DSpace at EWHA: Studies on Isolation, Identification, and Biological Activities of Antifungal Antibiotics produced by Streptomyces willmorei EMP22, against Cryptococcus laurentii

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DSpace at EWHA일반대학원 생명·약학부 Theses_Master

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DC Field	Value	Language
dc.contributor.author	은소영	-
dc.creator	은소영	-
dc.date.accessioned	2016-08-26T10:08:45Z	-
dc.date.available	2016-08-26T10:08:45Z	-
dc.date.issued	1997	-
dc.identifier.other	OAK-000000023402	-
dc.identifier.uri	https://dspace.ewha.ac.kr/handle/2015.oak/198681	-
dc.identifier.uri	http://dcollection.ewha.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000023402	-
dc.description.abstract	The aim of our current research is to isolate and purify the antifungal compounds produced by Streptomyces willlmorei EMP22 *isolated from soil and to carry out the analysis of their structures in order to develop antifungal antibiotics functioning as a cell wall synthesis-inhibitor without influencing in human body. Antibiotic- producing strain, EMP22 was identified as S. willlmorei(Park, 1995). Antifungal compounds were isolated from the culture filtrate of the strain by use of Amberlite XAD-2(macroreticular resin) column chromatography, Preparative TLC, Sephadex LH-20(gel filtration) column chromatography, and Preparative HPLC. The main principle of the several compounds showing antifungal activities was fractionated and its structure was determined by instrumental analysis such as UV, NMR, and MS techniques. The compound was identified to be tunicamycin Ⅶ, a carcinogenic, antiviral, and antifungal antibiotic produced by S. lysosuperificus. However, this is the first report that tunicamycin Ⅶ has an antifungal activity against Cryptococcus laurentii NCYC 139.;이번 연구의 목적은 인체에는 해를 주지 않으면서 진균의 세포벽 합성을 저해하는 항진균성 항생물질을 개발하는 것이었다. 먼저, 토양에서 분리하여 동정한 Streptomyces willmorei EMP22가 생산하는 항진균성 항생 물질을 분리정제하고, 그것의 구조분석을 시행하였다. 항진균성 물질들은 배양 상등액으로부터 Amberlite XAD-2(macroreticular resin) column chromatography, 분취용 TLC, Sephadex LH-20(gel filtration) chromatography, 그리고 분취용 HPLC를 이용하여 분리되었다. 항진균 활성을 보이는 여러 peaks들 가운데 주요한 하나의 peak을 골라 그것의 fraction을 받아서, UV, NMR, MS techniques을 이용한 구조분석을 수행하였다. 그 결과, 이 물질은 발암물질이며, 항바이러스제이며, 항진균물질인 Tunicamycin Ⅶ 과 구조가 동일하였다. 이 항진균물질은 S. lysosuperificus에 의해 생산되는 것으로 알려져왔다(Tamura, 1982). 그러나 Tunicamycin이 Cryptococcus laurentii NCYC 139 에 대한 항진균 활성을 갖는다는 연구결과는 본 논문이 처음 보고하는 것이다. 본 연구는 C. laurentii 이외에 Aspergillus flavus, A. Penicillium citrinum 에 대한 활성도 조사하였다.	-
