KR100917802B1 - A composition using antibacterial and antifunga containing Thujopsis dolobrata oil - Google Patents

Abstract

The present invention relates to an essential oil component of the plant of the genus Thujopsis having antibacterial and antifungal activity and a composition comprising the same. More specifically, the present invention relates to an essential oil extract of the plant of the genus Thujopsis sp. Having antibacterial and antifungal activity and an antimicrobial and antifungal composition comprising the extract.

Nahanbaek essential oil according to the present invention has antimicrobial activity against skin diseases, purulent diseases, surgical wound infections, urinary tract-inducing bacteria and food poisoning bacteria, acne bacteria, and athlete's foot, so skin infections caused by pathogenic bacteria, purulent, otitis media, cystitis, It can be very useful for the prevention or treatment of athlete's foot by purulent diseases such as peritonitis, urinary tract infection, food poisoning, acne, staphylococcal infection, bacterial dysentery and pathogenic fungi. In addition, Nahanbaek essential oil extract of the present invention can be prepared in the form of cosmetics, disinfectants, sterile detergents and food preservatives.

Description

A composition using antibacterial and antifunga containing Thujopsis dolobrata oil}

The present invention relates to an antibacterial and antifungal composition comprising Nahanbaek essential oil, and more particularly, to an essential oil extract of Nahubaek (Thujopsis sp.) Plant having antibacterial and antifungal activity and an antibacterial and antifungal composition comprising the extract. will be.

Thujopsis dolabrata is classified as the Thujopsis of the Cupressaceae family, which is inhabited by moderate and fertile sandy soils in southern Japan. The Akajiwa Recreation Forest in the Kagura-gun is called Japan's three most beautiful forests along with cedars in Akida prefecture and hiba trees in Aomori prefecture.

The leaves of Nahanbaek are dark green with gloss, with white pore ships on the back, and the ones on the side spread obliquely, flat to the left and right, 4-6mm long in the shape of half, and the front and rear are flat and wide inverted egg shape. The end is dull. The leaves are vinyl-shaped, thicker and larger than cypress, and the surface of the leaves is serrated, making it easy to distinguish them. The leaves face each other and the back of the leaf is white except for the edge.

The pine cones, the fruit of Nahanbaek, ripen in October and have 1 to 1.5cm in diameter, and have 8 to 10 threaded pieces, and when they ripen, the threaded pieces are different in shape, but the ends are all like triangular awls. There is. Seeds 3 to 5 in each piece, with wings on each side, 4 to 5 mm long with wings, and butterflies 3 to 3.5 mm, oval shaped.

In addition, the flowers of Nahanbaek bloom in April-May, one at the end of the branches, one male and female, its stem is 10-30m in height, 90cm in diameter, the bark is grayish brown, and it is peeled off vertically and shallowly. The branches are shifted and flat, and the butterflies of the branches are 4 to 6 mm.

The inventor of the present applicant belongs to the Nahanbaek having the characteristics as described above for many years to isolate the new antimicrobial material from natural products, the present invention by confirming that the genus plants have antibacterial activity against various pathogenic bacteria and fungi Completed.

The present invention has been made to solve the above problems, the purpose of the skin disease, purulent diseases, surgical wound infection, urinary tract infections using essential oil components of the plant of the genus Thujopsis having antibacterial and antifungal activity It has antibacterial activity against bacteria and food poisoning bacteria, acne bacteria, and athlete's foot fungus, so it is purulent diseases such as skin infection, purulent, otitis media, cystitis, peritonitis, urinary tract infection, food poisoning, acne, staphylococcal infection, bacterial dysentery and pathogenicity The present invention provides an antimicrobial and antifungal composition comprising Nahanbaek essential oil having antibacterial and antifungal activity which is very useful for preventing or treating athlete's foot.

According to a feature of the present invention for achieving the above object, in the antifungal composition containing Najobaek essential oil (Thujopsis dolobrata OIL), the genus Aspergillus sp and Trichophyton against the fungus in the composition sp.) or for yeast, a composition exhibiting antifungal activity against Candida sp.

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In addition, according to an additional feature of the present invention, it is preferable that the fungus of Trichophyton genus Tricophyton mentagrophytes or the fungus of Aspergillus genus is composed of Aspergillus niger.

In addition, according to an additional feature of the invention, it is preferred that the genus of Candida consists of Candida albicans.

