• The Plant Pathology Journal
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• 제38권2호
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• pp.115-130
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• 2022

Though information exists regarding the pathogenesis of the shot-hole disease (SH) in flowering cherry (FC), there has been a lack of research focusing on SH management. Therefore, here, we investigated the inhibitory activities of antagonistic bacteria against SH pathogens both in vitro and in vivo as well as their biochemical characteristics and bioactive compounds. Two biosurfactant-producing bacterial antagonists, identified as Bacillus velezensis strains JCK-1618 and JCK-1696, exhibited the best effects against the growth of both bacterial and fungal SH pathogens in vitro through their cell-free culture filtrates (CFCFs). These two strains also strongly inhibited the growth of the pathogens via the action of their antimicrobial diffusible compounds and antimicrobial volatile organic compounds (VOCs). Crude enzymes, solvent extracts, and biosurfactants of the two strains exhibited antimicrobial activities. Liquid chromatography/electrospray ionization time-of-flight mass spectrometric analysis of the partially purified active fractions revealed that the two antagonists produced three cyclic lipopeptides, including iturin A, fengycin A, and surfactin, and a polyketide, oxydifficidin. In a detached leaf assay, pre-treatment and co-treatment of FC leaves with the CFCFs led to a large reduction in the severity of the leaf spots caused by Epicoccum tobaicum and Bukholderia contaminans, respectively. In addition, the two antagonists produced indole-3-acetic acid, siderophore, and a series of hydrolytic enzymes, along with the formation of a substantial biofilm. To our knowledge, this is the first report of the antimicrobial activities of the diffusible compounds and VOCs of B. velezensis against the SH pathogens and their efficiency in the biocontrol of SH.

• 한국환경농학회지
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• 제41권1호
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• pp.50-54
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• 2022

BACKGROUND: Phyllospadix japonica Makino is a perennial plant belonging to the family Zosteraceae. This species is native to the eastern coast of the Korean Peninsula, and it is found attached to rocks on the seashore. As with all seagrass species, P. japonica is also known to play a major role in protecting the marine environment, and studies on its physiological activities suggestive of its antioxidant, antibacterial and anti-inflammatory potentials have been reported. In this study, purification and structural analysis were performed to identify the polyphenol substances derived from P. japonica. METHODS AND RESULTS: An polyphenol substance MP-1 was purified from the 70% aqueous methanol extract of P. japonica using Diaion® HP-20 column chromatography, ODS column chromatography, and medium-pressure liquid chromatography (MPLC). The purified MP-1 was identified as rosmarinic acid having a molecular weight of 360 and a molecular formula of C18H16O8 through electrospray ionization (ESI)-mass and nuclear magnetic resonance (NMR) spectroscopic analysis. CONCLUSION(S): This study highlights the processes used for the identification of the polyphenol substance derived from P. japonica. Rosmarinic acid, the polyphenol derived from P. japonica identified by this study, is a kind of bioactive substance mainly present in plants. These findings provide an important starting point and are valuable for future studies on bioactive substances in seagrass.

• Journal of Microbiology and Biotechnology
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• 제33권6호
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• pp.797-805
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• 2023

Species belonging to the Vernonia (Asteraceae), the largest genus in the tribe Vernonieae (consisting of about 1,000 species), are widely used in food and medicine. These plants are rich sources of bioactive sesquiterpene lactones and steroid saponins, likely including many as yet undiscovered chemical components. A phytochemical investigation resulted in the separation of three new stigmastane-type steroidal saponins (1 - 3), designated as vernogratiosides A-C, from whole plants of V. gratiosa. Their structures were elucidated based on infrared spectroscopy (IR), one-dimensional (1D) and two-dimensional nuclear magnetic resonance (2D NMR), high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), and electronic circular dichroism analyses (ECD), as well as chemical reactivity. Molecular docking analysis of representative saponins with α-glucosidase inhibitory activity was performed. Additionally, the intended substances were tested for their ability to inhibit α-glucosidase activity in a laboratory setting. The results suggested that stigmastane-type steroidal saponins from V. gratiosa are promising candidate antidiabetic agents.

