Evaluation of antioxidant potential of ethyl acetate fraction of Rosmarinus officinalis L. and its major components

현호봉,Sabina Shrestha,부경환,김소미 한국응용생명화학회 2015 Applied Biological Chemistry (Appl Biol Chem) Vol.58 No.5

The solvent fractions of rosemary methanol extract were obtained by successive extraction with nhexane, chloroform, ethyl acetate, butanol, and water. The ethyl acetate fraction (EAF) contained a remarkable amount of polyphenol and flavonoid as well as high levels of alkyl and 2,20-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging activity. The activity guided fractionation and repeated chromatographic separations over silica gel, RP C18, and Sephadex LH-20 led to isolation of six compounds from the EAF. 1H NMR, 13C NMR, 2D NMR, MS, and IR spectroscopies determined the compounds to be caffeic acid (1), rosmarinic acid (2), rosmarinic acid methyl ester (3), luteolin (4), apigenin (5), and hispidulin (6), and high-performance liquid chromatography quantification was used to determine concentrations in EAF. Among the six isolated compounds, rosmarinic acid methyl ester showed the highest scavenging activities against di(phenyl)-(2,4,6- trinitrophenyl) iminoazanium, alkyl and ABTS radicals. The EAF mixture, but not individual isolated compounds, shielded dermal fibroblast cells from H2O2-induced cytotoxicity at concentrations that encompass the SC50 of alkyl and ABTS radical. Therefore, our findings suggested for the first time that antioxidant capacity of the EAF mixture result in a synergistic effect on the antioxidant action.

아크릴 접착체(디클로로메탄)을 이용한 사망: 증례 보고

현호봉,박조연,박형신,홍정현,고명권,강현욱,임형수 대한법의학회 2022 대한법의학회지 Vol.46 No.4

Deaths due to dichloromethane (DCM) poisoning are mainly caused due to accidents at industrial sites, and suicidal cases are known to be rare. Herein, we report a case of a 56-year-old man who worked as a technician and died after consuming DCM for suicidal motive. According to the investigations, he had left a recorded message on his phone for his wife, which suggested he committed suicide. At the scene, opened DCM and whiskey bottles were found; approximately 120 mL of the DCM was still in the bottle. No specific injuries were externally observed during the autopsy. Microscopic examination showed denudation of epithelium in the esophageal and gastric mucosa, and no inflammatory reaction proceeded. The denudation was accompanied by pulmonary edema, acute tubular necrosis in the kidneys, and microvesicular steatosis in the liver. The DCM was detected in blood, gastric contents, and nasal cavity fluid. The concentration of alcohol in the blood was found to be 0.487%. It was presumed that he died of acute DCM poisoning while drunk. In cases of suspicious acute and chronic DCM poisoning deaths, investigations of carboxyhemoglobin and the clinical chemistry of blood or body fluids are warranted to determine the cause and mechanism of death.

Gas Chromatography-mass Spectrometry와 Headspace-Gas Chromatography-mass Spectrometry를 이용한 제주산 민트 에센셜오일 성분 분석

현호봉 ( Ho Bong Hyun ),부경환 ( Kyung Hwan Boo ),강혜림 ( Hye Rim Kang ),김소미,( Somi Kim Cho ) 한국응용생명화학회 2015 Journal of Applied Biological Chemistry (J. Appl. Vol.58 No.2

