머루 줄기와 자소자로부터 분리한 Resveratrol 올리고머와 Flavonoid의 항균효과

손락호(Rak Ho Son),진휘승(Hwi Seung Chin),함아름(Ahrom Ham),마응천(Woongchon Mar),남궁우(Kung-Woo Nam) 대한약학회 2010 약학회지 Vol.54 No.1

We studied the antimicrobial activities of five compounds isolated from the stems of Vitis coignetiae Pulliat and the seeds of Perilla frutescens (L.) Britton. Based on spectroscopic evidence, compounds 1 to 5 were characterized as resveratrol, ε-viniferin, ampelopsin E, apigenin, and luteolin, respectively. The antimicrobial activities against Gram-positive (Staphylococcus aureus) and -negative (Pseudomonas aeruginosa) bacteria and a fungus (Candida albicans) were investigated using the disc diffusion and broth dilution methods. C. albicans was not inhibited by the five compounds. Compounds 2 and 5 had significant anti-microbial activity against S. aureus, and the 50% inhibitory concentration (IC50) of compound 2 against S. aureus was 7.2 μM. Compounds 4 and 5 significantly inhibited P. aeruginosa and the minimum inhibitory concentration (MIC) of compounds 2 and 5 was 0.07 and 2.0 μM, respectively. Compounds 2, 4, and 5 had strong anti-microbial activity against S. aureus and P. aeruginosa.

Pectinase를 이용한 복분자, 오디, 아로니아 과즙의 항산화 활성

손락호(Son Rak Ho),채규서(Chae Kyu Seo),김기안(Kim Ki An),권지웅(Kwon Ji Wung),박재숙(Park Jae Suk),조하니(Cho Ha Ni) 한국약용작물학회 2014 한국약용작물학회 학술대회논문집 Vol.2014 No.2

토종효모를 이용한 복분자 발효주의 바이오제닉아민 함량 및 향기성분 분석

윤해훈,손락호,류은혜,정지혜,Yoon, Hae-Hoon,Son, Rak-Ho,Ryu, Eun-Hye,Jung, Ji-Hye 한국식품조리과학회 2015 한국식품조리과학회지 Vol.31 No.3

We investigate black raspberry (Rubus occidentalis) wine made using traditional yeast (Saccharomyces cerevisiae A8, B6, GBY2, GBY3) and S. cerevisiae Fermivin (FM), which is widely used in wine manufacturing, and analyze the biogenic amine content and the volatile flavor compounds. Black raspberries were separately inoculated with yeast up to $1{\times}10^9CFU/kg$, followed by incubation at $25^{\circ}C$ for 7 days. FM produced the highest alcohol content, however the final fermentation characteristics of the wine made using four different yeasts were similar. S. cerevisiae A8 had a large biogenic amine (BA) content, specifically tryptamine, thus we excluded this yeast from fermentation. S. cerevisiae GBY3 was selected for black raspberry wine fermentation as a result of sensory evaluation. The volatile flavor compounds of two wines (S. cerevisiae GBY3 and FM) were analyzed by gas chromatography and mass spectrometry. 37 compounds in the samples were separated, and several ester compounds were identified in greater amounts in the wine made with S. cerevisiae GBY3 than in the wine made with FM. A greater amount of the major compound, ethyl benzoate, giving the sweet and fruity flavor, was identified in wine made with S. cerevisiae GBY3 than in the wine made with FM. In conclusion, S. cerevisiae GBY3 was confirmed to produce no major BAs and a better flavored wine. These results give new leads in the production of high quality wine.