dc.description.tableofcontents	Contents Abstract = 1 Introduction = 2 1. Antibiotics : General Explanation = 2 2. Search for new antibiotics = 2 3. Steps toward commercial production = 4 4. Isolation of antifungal antibiotics = 5 5. Discovery of Tunicamycin = 5 Materials and Methods = 10 1. Test Organism = 10 1.1. Cultural Condition of the test organism = 10 2. Cultural Conditions for Antibiotic-Producing Micro-organisms = 10 2.1. Medial Constitutions = 10 2.1.1. Slant Medium for Strain-Preservation = 10 2.1.2. Medium for Antibiotic-Production = 11 2.2. Cultivation = 11 2.3. Harvest = 12 3. Antifungal Activity Assay = 12 4. Isolation and Purification = 12 4.1. Studies on Solvent-Extraction = 12 4.2. Amberlite XAD resin-adsorption test = 13 4.3. Amberlite XAD-2 Column Chromatography = 13 4.4. Preparative TLC = 13 4.5. Sephadex LH-20 Column Chromatography = 14 4.6. Preparative HPLC = 14 4.7. Analytical HPLC = 15 5. Studies on Physico-chemical Properties and Identification = 15 5.1. UV Spectrometry = 15 5.2. Solubility test = 15 5.3. Color reaction = 16 5.3.1. Anisaldehyde-H_(2)SO_(4) test = 16 5.3.2. Dragendorff's test = 16 5.3.3. Ferric Chloride test = 16 5.3.4. I_(2) vapour test = 17 5.3.5. Ninhydrin test = 17 5.3.6. Periodic acid-benzidine test = 17 5.3.7. Phosphomolybdic acid test = 17 5.3.8. Potassium Permanganate test = 17 5.3.9. Sakaguchi test (1-Naphthol-Bromine) = 18 5.4. pH Stability test = 18 5.5. Thermostability test = 18 6. Identification of EMP22-1 = 18 6.1 Mass Spectrometry = 18 6.2. NMR Spectroscopy = 18 7. Biological Activities of EMP22-1 = 19 7.1. Morphological Abnormality = 19 7.2. Antimicrobial Spectrum = 19 Results and Discussion = 20 1. Isolation = 20 1.1. Studies on Solvent-Extraction = 20 1.2. Amberlite XAD resin-adsorption test = 20 1.3. Amberlite XAD-2 Column Chromatography = 21 1.4. Preparative TLC = 21 1.5. Sephadex LH-20 Column Chromatography = 21 1.6. Preparative HPLC = 22 1.7. Analytical HPLC = 22 2. Studies on Physico-chemical Properties = 22 2.1. UV spectrometry = 22 2.2. Solubility test = 30 2.3. Color Reaction = 30 2.3.1. Anisaldehyde-H_(2)SO_(4) test = 30 2.3.2. Dragendorff's reagent test = 30 2.3.3. Ferric Chloride test = 30 2.3.4. I_(2) vapour test = 30 2.3.5. Ninhydrin test = 31 2.3.6. Periodic acid-Benzidine test = 31 2.3.7. Phosphomolybdic acid test = 31 2.3.8. Potassium Permanganate test = 31 2.3.9. Sakaguchi test (1-Naphthol-Bromine) = 31 2.4. pH Stability test = 31 2.5. Thermostability test = 32 3. Identification of the Antifungal Compound, EMP22-1 = 32 4. Biological Activities of Tunicamycin = 33 4.1. Morphological Abnormality induced by EMP22-1 (Tunicamycin Ⅶ) = 33 4.2. Antimicrobial Spectrum of Tunicamycin = 34 References = 52 초록 = 58	-
dc.format	application/pdf	-
dc.format.extent	2322812 bytes	-
dc.language	eng	-
dc.publisher	Graduate School of Ewha Womans University	-
dc.subject	Streptomyces	-
dc.subject	EMP22	-
dc.subject	Cryptococcus laurentii	-
dc.subject	Isolation	-
dc.title	Studies on Isolation, Identification, and Biological Activities of Antifungal Antibiotics produced by Streptomyces willmorei EMP22, against Cryptococcus laurentii	-
dc.type	Master's Thesis	-
dc.title.subtitle	Streptomyces willmorei EMP22가 생산하는 Cryptococcus laurentii에 대한 항진균성 항생물질의 분리정제, 구조동정 및 생물학적 활성 연구	-
dc.format.page	v, 58 p.	-
dc.identifier.thesisdegree	Master	-
dc.identifier.major	대학원 생물과학과	-
dc.date.awarded	1998. 2	-