Compositions of essential oils extracted from mint herb such as Mentha piperita, Mentha spicata, and Mentha × piperita var. citrate produced in Jeju were analyzed using gas chromatography-mass spectrometry (GC-MS) and headspace- GC-MS (HS-GC-MS). By the GC-MS analysis, 13 compounds were tentatively identified in Mentha piperita, Mentha spicata, and Mentha × piperita var. citrate, respectively. Peperitenone oxide, carvone, and linalool were detected as major compounds in Mentha piperita, in Mentha spicata, in Mentha × piperita var. citrate, respectively, based on the ratio of peak intensity in the total ion chromatogram. The greater number of compounds, including volatile alcohols and acetates were identified by HSGC- MS than by GC-MS in these all three essential oils. Similarpatterns of composition were detected in both Mentha spicata and Mentha × piperita var. citrate by either one of GC-MS methods. However, in case of Mentha piperita, L-(-)-menthol, which was identified as the major compound by HS-GC-MS was detected in dramatically reduced quantity by GC-MS. Interestingly, we found that both linalyl acetate and linalool were identified as the dominant compounds in the essential oil of Mentha × piperita var. citrate.

제주 해안 노출 시기에 따른 해조류 유래 미생물 군집 구조 및 다양성 비교 분석

현호봉(Ho-Bong Hyun),김성천(Sung-Chun Kim),함영민(Young-Min Ham),정용환(Yong-Hwan Jung),권미예(Mi-Ye Kwon) 제주학회 2023 濟州島硏究 Vol.59 No.-

본 연구는 해안가로 밀려와 퇴적된 해조류의 미생물 군집 구조 및 다양성을 조사하고자 수행되었다. 본 연구는 해조류가 가장 많이 발생하는 기간 동안 제주도 서귀포시 해안의 해조류 샘플로 수행되었으며, 해수로부터 거리에 따라 해조류를 채집하였다. 채집된 해조류 샘플로부터 추출된 유전체는 차세대염기서열분석을 이용하여 메타지놈 분석을 실시하였다. 조사결과 모든 해조류 샘플에서 전체적으로 Proteobacteria와 Bacteroidetes가 가장 우점한 문(phylum)으로 확인되었다. 미생물 다양성의 경우, 구멍갈파래에서 해수에서 거리가 멀어질수록 다양성이 높아지는 것이 확인되었다. 미생물 군집구조 결과, 해안가에 밀려온 시기에 따라 다른 양상이 보였으며, 해안으로 올라 온 일정 시기의 해조류에서 alginate 분해 활성능이 있는 것으로 알려진 박테리아가 약 80% 정도의 높은 비율을 차지하고 있었다. 또한, 환경모사실험 결과 한천 분해균이 배양 7일 후에 군집 내 높은 비율을 차지하였다가 배양 18일 후에는 군집 내 비율이 줄어든 것을 확인하였다. 이러한 결과를 통해 해조류 유래 미생물 군집 구조 및 다양성 변화에 있어서 초기 군집 구조나 다양성보다 해안가에 밀려온 시기, 온도 등 환경인자가 중요한 요소가 될 수 있음을 확인할 수 있었다. 추후 생리활성 및 물리화학적 자료를 접목하고, 환경 요소에 따른 배양을 추가로 수행하여 해조류 및 이와 관련된 박테리아의 상호 기능적인 역할을 이해하고자 한다. Seaweeds have become a global industrial source material in food and cosmetic area. In recent reports, seaweed-associated bacteria play a vital role in nutrient shifts with secondary metabolites in direct and/or indirect ways. Here, we investigated the shift of bacterial communities in two different seaweed samples along with incubation and exposure duration to seashore using 16S rRNA gene-based amplicon sequencing. Bacterial communities belonging to the phyla Proteobacteria and Bacteroidetes are the most abundant on seaweeds surface. Bacterial diversity in Ulva pertusa is increased along with distances from seashore while that of Sargassum horneri are similar. Remarkably, Cobetia sp. having alginate-degrading activity represent with high proportion about 80% in. Also, the abundance of Agar-Degrading Bacteria(ADB) are fluctuated with incubation periods in mesocosm experiment, ADB predominantly present after 7 days incubation while ADB are reduced their portion after 18 days. These features would be consequences of seaweed importance to proper periods reached land. To understand the functional roles of associated bacteria in seaweeds, combined further studies should be performed coping with biochemical, physiological and microbiological approach.

