• 한국유기농업학회지
• /
• 제23권4호
• /
• pp.887-896
• /
• 2015

본 연구는 균핵병의 원인균인 Sclerotinia sclerotiorum에 대해 항균활성을 갖는 친환경 유기농자재를 개발할 목적으로 송라(Usnea longissima)로부터 활성물질을 분리하였다. 송라를 MeOH로 추출하여 용매분획을 하였고, 용매분획 중에서 강한 활성을 나타낸 $CHCl_3$ fraction을 column chromatography로 분리하여 40개의 subfractions을 얻었고 9개의 그룹으로 나누었다. 9개의 subfractions을 bioassay한 결과 G3 subfraction에서 강한 활성을 나타내었다. G3 subfraction을 GC-MS로 분석하여 chromatogram 상의 활성물질로 추정되는 peak의 mass spectrum과 Wiley library를 비교하여 profiling한 결과, Usnic acid가 주요 물질로 검출되었다. 항균활성물질로 추정되는 Usnic acid 화합물의 항균활성을 검정하기 위하여 표준품을 사용하여 생물검정한 결과 이 화합물이 강한 활성을 나타내었다. 따라서 송라로부터 분리한 Usnic acid가 균핵병의 원인균인 Sclerotinia sclerotiorum에 대한 항균 활성물질인 것으로 추정하였다.

• Bulletin of the Korean Chemical Society
• /
• 제34권6호
• /
• pp.1684-1688
• /
• 2013

A rapid and sensitive method for determining usnic acid of Lethariella cladonioides in rat was established using high performance liquid chromatography (HPLC) quadrupole time-of-flight (QTOF) tandem mass (MS/MS). Rat plasma was pretreated by mixture of acetonitrile and chloroform to precipitate plasma proteins. Chromatographic separation was achieved on a column ($50{\times}2.1$ mm, $5{\mu}m$) with a mobile phase consisting of water (containing $5{\times}10^{-3}$ M ammonium formate, pH was adjusted to 3.0 with formic acid) and acetonitrile (20:80, v/v) at a flow rate of 0.3 mL/min. A tandem mass spectrometric detection with an electrospray ionization (ESI) interface was conducted via collision induced dissociation (CID) under negative ionization mode. The MS/MS transitions monitored were m/z 343.0448 ${\rightarrow}$ m/z 313.2017 for usnic acid and m/z 153.1024 ${\rightarrow}$ m/z 136.2136 for protocatechuic acid (internal standard). The linear range was calculated to be 2.0-160.0 ng/mL with a detection limit of 3.0 pg/mL. The inter- and intra-day accuracy and precision were within ${\pm}7.0%$. Pharmacokinetic study showed that the apartment of usnic acid in vivo confirmed to be a two compartment open model. The method was fully valid and will probably be an alternative for pharmacokinetic study of usnic acid.

• The Plant Pathology Journal
• /
• 제38권1호
• /
• pp.25-32
• /
• 2022

Plant pathogens pose major threats on agriculture and horticulture, causing significant economic loss worldwide. Due to the continuous and excessive use of synthetic pesticides, emergence of pesticide resistant pathogens has become more frequent. Thus, there is a growing needs for environmentally-friendly and selective antimicrobial agents with a novel mode of action, which may be used in combination with conventional pesticides to delay development of pesticide resistance. In this study, we evaluated the potentials of lichen substances as novel biopesticides against eight bacterial and twelve fungal plant pathogens that have historically caused significant phytopathological problems in South Korea. Eight lichen substances of diverse chemical origins were extracted from axenic culture or dried specimen, and further purified for comparative analysis of their antimicrobial properties. Usnic acid and vulpinic acid exhibited strong antibacterial activities against Clavibacter michiganensis subsp. michiganensis. In addition, usnic acid and vulpinic acid were highly effective in the growth inhibition of fungal pathogens, such as Diaporthe eres, D. actinidiae, and Sclerotinia sclerotiorum. Intriguingly, the growth of Rhizoctonia solani was specifically inhibited by lecanoric acid, indicating that lichen substances exhibit some degrees of selectivity to plant pathogens. These results suggested that lichen substance can be used as a selective biopesticide for controlling plant disease of agricultural and horticultural significance, minimizing possible emergence of pesticide resistant pathogens in fields.

