• Journal of Applied Biological Chemistry
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• 제54권2호
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• pp.114-119
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• 2011

황백으로부터 n-BuOH 추출물에서 약 50%의 hyaluronidase 저해효과가 있음이 확인되었다. n-BuOH 분획 농축물을 Sephadex LH-20 및 MCI gel CHP-20 column chromatography로 정제한 결과 저해활성을 가지는 Jatrorrhizine을 분리, 동정하였다. 황백추출물과 Jatrorrhizine은 모두 농도 의존적으로 NO 발현을 억제하였고, 100 ${\mu}g/mL$의 농도에서 60% 이상의 억제 효과를 나타내었다. 황백추출물과 Jatrorrhizine의 iNOS와 COX-2의 억제력을 살펴본 결과, 100 ${\mu}g/mL$의 농도에서 45% 및 29%의 저해효과를 나타내었다. 따라서 황백추출물 및 Jatrorrhizine은 LPS로 유도되어진 대식세포주인 Raw264.7 세포에서의 염증반응의 억제효과를 기대할 수 있었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.40-44
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• 2003

Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

• Journal of Ginseng Research
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• 제44권3호
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• pp.405-412
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• 2020

Background: Root rot is a serious destructive disease of Panax notoginseng, a famous cultivated araliaceous herb called Sanqi or Tianqi in Southwest China. Methods: The chemical substances of Sanqi rot roots were explored by chromatographic techniques. MS, 1D/2D-NMR, and single crystal X-ray diffraction were applied to determine the structures. Murine macrophage RAW264.7 and five human cancer cell lines were used separately for evaluating the antiinflammatory and cytotoxic activities. Results and Conclusion: Thirty dammarane-type triterpenes and saponins were isolated from the rot roots of P. notoginseng. Among them, seven triterpenes, namely, 20(S)-dammar-25-ene-24(S)-hydroperoxyl-3β,6α,12β,20-tetrol (1), 20(S)-dammar-3-oxo-23-ene-25-hydroperoxyl-6α,12β,20-triol (2), 20(S)-dammar-12-oxo-23-ene-25-hydroperoxyl-3β,6α,20-triol (3), 20(S)-dammar-3-oxo-23-ene-25-hydroperoxyl-12β,20-diol (4), 20(S),24(R)-epoxy-3,4-seco-dammar-25-hydroxy-12-one-3-oic acid (5), 20(S),24(R)-epoxy-3,4-seco-dammar-25-hydroxy-12-one-3-oic acid methyl ester (6), and 6α-hydroxy-22,23,24,25,26,27-hexanordammar-3,12,20-trione (7), are new compounds. In addition, 12 known ones (12-16 and 19-25) were reported in Sanqi for the first time. The new Compound 1 showed comparable antiinflammatory activity on inhibition of NO production to the positive control, whereas the known compounds 9, 12, 13, and 16 displayed moderate cytotoxicities against five human cancer cell lines. The results will provide scientific basis for understanding the chemical constituents of Sanqi rot roots and new candidates for searching antiinflammatory and antitumor agents.

• Applied Biological Chemistry
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• 제51권2호
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• pp.142-147
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• 2008

Rhizopus oryzae KSD-815를 밀에 접종하여 만든 누룩(Rhizopus oryzae KSD-815)에서 4종의 지질 화합물을 분리하여 화학구조를 동종하였다. 건조된 누룩을 실온에서 80% MeOH 수용액으로 추출하고 이추출물을 EtOAc 분획, n-BuOH 분획, $H_2O$ 분획으로 나누었다. EtOAc분획은 다시 80% MeOH 과 n-hexane 으로 분획하였다. n-Hexane 분획에 대해 silica gel 및 ODS column chromatography를 반복 실시하여 4종의 지질 화합물을 분리, 정제하였다. NMR, IR, GC/MS 등을 통하여 화합물 1(linolenic acid methyl ester), 화합물 2(palmitic acid methyl ester), 화합물 3(linoleic acid), 화합물 4(palmitic acid)의 구조를 결정하였다. 이 지방산 화합물의 세포독성을 평가하기 위해 인체 유래 유방암(MDA-MB-231)과 간암(SK-HEP-1)세포주에 대해 MTT assay를 수행하였다. 두 암 세포주에서 화합물 1(linolenic acid methyl ester)과 화합물 3(linoleic acid)은 농도의존적인 세포독성을 확인하였다. 또 염증반응 매개체의 일종인 nitric oxice(NO)의 생성 억제 활성을 Griess 방법으로 평가한 결과, 화합물 3(linoleic acid)은 LPS와 IFN-${\gamma}$에 의해 유도 된 NO 생성도 저해하는 것을 RAW264.7 세포에서 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제8권5호
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• pp.455-460
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• 1998