Anti-melanogenesis Effects of Schizophragma hydrangeoides Leaf Ethanol Extracts via Downregulation of Tyrosinase Activity

현호봉(Ho Bong Hyun),현혜진(Hye Jin Hyeon),김성천(Sung Chun Kim),고보람(Boram Go),윤선아(Seon-A Yoon),정용환(Yong-Hwan Jung),함영민(Young-Min Ham) 한국자원식물학회 2021 한국자원식물학회지 Vol.34 No.6

Whitening agents derived from natural sources which do not have side effects are sought after. Schizophragma hydrangeoides is an edible plant that grows wild on Jeju Island. We aimed to determine whether S. hydrangeoides extracts show anti-melanogenic activity. Here, we found that 70% ethanol extracts of S. hydrangeoides leaf suppressed α-melanocytestimulating hormone-induced melanogenesis in B16F10 mouse melanoma cells. This activity of anti-melanogenesis in B16F10 cells were investigated by determining melanin content and tyrosinase activity, and by performing western blotting. The 70% ethanol extract downregulated tyrosinase and tyrosinase-related protein 1. In addition, the n-hexane fraction of S. hydrangeoides leaf (HFSH) exhibited significant anti-melanogenic activity among the various solvent fractions tested without reducing the viability of B16F10 cells. Taken together, these results indicate that extracts from S. hydrangeoides leaf can influence cellular processes via modulation of tyrosinase activity. Hence, S. hydrangeoides can be used as a whitening agent in the cosmetic industry and as a therapeutic agent for treating hyperpigmentation disorders in the clinic.

생물반응기를 이용한 황근 부정근의 대량증식과 추출물의 항산화 및 미백 활성 평가

이종두,현호봉,현혜진,장은비,고민희,윤원종,함영민,정용환,최훤,오유진,오대주 한국자원식물학회 2022 한국자원식물학회지 Vol.35 No.4

Hibiscus hamabo Sieb. et Zucc. (yellow hibiscus) is a deciduous semi-shrub plant and mainly growing in Jeju Island. This is known the unique wild hibiscus genus and classified as an 2nd grade of endangered plant for Korean Red List. In previous studies, properties of germination, ecological, genetical and salt resistance have been reported. In this study, we investigated mass-proliferated adventitious root using bioreactor, antioxidant and whitening effects to conduct functional ingredients. Yellow hibiscus were collected from Gujwa, Jeju by prior permission and they were introduced by explant type and various medium composition after surface sterilization. As a result, seed response rates were evaluated at range of 51.17 ~51.83%, in terms of comprehensive efficiency of shoot and root formation. In the case of adventitious root propagation condition was confirmed in half strength Murashige and Skoog medium salts, 30 ㎎/L sucrose, and 2 ㎎/L indole-3-butyric acid for 8 weeks in 5,000 mL bioreactor. We also compared between relationship with biomass and secondary metabolites accumulation by total phenolics content, the flavonoid content, DPPH free radical scavenging activity and melanin content. The results indicated that adventitious root mass proliferation, antioxidant and whitening effect could develop value of the high-quality cosmeceutical ingredient and further metabolite studies 황근(Hibiscus hamabo Sieb. et Zucc., 노랑무궁화)은 주로제주도 전역과 전남 남부지역에 자생하는 우리나라 유일의 야생 무궁화속의 낙엽성 반관목식물이다. 최근 무분별한 자생지파괴에 따른 개체수 감소로 환경부 지정 한국적색목록 내 멸종위기야생생물 2급으로 지정되어 있다. 선행연구에서는 황근의종자발아, 생태 및 유전적 특성, 내염성 등이 보고되었다. 따라서 본 연구에서는 생물반응기를 이용하여 황근 부정근의 대량증식, 항산화 및 미백효과를 조사하여 기능성 원료 활용 평가를 실시하였다. 황근 종자 및 식물체는 사전 환경부 채취 허가를 받은 제주시 구좌읍 관내지역에서 채집하였으며, 표면살균 후 MS 등 배지 조성을 조절하여 기내 세포주로 도입하였다. 그 결과, 종자의 전반적인 반응율이 지상부 신초 발생 및 지하부 뿌리 발근 측면에서 51.17∼51.83%로 가장 효율적인 것으로 나타났다. 5,000 mL 생물반응기에서 황근 부정근의 증식에 유리한 배지조건의 경우, 1/2 MS (Murashige and Skoog, 1962), 30 g/L 자당(sucrose), 2 ㎎/L IBA (indole-3-butyric acid)에서 8주 동안 배양한 것임을 확인하였다. 또한 총페놀 화합물 함량, 총플라보노이드 함량, DPPH 자유 라디칼 소거능 및 멜라닌 함량 분석을 통해 바이오매스 및 2차대사산물 축적의 연관관계를 비교하였다. 본 연구를 통해 황근 부정근의 대량증식, 항산화 및 미백 효능은 고부가가치 향장품 산업의 고품질 소재개발과 연계하여 추가적인 생리활성물질 연구의 가능성을 시사한다.

