 제어할 수 있는 방법 및 조성물을 제공하는 것을 목적으로 한다.In order to solve the above problems, it is an object of the present invention to provide a method and composition for effectively controlling not only general food poisoning bacteria but also acid-adapted food poisoning bacteria using essential oil and citrus fruit juice which are natural antibacterial substances.
본 발명은, 본 발명은 깔라만시, 레몬 및 라임에서 선택되는 하나 이상의 과일로부터의 과일즙; 및 카바크롤 및 티몰에서 선택되는 하나 이상의 에센셜오일을 혼합처리하여 미생물을 사멸시키는 방법 및 이를 위한 조성물을 제공한다.The present invention relates to a fruit juice from one or more fruits selected from calamansi, lemons and lime; And at least one essential oil selected from carbacol and thymol are mixed with each other to kill the microorganisms, and a composition therefor.
본 발명의 미생물을 사멸시키는 방법 및 이를 위한 조성물을 사용하면, 시트러스 과일즙과 식물에서 추출한 천연항균물질인 에센셜 오일의 항균 시너지 효과를 통해 단시간 안에 효과적으로 미생물을 제어할 수 있을 뿐 아니라, 식품에 매우 안전하게 적용가능 하다.The method of killing the microorganisms of the present invention and the composition for the microorganism of the present invention can effectively control the microorganisms in a short time by virtue of the antibacterial synergy of citrus fruit juice and essential oil which is a natural antibacterial substance extracted from plants, It is safe to apply.
도 1은 일반 배양 E. coli O157:H7 에 대한 카바크롤 함유 과일즙 용액의 미생물 사멸 효과를 나타낸다.
도 2는 일반 배양 E. coli O157:H7 에 대한 티몰 함유 과일즙 용액 의 미생물 사멸 효과를 나타낸다.
도 3은 산적응 배양 E. coli O157:H7 에 대한 카바크롤 함유 과일즙 용액 의 미생물 사멸 효과를 나타낸다.
도 4는 산적응 배양 E. coli O157:H7 에 대한 티몰 함유 과일즙 용액 의 미생물 사멸 효과를 나타낸다.
도 5는 일반 배양 S. Typhimurium 에 대한 카바크롤 함유 과일즙 용액 의 미생물 사멸 효과를 나타낸다.
도 6은 일반 배양 S. Typhimurium에 대한 티몰 함유 과일즙 용액 의 미생물 사멸 효과를 나타낸다.
도 7은 산적응 배양 S. Typhimurium에 대한 카바크롤 함유 과일즙 용액 의 미생물 사멸 효과를 나타낸다.
도 8은 산적응 배양 S. Typhimurium에 대한 티몰 함유 과일즙 용액 의 미생물 사멸 효과를 나타낸다.
도 9는 일반 배양 L. monocytogenes 에 대한 카바크롤 함유 과일즙 용액 의 미생물 사멸 효과를 나타낸다.
도 10은 일반 배양 L. monocytogenes 에 대한 티몰 함유 과일즙 용액 의 미생물 사멸 효과를 나타낸다.
도 11은 산적응 배양 L. monocytogenes 에 대한 카바크롤 함유 과일즙 용액 의 미생물 사멸 효과를 나타낸다.
도 12는 산적응 배양 L. monocytogenes 에 대한 티몰 함유 과일즙 용액의 미생물 사멸 효과를 나타낸다.
도 13은 산적응 배양 E. coli O157:H7 에 대한 카바크롤 함유 유기산, 과일즙 용액 의 미생물 사멸 효과를 나타낸다.
도 14: 산적응 배양 E. coli O157:H7 에 대한 티몰 함유 유기산, 과일즙 용액 의 미생물 사멸 효과를 나타낸다.Figure 1 shows the microbial killing effect of carbacol-containing fruit juice solution on normal cultured E. coli O157: H7.
Figure 2 shows the microbial killing effect of the thymol-containing fruit juice solution on the normal cultured E. coli O157: H7.
Figure 3 shows the microbial killing effect of carbacol-containing fruit juice solutions on acid adapted cultures E. coli O157: H7.
Figure 4 shows the microbial killing effect of the thymol-containing fruit juice solution on acid adapted cultures E. coli O157: H7.
5 is a general culture S. This shows the microbial killing effect of carbacol-containing fruit juice solution against Typhimurium.