원지와 영여자로부터 분리한 8종 화합물의 항균효과

진휘승(Hwi Seung Chin),손락호(Rak Ho Son),이용화(Yong-Hwa Lee),함아름(Ahrom Ham),마응천(Woongchon Mar),김원기(Won-Ki Kim),남궁우(Kung-Woo Nam) 대한약학회 2010 약학회지 Vol.54 No.2

The chemical structures of eight compounds purified from two plants (Polygala tenuifolia Willdenow and Dioscorea batatas Decene) were determined and their anti-microbial activity against three microbial strains (Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans) was tested. The three micro organisms were cultured in 96-well plates or Petri dishes without (control) or with the eight compounds added at concentrations of 100 to 0.01 μM (wt/vol). The growth of the microorganisms in the medium was examined after a 24-h incubation. The inhibitory effect of each compound on the growth of the microorganisms was calculated from the optical density measured at 595 nm, turbidity, and size of the inhibition zone around the treated paper disc. The minimum inhiitory concentration (MIC) of compounds 4 to 7 against S. aureus was 0.08, 0.05, 1.3 and 0.02 μM, respectively, and 0.09, 0.1, 0.2 and 100 μM against C. albicans. The IC50 (50% inhibition) values of compounds 5 and 6 were 3.1 and 6.4 μM against S. aureus, respectively, and 10 and 2.4 μM against C. albicans. Therefore, compounds 4 to 6 were the most potent anti-microbial agents among the eight compounds tested.

복령피 추출물의 멜라닌 생성 저해 효과

이응지 ( Eung-ji Lee ),배성윤 ( Seong Yun Bae ),손락호 ( Rak Ho Son ),이용화 ( Yong Hwa Lee ) 대한화장품학회 2009 대한화장품학회지 Vol.35 No.3

천연 미백 소재 개발을 위하여 복령피 추출물과 복령피 추출물에서 분리한 활성 물질의 멜라닌 생성에 연관된 생리 활성을 분석하였다. 복령피 추출물은 100 μg/mL 이하에서 세포 독성이 없는 것으로 확인 되었으며 free radical 소거능(DPPH)과 superoxide radicals 소거능 결과는 각각 IC₅₀ = 19.4 ± 2.21 μg/mL, IC₅₀ = 103 ± 3.33 μg/mL을 나타내었다. 50 μg/mL 농도에서 B16 melanoma 세포 내 tyrosinase의 활성을 34 % 저해 하였으며, 세포 내 멜라닌 생합성 저해 효과는 50 μg/mL 농도의 복령피 추출물을 72 h 동안 처리한 세포에서 51 % 억제율을 보이는 것으로 나타났다. 이러한 복령피 추출물의 활성 물질을 분리하여 1H-NMR, 13C-NMR, Mass analysis 등의 기기 분석을 실시한 결과 triterpene류의 3-β-hydroxylanosta-7,9(11),24-trien-4-oic acid로 동정되었고 100 μg/mL 이하에서 세포 독성이 없는 것으로 확인 되었다. 3-β-hydroxylanosta-7,9(11),24-trien-4-oic acid의 free radical 소거능과 superoxide radicals 소거능 결과는 각각 IC₅₀ = 4.3 ± 0.15 μg/mL, IC₅₀ = 54 ± 1.67 μg/mL을 나타내었다. 10 μg/mL 농도에서 세포내 tyrosinase의 활성을 43 % 저해 하였으며, 멜라닌 저해 효과를 확인한 결과 IC₅₀ = 3.6 μg/mL으로 나타났다. Western blot을 이용하여 tyrosinase, tyrosinase related protein-1 (TRP-1), TRP-2 단백질의 발현 감소를 확인하였다. 복령피 추출물과 3-β-hydroxylanosta-7,9(11),24-trien-4-oic acid는 우수한 미백 효능을 갖는 화장품 소재로의 개발 가능성이 클 것으로 기대된다. To develop a new natural whitening agent for cosmetics, we investigated the inhibitory effects of Poria cocos Bark extracts (PCBE) and its active compound on melanogenesis. PCBE showed ROS scavenging activities in 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and xanthine/xanthine oxidase system with the IC₅₀ values of 19.4 ± 2.21 μg/mL and IC₅₀ = 103 ± 3.33 μg/mL, respectively. PCBE reduced intracellular tyrosinase activity about 34 % at concentration of 50 μg/mL. And PCBE reduced melanin contents of B16 melanoma cells about 51 % at concentration of 50 μg/mL without cell cytotoxicity (below 100 μg/mL). We purified one active compound from PCBE and identified its structure. It was identified as 3-β-hydroxylanosta-7,9(11),24-trien-4-oic acid, triterpene family, by 1H-NMR, 13C-NMR and Mass analysis. 3-β-hydroxylanosta-7,9(11),24-trien-4-oic acid showed ROS scavenging activities in DPPH radical and xanthine/xanthine oxidase system with the IC₅₀ values of 4.3 ± 0.15 μg/mL and 54 ± 1.67 μg/mL, respectively. Also, it was shown that 3-β-hydroxylanosta-7,9(11),24-trien-4-oic acid reduced intracellular tyrosinase activity about 43 % at concentration of 10 μg/mL. And it inhibited melanin synthesis in a dose dependent manner (IC₅₀ = 3.6 μg/mL) without cell cytotoxicity (below 100 μg/mL). 3-β-hydroxylanosta- 7,9(11),24-trien-4-oic acid inhibited tyrosinase, TRP-1 and TRP-2 expression at protein level. These results suggest that PCBE and 3-β-hydroxylanosta-7,9(11),24-trien-4-oic acid reduced melanin formation by the inhibition of tyrosinase activity and expression in B16 melanoma cells. Therefore, we suggest that PCBE could be used as a useful whitening agent.