Antimicrobial and antifungal compositions comprising the Thujopsis dolobrata oil according to the present invention have antimicrobial activity against skin diseases, purulent diseases, surgical wound infections, urinary tract infection-inducing bacteria and food poisoning bacteria, acne bacteria, and athlete's foot bacteria. It can be very useful for the prevention or treatment of athlete's foot by skin infection, purulent, otitis media, cystitis, peritonitis, purulent diseases, urinary tract infection, food poisoning, acne, staphylococcal infection, bacterial dysentery and pathogenic fungi.

In addition, Nahanbaek essential oil extract of the present invention can be prepared in the form of cosmetics, disinfectants, sterile detergents and food preservatives.

In order to achieve the above object, the present invention provides an extract of Nahanbaek essential oil having antibacterial and antifungal activity.

The present invention also provides an antimicrobial composition comprising the extract.

The present invention also provides an antifungal composition comprising the extract.

Nahanbaek essential oil as described above (Thujopsis dolobrata OIL) has antimicrobial activity against various pathogenic bacteria and antibacterial and antifungal activity against fungi, yeast.

The composition of Nahanbaek essential oil of the present invention is the genus Escherichia sp., Staphylococcus sp., Streptococcus sp., Bacillus sp., Bacillus sp., Pseudomonas sp. Pseudomonas sp.) Or Listeria sp., Which has antimicrobial activity against the bacteria selected from the group, Aspergillus sp. (Aspergillus sp) and Trichophyton sp. It has the antifungal activity of Candida sp. And Pityrosporum sp.

The Escherichia sp. Is commonly referred to as Escherichia coli, and is a common basic gram-negative bacillus with parasites of mammalian intestines, including humans. Is known to cause cystitis, pyelitis, peritonitis, sepsis and the like.

The Staphylococcus aureus is one of the bacteria that is widely distributed in the natural world, invading the skin, respiratory organs and genitourinary system, and is one of the representative causes of purulent diseases such as purulent, otitis media and cystitis as well as food poisoning. In particular, the bacteria are often infected with people with weak immunity, such as surgical patients, chronic bronchitis, cold patients. In particular, methicillin resistant Staphylococcus aureus is a strain having a high or high level of resistance due to loss of sensitivity to methicillin.

The Staphylococcus epidermidis is a non-pathogenic bacterium that is commonly present in the skin or the nose. However, in the case of supplying fluid through a vein, blood is collected, parasitises in the catheter used in the case of a surgical patient, causing infection of the wound, skin tissue infection and peritonitis, and in the Staphylococcus spp. Intermedius is known to cause cystitis or purulent dermatitis by infecting the skin or urinary tract of animals, especially dogs, and saprophyticus is known as the main cause of urinary tract infections in normal bacteria in women and in women.

In addition, Streptococcus mutans is a major causative agent of tooth decay, which breaks down sugar and converts it into lactic acid while inhabiting the oral cavity, causing erosion of the tooth, which is the main ingredient of lime, and inhibiting the activity of intestinal rot in our bodies. It reduces carcinogens and carcinogens such as ammonia and indole. It also absorbs these harmful substances and releases them out of the body.

In addition, anthracis in Bacillus is the causative agent of anthrax, a common infectious disease. The anthrax is infected with most vertebrates and is known to have a high incidence of cattle or goats. In humans, most are infected when exposed to infected animals or their by-products.

Pseudomonas aeruginosa is a green-growing bacterium that is not life-threatening and is generally known to be infected by piercing.

Listeria monocytogenes is a food poisoning bacterium that causes food poisoning. Phytophyton mentagrophytes are skin fungi that cause skin fungal infections such as athlete's foot, and fungal infections, such as Candida albicans, which cause itching on the folds of the neck or groin. It is known to produce red papules or pustules.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings.

Figure 1 is a graph analyzing the components of Nahanbaek essential oil using an analyzer, Figure 2 shows an exemplary diagram indicating a specific component in the results according to Figure 1, Figures 3a to 3h is Nahanbaek essential oil of antibacterial efficacy It is an experimental example showing the activity of the clear zone (clear zone), Figure 4a to 4d shows an experimental example showing the activity of the inhibitory growth of Nahanbaek essential oil antifungal effect.

Nahanbaek essential oil of the present invention as shown in the configuration of the experimental example shows the characteristics of inhibiting the activity of the antibacterial and antifungal effect against various bacteria, fungi or yeast, for the antibacterial and antifungal composition of Nahanbaek essential oil according to the present invention Looking at the characteristics through an embodiment, first, the components of the Nahanbaek essential oil of the present invention is configured as follows.