6 is a general culture S. The microbial killing effect of the thymol-containing fruit juice solution on Typhimurium is shown.
FIG. 7 shows the results of the acid-adapted culture S. This shows the microbial killing effect of carbacol-containing fruit juice solution against Typhimurium.
FIG. 8 shows the results of the acid-adapted culture S. The microbial killing effect of the thymol-containing fruit juice solution on Typhimurium is shown.
Figure 9 shows the microbial killing effect of carbacol-containing fruit juice solution on the general culture L. monocytogenes .
10 shows the microbial killing effect of the thymol-containing fruit juice solution on the general culture L. monocytogenes .
Figure 11 shows the microbial killing effect of cupricol-containing fruit juice solution on acid-adapted cultured L. monocytogenes .
Figure 12 shows the microbial killing effect of the thymol-containing fruit juice solution on acid-adapted cultured L. monocytogenes .
Figure 13 shows the microbial killing effect of carbacol-containing organic acid, fruit juice solution on acid adapted cultures E. coli O157: H7.
Fig. 14: Microbial killing effect of thiom-containing organic acid, fruit juice solution on acid-adapted cultured E. coli O157: H7.
이하, 본 발명을 보다 상세하게 설명한다.Hereinafter, the present invention will be described in more detail.
본 발명은, 과일즙과 에센셜오일을 포함하는 미생물 사멸을 위한 조성물 및 과일즙과 에센셜오일을 혼합처리하여 미생물을 사멸시키는 방법에 대한 것으로, 과일즙과 에센셜오일을 함께 처리하는 경우, 미생물 사멸효과에 있어 각각의 단독처리의 효과나 이들 효과의 조합에 의해 예상되는 효과에 비해 미생물 사멸효과가 현저히 상승됨을 실험적으로 확인하여 본 발명을 완성하였다. The present invention relates to a composition for killing microorganisms comprising fruit juice and essential oil, and a method for killing microorganisms by mixing fruit juice and essential oil. When fruit juice and essential oil are treated together, microorganism killing effect The inventors of the present invention have experimentally confirmed that the microbial killing effect is remarkably increased in comparison with the effects expected by the effects of each single treatment and the combination of these effects.
본 발명의 과일즙은, 시트러스 과일즙일 수 있다. 상기 시트러스 과일즙은 과일즙 중 유일하게 식품에 첨가되기 때문에 다양한 식품류에 적용하여 식품의 미생물학적 안전성을 높이는 데 이용될 수 있다. The fruit juice of the present invention may be a citrus fruit juice. Since the citrus fruit juice is the only fruit juice added to the food, it can be applied to various foods to enhance the microbiological safety of the food.
또한, 본 발명의 에센셜 오일은 미국 식품의약품안전청이 발표한 GRAS (Generally Recognized as Safe) 물질로 등재되어 있기 때문에 식품첨가물로 사용이 가능하므로 본 발명의 미생물 사멸 방법은 매우 안전한 방법으로 식품에 적용가능하다. Since the essential oil of the present invention is listed as a GRAS (Generally Recognized as Safe) substance announced by the US Food and Drug Administration, the microbial death method of the present invention can be applied to foods in a very safe manner Do.
본 발명의 상기 과일즙은, 깔라만시, 레몬 및 라임에서 선택되는 하나 이상의 과일로부터 제조된 것 일 수 있고, 상기 에센셜오일은 카바크롤 및 티몰에서 선택되는 하나 이상일 수 있다. The fruit juice of the present invention may be one prepared from one or more fruits selected from calamansi, lemon and lime, and the essential oil may be one or more selected from carbacrol and thymol.