사군자추출분말(HU033)의 Quisqualic Acid 성분 분석법 검증

김명일(Myeong-Il Kim),윤지수(Ji-Soo Yoon),오명환(Myeong-Hwan Oh),손락호(Rak-Ho Son),연성흠(Sung-Hum Yeon) 한국식품영양과학회 2021 한국식품영양과학회지 Vol.50 No.2

사군자추출분말(HU033)을 식약처 건강기능식품 개별인정 원료로 개발하기 위한 연구의 일환으로 원료 표준화를 위해 지표성분인 quisqualic acid의 HPLC 분석법을 설정하였고 이에 대한 검증을 실시하였다. 그 결과 표준용액과 HU033의 HPLC 크로마토그램을 비교하여 다른 물질의 간섭 없이 피크가 선택적으로 분리되었으며, 표준용액과 HU033의 피크 머무름시간이 정확히 일치하는 것을 확인하였다. 또한 동일한 스펙트럼을 나타내는 것을 확인하여 본 시험법의 특이성을 확인하였다. 검량선의 상관계수(R2)는 모두 0.998 이상으로 높은 직선성을 확인하였고, 직선성의 검출한계는 0.976 ㎍/mL, 정량한계는 2.958 ㎍/mL로 확인되었다. HU033에 1.25~10 ㎍/mL 농도로 표준물질을 첨가하여 조제한 용액을 동일한 조건으로 반복 실험하여 시험한 결과, 전체 평균회수율은 100.26%로 분석의 높은 정확성을 확인하였다. HU033의 검체량을 달리하여 각각 5회 반복 분석한 결과 전체 RSD(%) 구간은 0.52~1.90%로 확인되었고, quisqualic acid 분석의 정밀성을 확인하고자 intra-day와 inter-day 정밀성을 확인하였다. Intra-day 정밀성은 0.24~0.41%, inter-day 정밀성은 0.02~0.66% 수준으로 확인되어 높은 정밀성을 나타내었다. 따라서 본 연구를 통해 HU033의 지표성분 quisqualic acid 분석법은 적합한 시험법임을 검증하였으며 향후 기능성 원료 개발을 위한 기초자료로 활용될 것으로 사료된다. In the current study, high performance liquid chromatography was used to validate and establish a method for identification of quisqualic acid (QA) in Quisqualis indica extract powder (HU033), which is widely used in health functional foods. Sample preparation was achieved by applying o-phthalaldehyde derivatization and protein hydrolysis. Analysis was performed on a C18 column with a gradient elution containing a mixture of sodium acetate (0.01 M, pH 4.6) and acetic acid (0.2 M) in water, and a flow rate of 1.0 mL/min methanol, detected at 338 ㎚. Method validation was achieved by measuring the specificity, linearity, accuracy and precision. Calibration curve of QA showed high linearity (R²=1), and limits of detection and quantification were determined to be 1.0 and 3.0 ㎍/mL, respectively. Recovery rate ranged from 98.66% to 101.11%, and QA content in HU033 was approximately 1.09%. Based on these results, we conclude that the developed method can be applied for quantifying QA in Quisqualis indica extract powder (HU033).