• Animal Bioscience
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• 제36권9호
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• pp.1367-1375
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• 2023

Objective: Pigment production and distribution are controlled through multiple proteins, resulting in different coat color phenotypes of sheep. Methods: The expression distribution of vimentin (VIM) and transthyretin (TTR) in white and black sheep skins was detected by liquid chromatography-electrospray ionization tandem MS (LC-ESI-MS/MS), gene ontology (GO) statistics, immunohistochemistry, Western blot, and quantitative real time polymerase chain reaction (qRT-PCR) to evaluate their role in the coat color formation of sheep. Results: LC-ESI-MS/MS results showed VIM and TTR proteins in white and black skin tissues of sheep. Meanwhile, GO functional annotation analysis suggested that VIM and TTR proteins were mainly concentrated in cellular components and biological process, respectively. Further research confirmed that VIM and TTR proteins were expressed at significantly higher levels in black sheep skins than in white sheep skins by Western blot, respectively. Immunohistochemistry notably detected VIM and TTR in hair follicle, dermal papilla, and outer root sheath of white and black sheep skins. qRT-PCR results also revealed that the expression of VIM and TTR mRNAs was higher in black sheep skins than in white sheep skins. Conclusion: The expression of VIM and TTR were higher in black sheep skins than in white sheep skins and the transcription and translation were unanimous in this study. VIM and TTR proteins were expressed in hair follicles of white and black sheep skins. These results suggested that VIM and TTR were involved in the coat color formation of sheep.

• 분석과학
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• 제22권4호
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• pp.319-325
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• 2009

벌꿀 중에 잔류하는 아미노그라이코사이드 항생제인 네오마이신을 효과적으로 분석하는 방법을 개발하고 방법에 대한 유효성 검증을 수행하였다. 0.1M 염산을 사용하여 벌꿀의 pH를 2로 조절한 후 고체상 추출(SPE) 고체상인 양이온교환 카트리지에 적재한 후 염기성 메탄올로 용리하였다. 용리된 추출물은 이온쌍 시약을 사용한 이온쌍 크로마토그래피법으로 분리한 후 LC/(+)ESI-MS/MS의 MRM 방법으로 분석하였다. 정량분석을 위해서 spike 한 $5.0{\sim}250{\mu}g/kg$ 농도 범위에서 검정곡선은 좋은 직선성 ($r^2$ > 0.9951)을 나타내었다. 분석방법의 상대표준편차는 11.5~18.7%이었고 정확도는 bias로 10.9~20.9%이었다. 확립된 분석방법은 벌꿀 중에서 네오마이신의 분석방법으로 유용하게 사용될 수 있을 것이다.

• 분석과학
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• 제20권2호
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• pp.115-123
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• 2007

헤테로싸이클릭 아민류(heterocyclic amines, HAs)는 소고기, 돼지고기, 닭고기 그리고 생선과 같은 근육질의 육류를 조리 할 때 생겨나는 돌연변이원성/발암성 화학물질로 잘 알려져 있다. 식품으로부터 HAs의 분석에 있어 가장 큰 어려움은 HAs가 식품 중에 극히 미량(수 ng/g)으로 존재한다는 것과 많은 방해물질이 존재한다는 것이다. 식품으로부터 HAs를 추출하고 정제하기 위해 고체-상 추출(solid-phase extraction, SPE)이 많이 사용되고 있다. HAs 분석을 위해 여러 단계의 SPE 과정을 수행하였다. HAs의 회수율은 표준물질이 녹여져 있는 메탄올 수용액과 조리되지 않은 돼지고기에 표준용액을 스파이크하여 서로를 비교하여 얻었다. 회수율은 25.0 ng/g에서 25.3~93.0%의 값을 얻을 수 있었다. 확립된 감도 좋고 재현성있는 시료전처리방법을 통해서 ${\mu}$-LC/ESI-MS에 주입함으로써 조리된 육류로부터 HAs를 분석하는데 응용될 수 있을 것이다. 다른 방법으로는 freezing filtration 방법으로써 보다 좋은 추출 효율과 재현성을 나타내었다.