상기 미생물은 식중독 미생물일 수 있으며, 크로모박테리움 바일레큠 (C. violaceum), 슈도모나스 애루지노사 (P. aeruginosa), 대장균 (E. coli), 파스튜렐라 헤몰리티카 (P. hemolytica), 황색 포도상구균 (S. aureus), 살모넬라 콜라라수이스 (S. cholerasuis). 리스테리아 모노사이토제네스 (L. monocytogenes), 살모넬라 티피뮤리움 (S. Typhimurium), 캠필로박터 제주니(Campylobacter jejuni), 여시니아 엔테로콜리티카(Yersinia enterocolitica), 크로노박터균(Cronobacter spp.), 이질균(Shigella spp.), 비브리오균(Vibrio spp.), 클로스트리디움 속균(Clostridium perfringens , Clostridium botulinum) 및 바실러스균(Bacillus spp.)에서 선택되는 하나 이상일 수 있다. 또한, 상기 대장균 (E. coli)은 장출혈성대장균(E. coli O157:H7) 일 수 있다. The microorganisms may be microbial food poisoning, chromotherapy tumefaciens Vejle kyum (C. violaceum), Pseudomonas her labor Rouge (P. aeruginosa), Escherichia coli (E. coli), par stew Relais H. Morley Utica (P. hemolytica) , Staphylococcus aureus ( S. aureus ), Salmonella cola raisuis ( S. cholerasuis ). L. monocytogenes jeneseu (L. monocytogenes), Salmonella typhimurium (S. Typhimurium), Campylobacter Jeju Needle (Campylobacter jejuni , Yersinia enterocolitica ), Cronobacter spp.), Shigella (Shigella spp.), Vibrio bacteria (Vibrio spp.), Clostridium perfringens (Clostridium botulinum ), and Bacillus spp. It can be: (H7 E. coli O157) In addition, the Escherichia coli (E. coli) Chapter hemorrhagic Escherichia coli.
상기 과일즙은 10 내지 20%의 농도일 수 있으며, 이는 과일주스에 포함되는 과일즙 농도와 식품에 첨가되는 수준의 농도를 고려할 때 가장 바람직한 범위이다. The fruit juice may be a concentration of 10 to 20%, which is the most preferable range considering the concentration of the fruit juice contained in the fruit juice and the level added to the food.
상기 과일즙 및 상기 에센셜오일을 혼합하여 제조되는 용액의 에센셜 오일의 최종 농도가 약 3 mM(약 0.047%)일 수 있으며, 바람직하게는 1 내지 2 mM(0.016 내지 0.031%)일 수 있다. The final concentration of the essential oil in the solution prepared by mixing the fruit juice and the essential oil may be about 3 mM (about 0.047%), preferably about 1 to 2 mM (0.016 to 0.031%).
본 발명의 미생물을 사멸하기 위한 방법은, 상기 설명한 사항 이외에 미생물 사멸을 위한 공지의 기술적 단계를 포함 가능하며, 본 발명의 미생물 사멸을 위한 조성물 역시 상기 설명한 사항 이외에 미생물 사멸을 위한 공지의 구성을 포함할 수 있다. The method for killing microorganisms according to the present invention may include known technical steps for killing microorganisms other than those described above. The composition for killing microorganisms of the present invention may also include a known composition for killing microorganisms can do.
이하, 본 발명을 더욱 상세하게 설명하기 위하여 실시예를 첨부한다. 하기 실시예는 본 발명의 이해를 보다 용이하게 하기 위하여 제공되는 것으로, 본 발명의 범위를 한정하는 것은 아니다. Hereinafter, embodiments will be described in order to explain the present invention in more detail. The following examples are provided to further facilitate understanding of the present invention and are not intended to limit the scope of the present invention.