풋감 주정 추출물의 유전독성 연구

함영민,윤선아,현호봉,고보람,정용환,오대주,윤원종 한국식품위생안전성학회 2020 한국식품위생안전성학회지 Vol.35 No.6

감은 중국, 한국, 일본, 브라질, 터키, 이탈리아 등을 포함하는 온대지역에서 널리 재배되고 있으며 일부 아시아권 소비자들에게는 건강에 유익한 기능성 원료로 인식되고 있다. 또한 감에 포함된 풍부한 파이토케미컬들은 감을 섭취함으로써 건강과 관련된 다양한 문제점을 개선하기 위한 연구의 가능성을 제시한다. 본 연구에서는 감의 미숙과인 풋감추출물(DKA)의 유전독성을 확인하고자 한다. 미생물복귀돌연변이시험, 염색체이상시험, 포유류 소핵발생 시험을 수행하여 풋감추출물(DKA)의 유전독성을 평가하였다. 미생물복귀돌연변이시험에서 DKA는 Salmonella typhimurium TA98, TA100, TA1535, TA1537 와 Escherichia coli WP2uvrA에서 S9 대사활성계의 존재에 상관없이 돌연변이 유도를 보이지 않았다. 또한 마우스를 이용한 소핵 시험은 풋감추출물(DKA)처리군에서 소핵을 가진 다염성 적혈구와 전체적혈구 중 다염성 적혈구의 비율의 증가는 볼 수 없었으며 통계학적 유의성도 나타나지 않았다. 한편, CHL 세포를 이용한 염색체이상시험에서 모든 세포주 의 처리시간 및 S9 대사활성계 존재유무에 상관없이 염색 체이상을 보이지 않았다. 따라서 본 연구결과에 의하면 풋 감추출물(DKA)은 유전독성을 유발하지 않는 안전한 기능성 식품 원료로서 활용 가능하다고 판단된다. The persimmon is commonly cultivated in temperate regions of the world, including China, Korea, Japan, Brazil, Turkey, and Italy. In some Asian cultures, consumers are aware of the health claims related to the persimmon and its functional ingredients. The rich phytochemistry of the persimmon has opened new avenues of research on diet-based regimens designed to cure various ailments. This study was conducted to identify the genotoxicity of immature green persimmon (Diospyros kaki THUNB.) extract (DKA). The bacterial reverse mutation assay, the chromosomal aberration assay, and the mammalian micronucleus test were performed to determine the DKA genotoxicity. The result of the bacterial reverse mutation assay revealed that the DKA did not induce mutagenicity in Salmonella typhimurium TA98, TA100, TA1535, TA1537 and Escherichia coli WP2uvrA with or without metabolic activation of S9 mixture. The oral administration of DKA also caused no significant increase in the number of micronucleated polychromatic erythrocytes or in the mean ratio of polychromatic to total erythrocytes. In addition, DKA did not cause a significant chromosome aberration on CHL cells in the presence or absence of S9 activation. In conclusion, DKA could be considered as a reliable and safe functional food since no toxicity was found under the condition of this study.