• 한국전기전자재료학회논문지
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• 제37권1호
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• pp.79-87
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• 2024

This paper is a study on the optimal microdroplet generation conditions in indirect charging electrostatic spraying. Unlike the direct charging method, which applies power to the nozzle, the indirect charging method applies power to the discharge electrode between the nozzle and the collection electrode. Therefore, an electrically simplified system can be obtained by minimizing the insulation part a stable spray pattern can be obtained with a wide spray angle, and a stable spray pattern can be obtained with a wide spray angle. To conduct the study, an indirect charging type electrostatic spray visualization system was constructed and the static characteristics of the microdroplets were analyzed through image processing of the spray shape of the microdroplets. The total number of microdroplets and the number of microdroplets per power consumption are confirmed according to the changes in the distance between the discharge electrode and the collection electrode, the flow rate, and the applied voltage, which affect the generation of microdroplets, and using this, the optimal generation conditions are derived and the corresponding microdroplet size distribution was analyzed. As a result of the experiment, it was confirmed that the optimal generation condition was at a flow rate of 15 to 20 mL/min and a voltage of -22.5 to -25 kV in terms of the number of microdroplets, and at a flow rate of 15 to 20 mL/min and a voltage of -20 kV in terms of energy consumption efficiency.

• 한국식품과학회지
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• 제41권3호
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• pp.326-333
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• 2009

국내에서 주로 섭취되는 어패류(고등어, 삼치, 조기, 대합, 바지락, 꼬막, 백합)를 가열조리 하였을 때 형성되는 HCAs를 분리, 동정하여 그 함량을 모니터링 하였다. 추출 및 정제는 고체상 추출 방법을 사용하였고 정성, 정량을 위한 분석장비는 LC/MS를 사용하였다. 모니터링 결과 생선류에서는 고등어(2.2-1,1117.7 ng/g), 삼치(9.1-443.6 ng/g), 조기(17.5-179.5 ng/g)순으로 HCAs가 많이 검출되었으며 특히 고등어 muscle에서 많은 양(1,1117.7 ng/g, Harman)이 검출되었다. 조개류 중에서는 대합(12.2-196.4 ng/g), 꼬막(2.2-76.2 ng/g), 바지락(1.7-25.8 ng/g), 백합(1.5-2.7 ng/g)순으로 많이 검출되었으며 대합의 Norharman(196.4 ng/g)이 가장 많이 검출되었다. 15종의 HCAs 중 ${beta}$-carbolines에 속하는 Norharman과 Harman이주로 검출되었으며 그 밖에 PhIP, Trp-P-1. Trp-P-2 등의 발암가능성이 있는 물질들이 검출되었다. 조리방법 측면에서 보면 간장을 이용한 조림방법보다는 직접적인 가열을 통한 구이방법이 많은 양의 HCAs를 형성시켰으며 구이 중에서도 muscle만 있는 부위에서의 검출함량보다는 muscle과 skin이 함께 있는 부위에서 검출함량이 더 많았다. 가열 조리한 어패류 분석방법에 대한 유효성을 확인해 본 결과 정량한계는 0.8-23.9 ng/mL, 검출한계는 0.2-7.2 ng/mL이었으며 회수율은 15.7-74.7%이었다.