참조예: 제어대상 식중독균Reference Example: Controlled food poisoning bacteria
시트러스 과일즙의 산도가 높다는 점에 착안하여 대표적인 내산성 식중독 세균인 Escherichia coli O157:H7 (ATCC 35150, 43889, 43895)과 주요 식중독 세균인 살모넬라 타이피뮤리움 (Salmonella Typhimurium ) (ATCC 19575, 43174, DT104), 리스테리아 모노사이토게네스 ( Listeria monocytogenes ) (ATCC 19115, 19117, 19119)를 실험대상 식중독 세균으로 선정하였으며, 개발 기술의 적용 가능성을 보다 넓게 제시하기 위하여 일반적인 환경에서 배양한 균(tryptic soy broth에 분주, 37℃, 24시간: 일반배양균)과 산적응(acid adaptation) 배양(일반배양균을 1% glucose를 첨가한 tryptic soy broth에 분주, 37℃, 24시간: 산적응배양균)에 의해 높은 내산성이 유도된 세포를 제조하여 실험에 사용하였다. 배양한 각 균주는 50 ml 플라스틱 튜브(Becton Diskinson, Franklin Lakes, NJ, USA)에 혼합한 후 3,000×g에서 15분간 원심분리 하였고(Centra-CL2, Needham Heights, MA, USA) 상등액을 따라낸 후 동량의 0.85% 멸균 생리식염수를 첨가하여 미생물 세포를 2회 세척함을 통해 균액(cell suspension)을 제조하였다.In view of the high acidity of citrus fruit juices, Escherichia , a representative acid-fast food poisoning bacterium coli O157: H7 (ATCC 35150, 43889, 43895) and the major food poisoning bacteria Salmonella Solarium typhimurium (Salmonella Typhimurium) (ATCC 19575, 43174, DT104), Listeria Monocytogenes to Ness (Listeria monocytogenes) (ATCC were selected for 19 115, 19 117, 19 119), the subjects of food poisoning bacteria, a culture in a typical environment to address wider than the applicability of the developed technology (frequency division in tryptic soy broth, 37 ℃, 24 hours: General Cultured cells) and acid adaptation cultures (normal cultures were dispensed in tryptic soy broth supplemented with 1% glucose at 37 ° C for 24 hours: acid-adapted cultures) to produce highly acid-labile cells Was used for the experiment. The cultures were mixed with 50 ml plastic tubes (Becton Diskinson, Franklin Lakes, NJ, USA) and centrifuged at 3,000 × g for 15 minutes (Centra-CL2, Needham Heights, MA, USA) The same amount of 0.85% sterile physiological saline was added and the microbial cells were washed twice to prepare a cell suspension.
실시예 1: 시트러스 과일즙 및 에센셜 오일 처리군 제조Example 1: Citrus fruit juice and essential oil treatment group production
시트러스 과일즙(라임, 레몬, 깔라만시)을 이용하여 10, 20%의 과일즙 용액을 제조하였다. 98% 에탄올을 이용하여 카바크롤, 티몰의 100, 150, 200 mM 표준 용액을 각각 제조하였다. 멸균된 튜브에 각 과일즙 용액을 9.8 ml 씩 담고 각 처리군에 해당하는 농도의 에센셜 오일 표준 용액 0.1 ml을 첨가하고 균질화하여 최종적으로 1.0, 1.5, 2.0 mM의 에센셜 오일을 함유한 과일즙 용액을 제조하였다. 제조를 마친 용액은 22°C로 설정된 교반항온수조에서 예열하였다. 에센셜 오일을 첨가하지 않은 과일즙과 멸균 증류수로 제조한 에센셜 오일 용액을 대조군으로 사용하였다. 각 1.0, 1.5, 2.0 mM 에센셜 오일의 %함량은 표 3과 같다.Citrus fruit juice (lime, lemon, calamanshi) was used to prepare 10, 20% fruit juice solution. 98% ethanol was used to prepare 100, 150 and 200 mM standard solutions of carbacrol and thymol, respectively. To each sterilized tube, add 9.8 ml of each fruit juice solution, add 0.1 ml of the essential oil standard solution corresponding to each treatment group, and homogenize the fruit juice solution containing 1.0, 1.5 and 2.0 mM of essential oil. . The prepared solution was preheated in a stirred constant temperature water bath set at 22 ° C. Fruit juice without essential oil and essential oil solution prepared with sterile distilled water were used as a control. The contents of 1.0, 1.5, and 2.0 mM essential oils are shown in Table 3.