붉은인동(Lonicera periclymenum) 배발생세포 유래 캘러스, 부정근, 식물체 배양시스템에 따른 바이오매스 및 항산화 기능성 평가

이종두(Jong-Du Lee),현호봉(Ho Bong Hyun),장은비(Eunbi Jang),윤원종(Weon-Jong Yoon),함영민(Young Min Ham),정용환(Yong-Hwan Jung),박소영(So-Young Park),권아름(A-Reum Kwon) 한국원예학회 2020 한국원예학회 학술발표요지 Vol.2020 No.11

단메밀과 쓴메밀의 ITS 염기서열 기반 식별마커

오대주 ( Oh Dae-ju ),현호봉 ( Hyun Ho-bong ),임태준 ( Lim Tae-joon ),윤선아 ( Yoon Seon-a ),함영민 ( Ham Young-min ),윤원종 ( Yoon Weon-jong ),양우삼 ( Yang Woo-sam ),정용환 ( Jung Yong-hwan ) 한국유기농업학회 2018 韓國有機農業學會誌 Vol.26 No.4

We analyzed the nuclear ribosomal internal transcribed spacer (ITS) sequence of common buckwheat, Fagopyrum esculentum and tartary buckwheat, F. tataricum. The diversity of the nucleotides and haplotypes, Tajima’s D, and Fu’s Fs was analyzed and compared among the varieties of common buckwheat and tartary buckwheat. The diversity of nucleotides and haplotypes indicated that the buckwheat populations had undergone rapid population expansion but D and Fs did not support their expansion statistically. The phylogenetic analysis of ITS sequences did not clearly establish the phylogenetic relationships between the varieties of common buckwheat. The In/Del sequence of ITS-1 region could, therefore, be used as a DNA marker to distinguish raw or manufactured products derived from common buckwheat and tartary buckwheat.

인간 유방암 줄기세포에서 레몬잎 메탄올 추출물의 항암 효능

문정용,( Linh Thi Thao Nguyen ),현호봉,( Ahmed Osman ),조민환,한수영,이동선,안광석 한국응용생명화학회 2015 Journal of Applied Biological Chemistry (J. Appl. Vol.58 No.3

The anticancer activity of a methanolic extract from lemon leaves (MLL) was assessed in MCF-7-SC human breast cancer stem cells. MLL induced apoptosis in MCF-7-SC, as evidenced by increased apoptotic body formation, sub-G1 cell population, annexin V-positive cells, Bax/Bcl-2 ratio, as well as proteolytic activation of caspase-9 and caspase-3, and degradation of poly (ADP-ribose) polymerase (PARP) protein. Concomitantly, MLL induced the formation of acidic vesicular organelles, increased LC3-II accumulation, and reduced the activation of Akt, mTOR, and p70S6K, suggesting that MLL initiates an autophagic progression in MCF-7-SC via the Akt/mTOR pathway. Epithelialmesenchymal transition (EMT), a critical step in the acquisition of the metastatic state, is an attractive target for therapeutic interventions directed against tumor metastasis. At low concentrations, MLL induced anti-metastatic effects on MCF-7-SC by inhibiting the EMT process. Exposure to MLL also led to an increase in the epithelial marker E-cadherin, but decreased protein levels of the mesenchymal markers Snail and Slug. Collectively, this study provides evidence that lemon leaves possess cytotoxicity and antimetastatic properties. Therefore, MLL may prove to be beneficial as a medicinal plant for alternative novel anticancer drugs and nutraceutical products.