In the course of screening for new antitumor substances, a novel cytotoxic agent inducing apoptotic cell death was isolated from the culture broth of marine bacterial strain SCRC A-20. Strain SCRC A-20 was separated from a mollusk and was chemotaxonomically identified as a Streptomyces sp. The cytotoxic substance was purified by organic solvent extraction followed by silica gel column chromatography and preparative TLC. HRFAB-MS determined its molecular formula to be $C_{30}H_{40}N_2O_5$ (MW 508). The 1D and 2D NMR spectral data demonstrated that the substance has a novel lactam structure of a 20-membered macrocycle coupled with a unique acyl tetramine and bicyclo［3.3.0］ octane, which includes three methyl groups, six olefinic protons, five carbonyl groups, a conjugated diene and a dienone. The substance, named aburatubolactam C, appeared to be cytotoxic for various continuously proliferating tumor cells of human and murine origins. The $IC_{50}$ values determined by MIT assay were in the range of 0.3 to $5.8\mug/ml$. When Jurkat T cells were treated with $3\mug/ml$. of aburatubolactam C, the apoptotic DNA fragmentation was detectable within 3 h, indicating that the cytotoxic effect of aburatubolactam C on tumor cells is attributable to the induced apoptosis.

• Journal of Microbiology and Biotechnology
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• 제20권6호
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• pp.988-994
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• 2010

The EtOAc fraction of Lespedeza cyrtobotrya showed mushroom tyrosinase inhibitory and radical scavenging activities. Four active compounds were isolated based on Sephadex LH-20 chromatography and HPLC, and the structures were elucidated, on the basis of their LC-MS and NMR spectral data, as 2-(2,4-dihydroxyphenyl)-6-hydroxybenzofuran (1), eriodictyol-7-O-glucopyranoside (2), haginin A (3), and dalbergioidin (4), respectively. Compound (1) showed mushroom tyrosinase inhibitory activity with an $IC_{50}$ value of $5.2\;{\mu}M$ and acted as a competitive inhibitor. Furthermore, $37.3\;{\mu}M$ of compound 1 reduced 50% of the melanin content on human melanoma (MNT-1) cells. The radical scavenging activities of compounds 1, 2, 3, and 4 were shown to have $IC_{50}$ values of 11.0, 24.5, 9.0, and $36.5\;{\mu}M$, respectively, in an ABTS system and $IC_{50}$ values of 42.7, 36.0, 37.7, and $61.7\;{\mu}M$, respectively, in a DPPH system. The mushroom tyrosinase inhibitory activity of the EtOAc fraction of Lespedeza cyrtobotrya was contributed by compounds 1, 3, and 4, and its radical scavenging activity was contributed by compounds 1-4.

• Journal of Applied Biological Chemistry
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• 제57권1호
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• pp.47-51
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• 2014

등심초(Juncus effusus)를 70% ethanol로 추출하고 이 추출물을 ethyl acetate분획, n-butanol 및 $H_2O$ 분획으로 나누었다. Ethyl acetate 분획에 대하여 silica gel, Sephadex LH-20 column chromatography를 반복 실시하여 2종의 화합물을 분리하였다. 화합물 1 (dehydroeffusol)과 2 (effusol)에 대하여 free radical 소거활성을 측정한 결과, dehydroeffusol (DPPH; $130{\pm}3.21{\mu}M$, ABTS; $39{\pm}3.51{\mu}M$)과 effusol (DPPH; $79{\pm}1.53{\mu}M$, ABTS; $24{\pm}2.73{\mu}M$)은 강한 항산화 활성을 나타내었다. 또한 이들 화합물들의 인체유래 위암세포(AGS), 폐암세포(A549)에 대한 성장억제 효과를 측정한 결과 모두 항암약물(CPA)과 유사하거나 더 높은 성장 저해 효과를 보였다.