• BMB Reports
• /
• 제34권3호
• /
• pp.194-200
• /
• 2001

Secreted arginase from Evernia prunastri thallus has been purified 616-fold from the incubation medium. Purified arginase was resolved as only one peak in a capillary electrophoresis with a pI value of 5.35. The protein contained high amounts of acidic amino acids, such as Asx and Glx, and a relatively high quantity of Ser and Gly. The molecular mass of native, purified arginase was estimated as about 26 kDa by SE-HPLC. Substrate saturated kinetic showed a typical Michaelis-Menten relationship with a K_m value of 3.3 mM L-arginine. Atranorin behaved as a mixed activator of the enzyme (apparent $K_m$ = 0.96 mM); whereas evernic and usnic acid were revealed as non competitive inhibitors (apparent $K_m$ values were 3.16 mM and 3.05 mM, respectively). Kinetics of atranorin binding indicated that saturation was reached from 0.18 ${\mu}mol$ of the total atranorin and the occurrence of multiple sites for the ligand. This agrees with a possible aggregation of several enzyme subunits during the interaction process. A value of binding sites of about 12 was obtained. The binding of evernic acid was saturated from 23 nmol of total phenol. The number of binding sites was about 5. The loss of the binding ability of evernic acid could be interpreted as a single negative cooperatively. Usnic acid behaves in a similar way to evernic acid, although the binding saturation occurs at $0.14\;{\mu}moles$ of the ligand. This binding appears to be unspecific, and has 28 usnic acid binding sites to the protein.

• Mycobiology
• /
• 제35권2호
• /
• pp.45-46
• /
• 2007

Lecanora muralis was found on the rock along coastal line during the field trip in Jeju island in 2006. Thallus crustose, placodioid, closely adnate, forming orbicular patches; upper surface grayish green, glossy; central lobes areolate, marginal parts plane, edges thin pruinose; lower surface ecorticate; apothecia sessile, lecanorine type, exciple dense and intact when young, and disc plane, but when mature, exciple laciniate, disc protrudent, yellowish brown to orange, $0.5{\sim}1.5$ mm in diameter; ascospores ellipsoid, simple, colorless, $12.5{\sim}15.0{\times}5.0{\sim}7.5\;{\mu}m$. Usnic acid and zeorin contained in thallus. This is the first record of this species in South Korea.

• 분석과학
• /
• 제36권2호
• /
• pp.89-98
• /
• 2023

Analytical methods for detecting atranol, chloroatranol, evernic acid, (+)-usnic acid, and atranorin in sanitary napkins and mouthwashes were developed using ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). UHPLC-MS/MS conditions were optimized for rapid, sensitive, and simultaneous analysis of the five allergenic compounds. The methods were validated by assessing their specificity, matrix effects, limit of detection (LOD), limit of quantification (LOQ), linearity, accuracy, and precision. Good linearity was achieved with a determination coefficient of ≥0.99. The LOD and LOQ were 2.1-9.8 and 6.4-29.6 ng/g for sanitary napkins and 0.29-0.48 and 0.87-1.45 ng/mL for mouthwashes, respectively. The accuracy and precision were within an acceptable range according to the criteria reported in the European SANTE/11813/2017 guidelines (70-120 % recovery, <20 % relative standard deviation). Therefore, these methods can be used to analyze atranol, chloroatranol, evernic acid, (+)-usnic acid, and atranorin in sanitary napkins and mouthwashes.