• 생명과학회지
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• 제22권11호
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• pp.1456-1462
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• 2012

소라페닙은 멀티카이네즈 억제제로서 신세포암, 전이성 간세포암 환자의 치료에 효과가 입증된 경구용 항암제이다. 이 연구의 목적은 고속액체크로마토그래피 텐덤질량분석기법(LC/MS/MS)을 이용하여 사람 혈장 내 소라페닙의 농도를 측정하는 효율적인 방법을 개발하고 한국식품의약품안전청(KFDA) 기준에 따라 분석법을 검정하는 것이다. 혈장시료($100{\mu}l$)에 내부표준물질인 chlorantraniliprole을 첨가한 후 이소프로필알콜과 에틸아세테이트로 구성(1:4, v/v)된 0.1% 포름산 함유 추출용액을 혼합하였다. 원심분리 후 상층액을 취하여 원심감압농축하였다. 잔사를 이동상에 재용해하고 Waters사의 역상 XTerra$^{TM}$ C18 칼럼(입자크기 $3.5{\mu}m$)을 장착한 고속액체크로마토그래피 장치에 주입하였다. 액체크로마토그래피는 0.1% 포름산과 10 mM 암모늄 포메이트를 함유한 버퍼용액과 메탄올, 아세토나이트릴을 각각 1:6:3으로 혼합한 용액을 이동상으로 사용하였으며 5분 내에 측정을 완료하였다. 분석대상 물질들은 텐덤질량분석기에서 electrospray 양이온 이온화($ES^+$) 검출방식으로 확인하였으며 소라페닙은 'm/z 465.2 ${\rightarrow}$ 252.5', chlorantraniliprole은 'm/z 484.4 ${\rightarrow}$ 286.2'으로 구성한 multiple reaction monitoring 방법을 사용하였다. 검정 결과, 2-5,000 ng/ml의 농도 구간에서 양호한 직선성($r^2$ > 0.99)과 정확도(90.7-103.9%), 정밀도(10% 이하)를 나타내었다. 새롭게 개발된 LC/MS/MS을 이용한 사람 혈장 내 소라페닙의 농도 측정법은 KFDA 기준을 만족하였으며, 기존의 방법에 비해 민감도가 높은 방법이었다.

• Food Science and Biotechnology
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• 제17권3호
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• pp.475-482
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• 2008

Methanol extracts of peanut (Arachis hypogaea L.) pods were chromatographed, which yielded 3 phytochemicals 1-3 including 5,7-dihydroxychromone (1), eriodictyol (2), and 3',4',5,7-tetrahydroxyflavone (3). To confirm the presence of isolated phytochemicals, the pods extracts were performed by high performance liquid chromatography coupled with a photodiode array detector (HPLC-PDA) and a mass spectrometric detector (MSD) with electrospray ionization (ESI). Optimum extraction conditions for phytochemical contents using peanut germplasm were obtained by employing 90% MeOH for 12 hr at room temperature and phytochemicals 1-3 showed significant differences with concentrations of $407.56{\pm}23.35$, $52.92{\pm}5.11$, and $2,024.34{\pm}134.18\;{\mu}g/g$, respectively. Under this optimal conditions, the contents of phytochemicals 1-3 in peanut pods of 3 Korea cultivars including 'Jakwang', 'Daekwang', and 'Palkwang' exhibited phytochemical 3 was the highest range of $1,338.01-5,162.93\;{\mu}g/g$, followed by phytochemical 1 ($590.13-1,382.10\;{\mu}g/g$), and phytochemical 2 ($25.12-186.85\;{\mu}g/g$), respectively. Moreover, 'Jakwang' exhibited the highest contents of phytochemical (1: $1,362.10{\pm}52.49$, 2: $186.85{\pm}17.69$, and 3: $5,162.93{\pm}148.64\;{\mu}g/g$, respectively), whereas the lowest contents was found in the 'Daekwang' (1: $590.13{\pm}22.23$, 2: $25.12{\pm}2.45$, and 3: $1,338.01{\pm}62.17\;{\mu}g/g$, respectively). These results suggest that the methanol extracts of peanut pods may possess health related benefits to humans owing to various known biological activities of phytochemicals 1-3.