실험예Experimental Example : : 시트러스Citrus 과일즙과 Fruit juice 에센셜Essential 오일 조합처리 항균력 시너지 효과 확인 Confirm synergy of antimicrobial effect of oil combination treatment
과일즙의 항균력 시너지 효과를 확인하기 위해 상기방법에 따라 제조한 에센셜 오일 함유 시트러스 과일즙(에센셜 오일: 1, 1.5, 2 mM / 과일즙: 10, 20%)에 6.5-7.5 log CFU/ml 수준의 균액을 접종하고 22°C 교반항온수조에서 5분 처리 후 균액을 증균배지에 도말하였다. 에센셜 오일을 첨가하지 않은 과일즙과 멸균 증류수로 제조한 에센셜 오일 용액에 대해서도 대조군으로서 같은 실험을 수행하였다. 도말한 배지는 37°C에서 24시간 배양한 후 배지 상에 형성된 콜로니를 계수하여 각 조건별 균 저감화 값을 log 값으로 산출하였다. 모든 실험은 3반복 수행되었으며 그 결과는 도 1-12와 같다(도 1: 일반 배양 E. coli O157:H7 - 카바크롤 함유 과일즙 용액 / 도 2: 일반 배양 E. coli O157:H7 - 티몰 함유 과일즙 용액 / 도 3: 산적응 배양 E. coli O157:H7 - 카바크롤 함유 과일즙 용액 / 도 4: 산적응 배양 E. coli O157:H7 - 티몰 함유 과일즙 용액 / 도 5: 일반 배양 S. Typhimurium - 카바크롤 함유 과일즙 용액 / 도 6: 일반 배양 S. Typhimurium - 티몰 함유 과일즙 용액 / 도 7: 산적응 배양 S. Typhimurium - 카바크롤 함유 과일즙 용액 / 도 8: 산적응 배양 S. Typhimurium - 티몰 함유 과일즙 용액 / 도 9: 일반 배양 L. monocytogenes - 카바크롤 함유 과일즙 용액 / 도 10: 일반 배양 L. monocytogenes - 티몰 함유 과일즙 용액 / 도 11: 산적응 배양 L. monocytogenes - 카바크롤 함유 과일즙 용액 / 도 12: 산적응 배양 L. monocytogenes - 티몰 함유 과일즙 용액).To confirm the synergistic effect of the fruit juice, 6.5-7.5 log CFU / ml was added to the citrus fruit juice (essential oil: 1, 1.5, 2 mM / fruit juice: 10, 20% And the broth was inoculated into the broth at 22 ° C for 5 minutes in a constant temperature water bath. The same experiment was carried out as a control for the fruit juice not containing the essential oil and the essential oil solution prepared with the sterilized distilled water. The cultured medium was incubated at 37 ° C for 24 hours, and colonies formed on the medium were counted. All experiments were repeated three times and the results are shown in Fig. 1-12 (Fig. 1: Normal culture E. coli O157: H7-carbacol fruit juice solution / Figure 2: Normal culture E. coli O157: fruit juice solution / 3: acid adaptive culture E. coli O157: H7 - cover scroll containing fruit juice solution / 4: adaptation acid culture E. coli O157: H7 - thymol-containing fruit juice solution / 5: General culture S. Typhimurium - cover scroll containing fruit juice solution / 6: General culture S Typhimurium - thymol-containing fruit juice solution / 7: San adaptation culture S Typhimurium - cover scroll containing fruit juice solution / 8: acid adaptive culture S Typhimurium - thymol-containing fruit juice solution / 9: General culture L. monocytogenes-carbazol scroll containing fruit juice solution / 10: General culture L. monocytogenes - thymol-containing fruit juice solution / 11: L. monocytogenes culture adaptation acid-carbazole crawl Containing fruit juice solution / Figure 12: acid-adapted culture L. monocytogenes -thymol-containing fruit juice solution) .
실험결과, 일반 배양 E. coli O157:H7와 산적응 배양 E. coli O157:H7 모두 (도 1-4) 2 mM의 카바크롤 또는 티몰을 과일즙(10, 20%)과 함께 처리한 경우 22°C에서 5분이라는 단시간 처리만으로도 큰 항균력 시너지 효과가 나타나 모든 균이 사멸하는 것으로 밝혀졌다. 1.5 mM의 카바크롤과 티몰을 처리하는 것만으로도 높은 수준의 항균력 시너지 효과가 나타났지만 모든 균을 저해하지는 못하였다.As a result of the experiment, when 2 mg of carbacol or thymol was treated with fruit juice (10, 20%) in both normal cultured E. coli O157: H7 and acid-adapted cultured E. coli O157: A short time treatment of 5 minutes at < RTI ID = 0.0 > C < / RTI > Treatment of 1.5 mM carbacrol and thymol showed synergistic effects at a high level but did not inhibit all bacteria.