• 생약학회지
• /
• 제47권2호
• /
• pp.122-127
• /
• 2016

Methicillin-Resistant Staphylococcus aureus(MRSA) is a multidrug-resistant(MDR) strain. (+)-Usnic acid(UA) is uniquely found in lichens, and is especially abundant in genera such as Usnea and Cladonia. UA has antimicrobial activity against human and plant pathogens. Therefore, UA may be a good antibacterial drug candidate for clinical development. In search of a natural products capable of inhibiting this multidrug-resistant bacteria, we have investigated the antimicrobial activity of UA against 17 different strains of the bacterium. In this study, the effects of a combination of UA and permeable agents against MRSA were investigated. For the measurement of cell wall permeability, UA with concentration of Ethylenediaminetetraacetic acid(EDTA) was used. In the other hand, Sodium azide($NaN_3$) was used as inhibitors of ATPase. Against the 17 strains, the minimum inhibitory concentrations(MICs) of UA were in the range of $7.81-31.25{\mu}g/ml$. EDTA or $NaN_3$ cooperation against MRSA showed synergistic activity on cell wall. UA and in combination with EDTA and $NaN_3$ could lead to the development of new combination antibiotics against MRSA infection.

• 한국약용작물학회:학술대회논문집
• /
• 한국약용작물학회 2011년도 추계학술발표회
• /
• pp.356-357
• /
• 2011

• Mycobiology
• /
• 제34권3호
• /
• pp.148-150
• /
• 2006

Karoowia saxeti was recorded during the lichen field expedition in southern part of Korea in 2006. The lichen was found on the rock surface along coastal line. This species was easily recognized by chemistry (K+ yellow) and the presence of isidia. Thallus was saxicolous, subcrustose, more or less lobate at the center with clearly lobed margins, $2{\sim}6\;cm$ broad and pale yellowish green. Thalli lobes were irregular, variable, up to 1.0mm wide, not branched, flat to more or less convex and contiguous to subimbricate. Upper surface of the thalli was continuous, emaculate, moderately isidiate. The isidia was subglobose to cynlindrical, darkening at the tips and unbranched. Low surface of the thalii was black with a spongy rhizoidal and lamellar layer. HPLC analysis proved the presence of stictic acid (K+ yellow), norstictic acid and usnic acid. This is the first record of the species in South Korea.

• 한국미생물·생명공학회지
• /
• 제40권1호
• /
• pp.23-29
• /
• 2012

순천대학교 지의류생물 소재은행(KOLABIC)에서 100종의 지의류 내생 곰팡이를 분양 받아 C. albicans에 대한 항진균 활성을 조사하였다. 고체배지 및 액체배지에 배양시 Usnea rigidula, Parmotrema pseudotinctorum 그리고 Myelochroa sp.가 높은 항진균 활성을 나타내는 물질을 생성함을 알 수 있었다. 이들 3종 지의류 내생 곰팡이의 배양액을 chloroform, ethyl acetate 용매 순으로 추출하고 disc diffusion method를 이용하여 각 추출물의 C. albicans에 대한 항진균 활성도를 측정한 결과 모두 chloroform 추출물에서 높은 활성이 가장 높았으며, 다음으로 ethyl acetate 추출물이 높은 활성을 나타냈다. 항진균성 물질이 기존에 알려진 물질인지에 대한 정보를 알기위해 6 일 동안 배양한 Usnea rigidula와 4일 동안 배양한 Parmotrema pseudotinctorum의 chloroform 추출액에 대하여 HPLC, $C_{18}$ column으로 분리하여 Usnea rigidula (2326)의 6일과 Parmotrema pseudotinctorum (2202)의 4일 배양액에서 각각 2 개의 peak 분획물을 얻었다. 각 분획물을 농축한 후 96 well plate를 이용하여 C. albicans와 C. glabrata에 대한 항진균 활성을 실험한 결과, Parmotrema pseudotinctorum의 주요peak는 control에 비해 항진균 활성이 높게 나타났으며 활성물질은 usnic acid계열의 화합물로 추정되었다.