Example

Experimental Example 1.

GC / MS analysis was performed to determine the components of the Nahanbaek essential oil, and the conditions are as follows.

A model HP 6890 GC instrument was used for GC analysis and a column of HP-5 (length 30 m, diameter 0.25 mm) was used. The oven temperature was maintained at 50 ° C. for 3 minutes, and then heated up by 5 ° C./min to 350 ° C. for the final temp.

MS used HP 5973 MSD and analyzed the components of Nahanbaek essential oil according to the present invention in EI mode.

Table 1

Nahanbaek essential oil ingredient list

No Rt (min) % of total Components Formula / Mw One 12.87 0.37  α-Terpinolene C10H16 /136.23 2 15.83 1.08  3-cyclohexen-1-ol C10H18O / 154.25 3 16.24 0.17  α-Terpineol (p-menth-1-en-8-ol) C10H18O / 154.25 4 17.12 0.29  β-Citronellol C10H20O / 156.27 5 19.16 0.77  Carvacrol C10H14O / 150.22 6 19.75 0.84  Dicyclopropyl ketone (C3H5) 2CO / 110.15 8 21.11 0.10  Anethole C10H12O / 148.20 9 21.65 1.20  β-Elemene C15H24 /204.35 10 21.79 0.27  Isolongifolene C15H24 /204.35 11 22.03 0.65  α-Longipinene C15H24 /204.35 12 22.20 0.09  3-Propyltropolone 13 22.38 1.45  α-Cedrene C15H24 /204.35 14 22.61 0.77  γ-Muurolene C15H24 /204.35 15 23.07 31.89  Thujopsene C15H24 /204.35 16 23.24 0.18  β-Farnesene C15H24 /204.35 17 23.31 0.26  α-Farnesene C15H24 /204.35 18 23.38 0.27  α-Farnesene C15H24 /204.35 19 23.69 0.27  9,10-Dehydro-Isolongifolene C15H22 /202.33 20 23.79 0.28  Epizonarene C15H24 /204.35 21 23.91 1.42  β-Thujaplicin C10H12O2 /164.20 23 24.06 1.62  β-Chamigrene C15H24 /204.35 24 24.22 0.40  β-Eudesmene C15H24 /204.35 25 24.40 0.43  α-Selinene C15H24 /204.35 26 24.62 5.50  β-Himachalene C15H24 /204.35 27 24.72 2.55  (+)-Cuparene C15H22 /202.34 28 24.91 1.24  δ-Cadinene C15H24 /204.35 29 25.04 0.87  β-Sesquiphellandrene C15H24 /204.35 30 25.35 3.14  β-himachalene C15H24 /204.35 31 25.66 1.11  Cyclohexane, 1,3,5-trimethyl-, (1,3,5) -cis C9H18 /126.24 32 26.60 0.21  Bicyclogermacrene C15H24 /204.35 33 26.85 1.33  β-Bisabolene C15H24 /204.35 34 27.20 8.62  Cedrol C15H26O / 222.37 35 27.31 0.15  α- Himachalene C15H24 /204.35 36 27.54 0.59  Cadina-1,4-diene C15H24 /204.35 37 27.65 0.44  γ-Curcumene C15H22 /202.33 38 27.76 0.58  α-Cedrene C15H24 /204.35 39 27.88 0.50  τ-Cadinol C15H26O / 222.37 40 28.24 2.57  Aromandendrene C15H24 /204.35 41 28.38 1.77  α-Longipinene C15H24 /204.35 42 28.87 5.68  Germacrene b C15H24 /204.35 43 31.08 0.37  Camphene C10H16 / 136.23 44 31.18 0.40  Valerenal C15H22O / 218.33 45 31.62 0.20  Cuminaldehyde C10H12O / 148.20 46 17.11  unknown

As shown in Table 1 above, the components of the Nahanbaek essential oil are analyzed and labeled graphically as shown in FIG. 1, and as shown in FIG. 2, according to the present invention. It can be seen that the Thujopsene ingredient of the one hundred essential oils contains the most.

Experimental Example 2.