토종발효미생물을 이용한 오디 발효주의 항산화 활성 및 향기성분 분석

채규서(Kyu Seo Chae),정지혜(Ji Hye Jung),윤해훈(Hae Hoon Yoon),손락호(Rak Ho Son) 한국식품영양과학회 2014 한국식품영양과학회지 Vol.43 No.7

This study was carried out to develop mulberry wines fermented with traditional microorganisms (Saccharomyces cerevisiae B-8). S. cerevisiae B-8 is a traditional fermentation microorganism isolated from domestically grown Rubus occidentalis. Each S. cerevisiae B-8 and Fermivin was inoculated into mulberry up to 1×10<SUP>9</SUP> CFU/kg, followed by incubation at 25°C for 10 days. Mulberry fermented with S. cerevisiae B-8 (MBB) had a high alcohol content (16.47%), and the fermentation rate of MBB was faster than that of mulberry fermented with Fermivin (MBF). The total polyphenol and flavonoid contents of MBB were higher than those of MBF. DPPH radical scavenging activity of MBB was as high as that of MBF. ABTS radical scavenging activity of MBF was higher than those of MBB and mulberry juice (MBJ). In addition, reducing power of MBB was much higher than other samples. Flavor constituents of the two fermented wines were analyzed by gas chromatography and mass spectrometry. Twenty-three compounds from the sample were separated and identified as fifteen esters, six alcohols, an aldehyde, and an acetate. Particularly, tetradecanoic acid, ethyl ester of orris and violet flavor were ten times more abundant in MBB than in MBF. Several ester components were two times more abundant in MBB than in MBF. In conclusion, current findings indicate that MBB might have better antioxidant activities with flavor, which contributes to improved wine production with high quality and function.

광곽향 추출물의 멜라닌 생성 저해 효과

배성윤 ( Seong Yun Bae ),이응지 ( Eung Ji Lee ),손락호 ( Rak Ho Son ),이용화 ( Yong Hwa Lee ) 대한화장품학회 2009 대한화장품학회지 Vol.35 No.1

본 연구는 천연 미백소재 개발을 위하여 광곽향 추출물과 광곽향 추출물에서 분리한 활성물질의 멜라닌 생성에 연관된 생리활성을 분석하였다. 광곽향 추출물은 100 μg/mL 이하에서 세포 독성이 없는 것으로 확인되었으며 free radical 소거능(DPPH)과 superoxide radical 소거능 결과는 각각 IC₅₀ = 24.2 ± 2.85 μg/mL, IC₅₀ = 118 ± 0.43 μg/mL을 나타내었다. B16 melanoma 세포에서의 멜라닌 생합성 저해 효과는 20 μg/mL 농도의 광곽향 추출물을 72 h 동안 처리한 세포에서 멜라닌 억제율이 23 %로 나타났으며, 50 μg/mL 농도에서 세포 내 tyrosinase의 활성을 18 % 저해하였다. 이러한 광곽향 추출물의 활성물질을 분리하여 ¹H-NMR, ¹³C-NMR, Mass analysis 등의 기기분석을 실시한 결과 sesquiterpene 계열의 활성물질인 patchouli alcohol으로 동정되었고, patchouli alcohol의 free radical 소거능과 superoxide radical 소거능 결과는 각각 IC₅₀ = 3.14 ± 0.12 μg/mL, IC₅₀ = 49 ± 3.24 μg/mL을 나타내었다. 또한 멜라닌 저해효과를 확인한 결과 IC₅₀ = 3.9 μg/mL으로 나타났으며, 10 μg/mL 농도에서 세포 내 tyrosinase의 활성을 40 % 저해하였다. Western blot을 이용하여 tyrosinase와 tyrosinase related protein-2 (TRP-2) 단백질의 발현 감소를 확인하였다. 그러므로 광곽향 추출물과 patchouli alcohol은 우수한 미백 효능을 갖는 화장품 소재로서 개발 가능성이 클 것으로 기대된다. To develop a new natural whitening agent for cosmetics, we investigated the inhibitory effects of Pogostemon cablin Bentham extracts (PCE) and its active component on melanogenesis. PCE showed ROS scavenging activities in 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and xanthine/xanthine oxidase system with the IC₅₀ values of 24.2 ± 2.85 μg/mL and IC₅₀ = 118 ± 0.43 μg/mL, respectively. PCE reduced melanin contents of B16 melanoma cells in a dose-dependant manner and decreased to about 23 % at a concentration of 20 μg/mL without cell cytotoxicity (below 100 μg/mL). And the PCE reduced intracellular tyrosinase activity about 18 % at concentration of 50 μg/mL. We purified one active compound from PCE and identified its structure. It was identified as patchouli alcohol, sesquiterpene family, by ¹H-NMR, ¹³C-NMR, and Mass analysis. Patchouli alcohol also inhibited ROS scavenging activities in DPPH radical and xanthine/xanthine oxidase system with the IC₅₀ values of 3.14 ± 0.12 μg/mL and 49 ± 3.24 μg/mL, respectively. Patchouli alcohol inhibited melanin synthesis in a dose dependent manner (IC₅₀ = 3.9 μg/mL). And the patchouli alcohol reduced intracellular tyrosinase activity about 40 % at concentration of 10 μg/mL. Patchouli alcohol inhibited tyrosinase and TRP-2 expression at protein level. These results suggest that PCE and patchouli alcohol reduced melanin formation by the inhibited of tyrosinase activity and expression in B16 melanoma cells. Therefore, we suggest that PCE could be used as a useful whitening agent.