S. Typhimurium과 L. monocytogenes의 경우에는 항균물질에 대해 E. coli O157:H7보다 낮은 저항성을 보이기 때문에 일반 배양균과 산적응 배양균 모두에서 E. coli O157:H7보다 훨씬 높은 저감화 값이 나타났다. 한 경우(산적응 배양 L. monocytogenes)만을 제외하고는 1.5 mM의 카바크롤 또는 티몰을 과일즙(10, 20%)과 함께 처리한 경우 22°C에서 5분이라는 단시간 처리만으로 모든 균을 사멸시킬 수 있었다. 1 mM의 카바크롤과 티몰을 처리하는 것만으로도 높은 수준의 항균 시너지 효과를 관찰할 수 있었지만 모든 균을 저해하지는 못하였다. S. In the case of Typhimurium and L. monocytogenes , E. coli O157: H7, it was found that E. coli O157: H7. In the case of treatment with 1.5 mM carbacol or thymol with fruit juice (10, 20%) except for one case (acid-adapted culture L. monocytogenes ), all the bacteria are killed only by a short treatment of 5 minutes at 22 ° C I could. A high level of antimicrobial synergy was observed by treatment with 1 mM carbacrol and thymol, but not all bacteria were inhibited.
산적응 배양균의 경우 일반 배양균보다 내산성이 강하므로 같은 농도의 에센셜 오일과 과일즙 혼합용액에 처리하였을 때 비교적 낮은 저감화 값을 나타냈지만, 깔라만시즙의 경우에는 큰 차이가 나타나지 않거나 오히려 더 높은 저감화 값을 나타내기도 하였다.Acid-adapted cultivars were more acid resistant than normal cultures, so they showed relatively low reduction values when treated with the same concentration of essential oil and fruit juice mixture. However, in the case of Kalaman juice, no significant difference or higher And a reduction value was also shown.
시트러스 과일즙(라임, 레몬, 깔라만시) 간의 시너지 효과 비교에서는 깔라만시, 레몬, 라임 순으로 에센셜 오일과 조합처리 시 큰 항균력 시너지 효과가 나타났다.In the comparison of synergy between citrus fruit juice (lime, lemon and calamanshi), there was a synergistic effect of large antibacterial activity in combination with essential oils in order of calamanshi, lemon and lime.
실험예Experimental Example 2: 2: 시트러스Citrus 과일즙 외 유기산과 Fruit juice and other organic acids 에센셜Essential 오일 조합처리 항균력 비교 Comparison of antimicrobial activity of oil combination treatment
실험결과 시트러스 과일즙 간의 시너지 효과가 깔라만시, 레몬, 라임 순으로 높게 나타났는데, 이 과일즙들은 동일한 농도에서 동일한 pH를 가졌다(10%: pH 2.6, 20%: pH 2.5). 따라서 시트러스 과일즙과 에센셜 오일간의 항균력 시너지 효과가 단순히 pH에 의한 작용이 아니라는 점에 착안하여 유기산을 이용하여 보충 실험을 진행하였다.The results showed that the synergistic effects of Citrus fruit juice were higher in order of Calamansi, Lemon and Lime. The fruit juices had the same pH at the same concentration (10%: pH 2.6, 20%: pH 2.5). Therefore, we focused on the fact that the synergy effect between citrus fruit juice and essential oil is not simply due to pH, and supplemental experiment was conducted using organic acid.
비교 대상 유기산으로는 일반적으로 과일즙에 많이 함유되어있는 구연산(citric acid, CTA)과 pH 적정 시 사용하는 염산(hydrochloric acid, HCl)을 선정하였고 20% 시트러스 과일즙과 동일한 pH인 2.5로 맞추어 실험을 진행하였다. 산적응 배양균으로 실험을 진행하였고, 에센셜 오일(카바크롤, 티몰)은 과일즙 간의 차이가 크게 나타난 1.5 mM을 사용하였다. 처리온도와 처리시간은 기존과 동일한 22°C, 5분으로 진행하였다. 모든 실험은 3반복 수행되었으며 그 결과는 도 13-14와 같다(도 13: 산적응 배양 E. coli O157:H7 - 카바크롤 함유 유기산, 과일즙 용액 / 도 14: 산적응 배양 E. coli O157:H7 - 티몰 함유 유기산, 과일즙 용액).Citric acid (CTA) and hydrochloric acid (HCl) used for pH titration were selected as the organic acids to be compared. The pH was adjusted to 2.5, which is the same pH as 20% citrus fruit juice . Experiments were performed with acid - adapted cultures. Essential oils (carbacol, thymol) were used at 1.5 mM, which showed a significant difference between fruit juices. The treatment temperature and treatment time were the same as the conventional one at 22 ° C for 5 minutes. All experiments were performed three replications The results are shown in Figure 13-14 (Fig. 13: Adaptation acid culture E. coli O157: H7 - cover crawl-containing organic acids, fruit juice solution / 14: acid adaptive culture E. coli O157: H7-thymol containing organic acid, fruit juice solution).