• 대한화학회지
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• 제35권6호
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• pp.707-712
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• 1991

수은 pool 전해조를 사용하여 (5:3) EtOH/4N-HCl 용액계에서 선택적인 -N-N=O기의 전기화학적 환원에 의해 N'-phenyl-N-(2-chloroethyl)-N-nitrosourea로부터 N'-phenyl-N-(2-chloroethyl)-N-aminourea를 합성하였다. 전기화학적 환원에 앞서 반응의 최적조건을 검토하기 위하여 몇 가지 N'-aryl-N-(2-chloroethyl)-N-nitrosourea 유사체에 대하여 pH변화에 따른 환원전위를 순환전압전류법으로 조사한 결과 pH에 따라 $E_p$값이 "-"쪽으로 이동하였으며 aryl기의 치환기 효과는 -N-N=O기의 환원 전위에 크게 영향을 미치지 않는 것으로 나타났다. 용액계의 pH에 따른 반응성을 조사한 결과 강산성 용액(pH<1)에서 반응이 가장 잘 진행되며 -0.7 V vs. Ag/AgCl/4 N-HCl에서 N-N=O기의 선택적인 4전자 환원반응에 의해 N-N$H_2$가 생성됨을 확인하였다.

• 대한바이러스학회지
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• 제27권1호
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• pp.87-95
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• 1997

Methanol and/or boiling water extraction of 201 natural products and subsequent MTT assay using MT-4 cell line was carried out to screen the anti-HIV-1 activity. Among 97 methanol extracts, 7 extracts from Chrysanthemi Indicium Flos, Magnoliae Cortex, Machili Cortex, Reynoutriae Rhizoma, Lithospermi Radix, Agastachis Herba, and Chaenomelis Fructus showed anti-HIV-1 activity and their SI value were 2.25 to 5.77. In addition, among 119 boiling water extracts, 10 extracts from Lonicerae Caulis et Foloium, Elsholtziae Herba, Leonuri Herba, Portulacae Herba, Schizonepetae Herba, Curcumae Rhizoma, Amomi Cardamomi Fructus, Cirsii Radix et Herba, Carpesii Herba, and Siegesbeckiae Herba showed anti-HIV-1 activity and their SI value were 1.30 to 7.64. Methanol extracts of above seven natural products were fractionated and the anti-HIV-1 activity of each fraction was examined. Extraction was carried out with hexane, chloroform, butanol, and water to trace active anti-HIV-1 componets. As a result, the water fraction of Magnoliae Cortex, Machili Cortex, Reynoutriae Rhizoma, Agastachis Herba, Chaenomelis Fructus and the butanol fraction of Chrysanthemi Indicium Flos, Reynoutriae Rhizoma showed anti-HIV-1 activity and their SI value were 1.40 to 8.02. We could reach a conclusion that studies to trace the anti-HIV-1 active component of each natural products in further fractionation and to identify its structure by Infrared spectroscopy, NMR spectroscopy and gel permeation chromatography were needed.

• Applied Biological Chemistry
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• 제45권3호
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• pp.168-172
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• 2002

한국산 옻나무의 메탄올추출물과 유기용매 분획의 항산화 효과를 조사하였다. 항산화 효과는 수소공여 억제능, 과산화지질 형성 억제능, xanthine oxidase저해 활성과 아질산염 소거능으로 측정한 결과, ethyl acetate 분획이 가장 뛰어난 효과를 나타내었다. 이 분획의 생리활성 물질을 분리하기 위하여, rotatory locular counter current chromatography(RLCCC), Sephadex LH-20 column chromatograpy와 HPLC등의 방법을 사용하여, 1,2,3-trihydroxybenzene, methyl 3,4,5-trihydroxybenzoat와 3,4,5-trihydroxybenzoic acid를 분리하였다. 이중 methyl 3,4,5-trihydroxybenzoate는 butylated hydroxyanisole(BHA)이나 butylated hydroxytoluene(BHT)보다도 강한 항산화 효과를 나타내었다. 그러나 세포독성 실험에서는 이들 물질들이 암세포주에 대하여 활성이 높지 않았다. 이상의 결과로 methyl 3,4,5-trihydroxybenzoate은 뛰어난 항산화 효과를 지닌 식품첨가물로서의 이용 가능성을 제시할 수 있다.