복분자 식초의 품질특성과 항산화활성

박송이 ( Song Yie Park ),채규서 ( Kyu Seo Chae ),손락호 ( Rak Ho Son ),정지혜 ( Ji Hye Jung ),임영례 ( Young Rae Im ),권지웅 ( Ji Wung Kwon ) 한국산업식품공학회 2012 산업 식품공학 Vol.16 No.4

본 연구에서는 2단계 발효를 통해 복분자 식초를 제조한 후 이에 대한 이화학적 성분 및 항산화 활성을 측정하였다. 복분자 식초의 알코올 발효 종료 후 알코올 함량은 11.78±0.03%이었고, 당도는 8.40±0.05 Brix이었으며, 초산발효 종료 후 알코올 함량은 0.16±0.03%이었고, 산도는 6.07±0.11%이었다. 복분자 식초의 주요 유기산은 acetic acid(4,641.59 mg%)와 citric acid(1,241.05 mg%)이었고, 복분자 식초의 총 폴리페놀, 총 플라보노이드 함량은 각각 38.01 mg/g, 17.91 mg/g이었다. 페놀화합물 중 ellagic acid 함량이 1,127.43ug/g으로 가장 많았으며, gallic acid의 함량은 962.44ug/g을 보였다. 복분자 식초의 DPPH 라디칼소거 활성과 환원력은 500ug/mL 농도에서 각각 60.3%, 0.50이었다. This study was performed to investigate the physicochemical properties and antioxidant activity of bokbunja (Black raspberry) vinegar by two step fermentation. The bokbunja vinegar was contained the highest amount of acetic acid (4,641.59 mg%) followed by citric acid (1,241.05 mg%) among the major organic acids in the vinegar. The total polyphenol and flavonoid contents in bokbunja vinegar were 38.0 mg/g and 17.8 mg/g, respectively. The ellagic acid and gallic acid contents in bokbunja vinegar were 1,127.43ug/g and 962.44ug/g, respectively. At concentration of 500ug/mL bokbunja vinegar, the DPPH radical scavenging activity and reducing power were 60.3% and 0.50, respectively.