Strain Culture Conditions

For each strain, the medium (BRC-KCTC) purchased from Korea Biotechnology Center or ATCC was used. All mediums were sterilized at 121 ° C. for 15 minutes and then injected into a petri dish to perform experiments with a solid plate medium. The liquid medium was broth of the same composition without agar powder.

The cells were lyophilized and streaking in a solid medium using a holding loop, followed by incubation at a specified temperature and incubation days. After incubation, a single colony was inoculated into the liquid medium. After inoculation into the liquid medium and cultured in a shaking incubator at 37 ℃, the growth of the strain was measured by absorbance at 600 nm using a UV / VIS spectrophotometer, using the paper disc method using the strain of the best time zone Antimicrobial activity was measured, and the turbidity was measured by adjusting the inoculation amount so that the cultured cells were 2 to 3 X 10 ⁶ / mL and 2 to 3 X 10 ⁷ / mL, as shown in Table 2. For the antimicrobial activity test, two representative E. coli strains (E. coli: KCTC2571, KCTC2593), Bacillus subtilis (B. subtilis: KCTC2189), and Listeria monocytos among Listeria bacteria were tested. L. monocytogenes (KCTC3710), Staphylococcus aureus (S. aureus: ATCC6538P) or Staphylococcus epidermidis (KCTC1917), S. mutans (KCTC3065) among Leptococcus bacteria and P. aeruginosa (KCTC2513) among Pseudomonas bacteria were cultured, and trichophyton spp. Against fungi for antifungal activity test T. mentagrophytes (KCTC6316) in the genus Aspergillus and A. niger (KCTC6317) in the genus Aspergillus and Candida albicans (KCTC7546) in the Candida genus for yeast Bacteria, P. tyrosporum obare (P.ovale) in the genus Pytylosporum.

TABLE 2

Strains Used and Culture Conditions

Strain name Strain number Classification Culture medium (KCTC #) Growth temperature (℃) Remarks One Esherichia coli (E. coli) KCTC2571 bacteria (aerobic) One 37 2 Esherichia coli KCTC2593 bacteria (aerobic) One 37 3 Basillus subtilis Bacillus subtilis KCTC2189 bacteria (aerobic) One 30 4 Listeria monocytogenes (Listeria) KCTC3710 bacteria (aerobic) 3 37 5 Staphylococcus aureus (Staphylococcus aureus) ATCC6538P bacteria (aerobic) 1/20/98 37  ATCC Badge 6 Pseudomonas aeruginosa Pseudomonas aeruginosa KCTC2513 bacteria (aerobic) One 37 7 Streptococcus mutans KCTC3065 bacteria (anaerobic) 3 37 8 Streptococcus epidermidis (Skin Staphylococcus) KCTC1917 bacteria (aerobic) One 37 9 Trichophyto mentagrophytes Athlete's foot KCTC6316 mold (aerobic) 340 28 10 Aspergillus nigers (Black mold) KCTC6317 mold (aerobic) 31 24 11 Candida albicans KCTC7965 yeast (aerobic) 26 25 12 Pityrosporum ovale (M. furfur ) (Dandruff) KCTC7546 yeast (aerobic) 429 30

Experimental Example 3.

Antifungal Activity Measurement

A single colony of each strain cultured above is taken and seeded in 2 ml sterile liquid medium. After incubation by incubation in a shaking incubator or incubator for a specified temperature and incubation time, the culture medium is diluted to adjust the bacteria to absorbance at 600 nm. Take 200 ~ 300 ㎕ of the liquid medium and spread it evenly with a spreader on the solid medium, and place a sterile 6 mm filter paper disc on the plate. 5 μl of each essential oil sample is injected into the filter paper disc, inhaled and placed on a clean bench, followed by incubation at 37 ° C. (or 25, 30 ° C.) for 24 to 96 hours depending on the incubation time of each strain. After incubation, antifungal activity was confirmed by measuring the diameter (mm) of the clear zone (CZ) around the filter paper disc.