실험결과, 같은 pH 수준(pH 2.5)에서 유기산(구연산, 염산)을 에센셜 오일과 조합처리 했을 때보다, 과일즙(레몬, 깔라만시)을 에센셜 오일과 조합처리 한 경우 훨씬 높은 수준의 항균력 시너지 효과가 나타났다. 이를 통해 과일즙과 에센셜 오일의 조합처리 시 관찰되는 높은 수준의 항균력 시너지 효과는 단순히 pH에 의한 것이 아니라는 것을 확실시 할 수 있었고 특별한 기작이 있다는 점을 유추해볼 수 있었다.Experimental results show that the combination of fruit juice (lemon, calamanshi) with essential oils, compared with the combination of organic acids (citric acid, hydrochloric acid) and essential oils at the same pH level (pH 2.5) Effect. It can be concluded that the synergistic effects of high levels of antimicrobial activity observed in the combination of fruit juice and essential oil are not due to pH, and there is a special mechanism.
Claims (10)
Fruit juice from one or more fruits selected from calamansi, lemon and lime; And one or more essential oils selected from carbacol and thymol are mixed and applied to food to kill microorganisms.
The method of claim 1, wherein the microorganism is a food poisoning microorganism.
The method according to claim 1, wherein the microorganism is chromotherapy tumefaciens Vejle kyum (C. violaceum), Pseudomonas her labor Rouge (P. aeruginosa), Escherichia coli (E. coli), wave stew Pasteurella H. Molina urticae (P. hemolytica), Staphylococcus aureus (S. aureus), salmonella cola rasu devices (S. cholerasuis). L. monocytogenes jeneseu (L. monocytogenes), Salmonella typhimurium (S. Typhimurium), Campylobacter Jeju Needle (Campylobacter jejuni , Yersinia enterocolitica ), Cronobacter spp.), Shigella (Shigella spp.), Vibrio bacteria (Vibrio spp.), Clostridium perfringens (Clostridium botulinum ) and Bacillus spp.).
상기 과일즙은 1 내지 20%의 농도인, 방법.
The method according to claim 1,
Wherein the fruit juice has a concentration of 1 to 20%.
상기 과일즙 및 상기 에센셜오일을 혼합하여 제조되는 용액의 에센셜 오일의 최종 농도가 1 내지 2 mM인, 방법.
The method according to claim 1,
Wherein the final concentration of the essential oil in the solution prepared by mixing the fruit juice and the essential oil is 1 to 2 mM.
Fruit juice from one or more fruits selected from calamansi, lemon and lime; And at least one essential oil selected from carbacrol and thymol.
The composition according to claim 6, wherein the microorganism is a food poisoning microorganism.
The method according to claim 6, wherein the microorganism is chromotherapy tumefaciens Vejle kyum (C. violaceum), Pseudomonas her labor Rouge (P. aeruginosa), Escherichia coli (E. coli), wave stew Pasteurella H. Molina urticae (P. hemolytica), Staphylococcus aureus (S. aureus), salmonella cola rasu devices (S. cholerasuis). L. monocytogenes jeneseu (L. monocytogenes), Salmonella typhimurium (S. Typhimurium), Campylobacter Jeju Needle (Campylobacter jejuni , Yersinia enterocolitica ), Cronobacter spp.), Shigella (Shigella spp.), Vibrio bacteria (Vibrio wherein the composition is at least one selected from the group consisting of Clostridium perfringens , Clostridium botulinum and Bacillus spp.
상기 과일즙은 1 내지 20%의 농도인, 조성물.
The method of claim 6,
Wherein the fruit juice has a concentration of 1 to 20%.
상기 과일즙 및 상기 에센셜오일을 혼합하여 제조되는 용액의 에센셜 오일의 최종 농도가 1 내지 2 mM인, 조성물.
The method of claim 6,
Wherein the final concentration of the essential oil in the solution prepared by mixing the fruit juice and the essential oil is 1 to 2 mM.
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