토종효모를 이용한 블루베리 발효주의 발효 특성 및 항산화 활성

윤해훈(Hae Hoon Yoon),채규서(Kyu Seo Chae),손락호(Rak Ho Son),정지혜(Ji Hye Jung) 한국식품영양과학회 2015 한국식품영양과학회지 Vol.44 No.6

본 연구에서는 국내 복분자로부터 분리된 토종효모 Saccharomyces cerevisiae M-5와 수입 시판건조효모 Fermivin을 이용한 블루베리의 발효 품질 특성을 확인하였고, 항산화 활성을 확인하기 위해 총 폴리페놀 및 총 플라보노이드, DPPH 라디칼 소거 활성, ABTS 라디칼 소거 활성, 환원력을 측정하였다. S. cerevisiae M-5와 Fermivin을 블루베리에 1×10<SUP>9</SUP> CFU/kg이 되도록 접종하여 25°C에서 39일간 발효를 진행하였다. S. cerevisiae M-5를 이용한 블루베리 발효주(BBM)의 최종 품질은 알코올 13.10%, 당도 8.42°Bx, 산도 1.183%였으며, Fermivin을 이용한 블루베리 발효주(BBF)는 알코올 14.20%, 당도 8.2°Bx, 산도 1.153%로 발효가 완료되었다. 발효 특성은 BBM이 BBF와 비교 시 초기발효 속도가 빠르며 최종 알코올 생성량은 약 1% 낮은 것으로 확인되었다. 총 폴리페놀 함량은 BBJ가 0.74±0.07 TAE mg/mL, BBM이 1.34±0.20 TAE mg/mL, BBF가 1.09±0.01 TAE mg/mL로 발효 시 증가되었으며, BBM의 총 폴리페놀 함량이 BBF보다 약 1.2배 높게 확인되었다. 총 플라보노이드 함량은 BBM이 0.61±0.20 QUE mg/mL, BBF가 0.55±0.08 QUE mg/mL로 발효하였을 때 증가하며 BBM의 총 플라보노이드 함량이 높음을 확인하였다. 항산화 활성확인을 위한 DPPH 라디칼 소거 활성, ABTS 라디칼 소거활성 그리고 환원력 또한 총 폴리페놀 및 총 플라보노이드 함량이 높았던 BBM이 더 높은 항산화 활성을 나타내었다. 효모를 달리하여 제조된 블루베리 발효주의 관능검사를 실시한 결과 색상의 경우 BBF가 높았으나 향미, 맛, 전체적인 기호도는 BBM이 우수한 기호도를 보였다. 결과적으로 토종효모 S. cerevisiae M-5를 이용한 발효주는 수입 시판건조효모 Fermivin을 이용한 발효주보다 항산화 활성 및 관능적 기호도에서 우수하게 평가되어 토종효모 S. cerevisiae M-5는 와인 제조 등과 같은 산업적 활용이 가능하고 이용가치가 높을 것으로 판단된다. We investigated blueberry wine made using traditional yeast (Saccharomyces cerevisiae M-5) and Saccharomyces cerevisiae Fermivin which is widely used in wine manufacturing, and measured its fermentative characteristics and antioxidant activity. S. cerevisiae M-5 is a traditional yeast isolated from domestically grown Black raspberry (Rubus occidentalis). Both S. cerevisiae M-5 and Fermivin were inoculated into blueberry juice (BBJ) up to 1×10<SUP>9</SUP> CFU/kg, followed by incubation at 25°C for 39 days. Final fermentation products of blueberry fermented with S. cerevisiae M-5 (BBM) presented 13.10% alcohol, 8.42°Bx of sugar, and 1.183% acidity, and final fermentation products of blueberry fermented with Fermivin (BBF) presented 14.20% alcohol, 8.2°Bx of sugar, and 1.153% acidity. The contents of total polyphenol and flavonoid compounds of BBM were higher than those of BBF and BBJ. DPPH and ABTS radical scavenging activities and reducing power of BBM were higher than those of BBF. The sensory evaluation revealed that BBM showed excellent flavor, taste, and overall acceptability compared with BBF. Based on these results, the possibility of industrial utilization of S. cerevisiae M-5 as traditional yeast was confirmed by sensory evaluation and antioxidant activity. Fermentation rate of S. cerevisiae M-5 was similar to Fermivin, which is used in the food industry.