TABLE 3

Antibacterial and Antifungal Activity of Nahanbaek Essential Oil against Various Strains

Test strain Gram dyeing Strain number Clear Zone (mm) Remarks One E.coli - KCTC2571 23.3 ± 1.5 crude 2 E.coli - KCTC2593 25.0 ± 2.0 crude 3 B.subtillis + KCTC2189 29.7 ± 2.9 crude 4 L.monocytogenes + KCTC3710 31.0 ± 2.0 crude 5 S.aureus + KCTC6538P 23.3 ± 3.2 crude 6 P.aeruginosa - KCTC2513  9.3 ± 0.6 crude 7 S.mutans + KCTC3065 31.5 ± 6.4 crude 8 S.epidermidis + KCTC1917 28.5 ± 2.1 crude 9 T.mentagrophytes KCTC6316 39.0 ± 8.5 1/25 crude 10 A.nigers KCTC6317 26.7 ± 1.5 crude 11 C.albicans KCTC7965 41.7 ± 2.1 crude 12 P.ovale (M.furfur) KCTC7546 13.3 ± 0.6 crude

According to the above experiments, the antibacterial activity of each strain was inhibited in Nahanbaek essential oil as shown in FIGS. 3A to 3H attached thereto, and the antifungal activity of Nahanbaek essential oil against fungi as shown in FIGS. 4A to 4B. The antifungal activity of Nahanbaek essential oil was inhibited against yeast as shown in Figs. 4C to 4D. (White arrow)

The present invention is not limited to the embodiments described above, and various modifications and changes can be made by those skilled in the art, which are included in the spirit and scope of the present invention as defined in the appended claims.

1 is a graph analyzing the components of Nahanbaek essential oil using an analyzer

2 is an exemplary diagram showing a specific component in the result according to FIG.

Figure 3a is a picture showing the activity of the clear zone of the antibacterial effect of Nahanbaek essential oil according to the present invention Escherichia coli (E. colii; KCTC2571).

Figure 3b is a picture showing the activity of the clear zone of the antibacterial effect of Nahanbaek essential oil according to the present invention Escherichia coli (E. colii; KCTC2593).

Figure 3c is a picture showing the activity of the clear zone of the antibacterial effect of Nahanbaek essential oil according to the present invention Bacillus subtilis (B. subtilis; KCTC2189) in the genus Bacillus.

Figure 3d is a picture showing the activity of the clear zone of the antibacterial effect of Nahanbaek essential oil according to the present invention Listeria monocytogenes (L. monocytogenes; KCTC3710) of the genus Listeria.

Figure 3e is a photograph showing the activity of the clear zone of the antibacterial effect of Nahanbaek essential oil according to the present invention Staphylococcus aureus (S. aureus; ATCC6538P) in the genus Staphylococcus.

Figure 3f is a photograph showing the activity of the clear zone of the antibacterial effect of P. aeruginosa (P. aeruginosa; KCTC2513) in the genus Pseudomonas Nahanbaek essential oil according to the present invention.

Figure 3g is a picture showing the activity of the clear zone of the antibacterial effect of Nahanbaek essential oil according to the present invention Streptococcus mutans (S. mutans; KCTC3065) of the genus Streptococcus.

Figure 3h Nahanbaek essential oil according to the present invention Staphylococcus epidermidis (S. epidermidis; KCTC1917) of the genus Staphylococcus photos showing the activity of the antimicrobial effect (clear zone) growth.

Figure 4a is a picture showing the activity of the clear zone of the antifungal effect of Nahanbaek essential oil according to the present invention trichophyton Mentagrophytes (T. mentagrophytes; KCTC6316) in the fungus of the genus Tricophyton.

Figure 4b is a picture showing the activity of the clear zone of the antifungal effect of Nahanbaek essential oil according to the present invention Aspergillus niger (A. nigers; KCTC6317) of the genus Aspergillus.

Figure 4c is a picture showing the activity of the clear zone of the antifungal efficacy of Nahanbaek essential oil according to the present invention Candida albicans (C. albicans; KCTC7965) of yeast genus Candida.

Figure 4d is a picture showing the activity of the clear zone of the antifungal effect of Nahanbaek essential oil according to the present invention Pyrosporum obare (P. ovale; KCTC7546) of yeast in the genus Pytirosporum.

Claims (12)

delete delete In the antifungal composition containing Nahubaek essential oil (Thujopsis dolobrata OIL), The composition characterized in that it exhibits antifungal activity against Aspergillus sp and Trichophyton sp. Against a fungus or Candida sp. Against a yeast. delete delete delete delete delete The method of claim 3, wherein The tricophyton fungi composition is characterized in that consisting of tricophyton mentagrophytes (Trichophyton mentagrophytes). The method of claim 3, wherein The composition of the genus Aspergillus is composed of Aspergillus niger (Aspergillus niger). The method of claim 3, wherein The composition of the genus Candida is composed of Candida albicans (Candida albicans) delete

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