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토양으로 부터 분리한 방선균 KYR 26균주가 생산하는 항생물질의 분리 정제
유진철,방희재,심정보,이은하,김형근,김성준,김영호,이정준 조선대학교 부설 유전생물공학연구소 1993 유전생물공학연구지 Vol.3 No.-
1. 작용 범위가 넓고 부작용이 적은 의약용 항생물질을 screening하기 위한 목적으로 전남 지역 토양시료로 부터 방선균을 분리하여 항균활성이 우수한 방선균을 선별하여 KYR 26이라 명명 하였다. 2. KYR 26균주의 최적 배양배지는 Modified SMM배지 이었으며, 항생물질 생산은 배양 3일째에 최대의 수율을 나타내었다. 3. KYR 26균주가 생산하는 항생 물질은 Ion exchange chromatography, Adsorption chromatography, LH-20 coluom chromatography, Precipitation과 Silica column chromatography 과정을 통하여 항생물질 A, B를 분리정제 하였다. 4. 분리 정제된 항생 물질 A와 B는 TLC에서 각각 Rf치 0.53, 0.48에서 단일 spot로 나타났고,HPLC에서 항생물질 A와B는 각각 2.94, 7.66의 retention time을 보였으며 모두 98%의 순도를 보였다. 5. 분리 정제된 항생 물질 A와 B의 UV spectrum을 조사한 결과 항생물질 A,B는 모두 270nm에서 최대흡수 파장을 나타내었다. 현재 최종 분리 정재된 항생물질 A와 B의 구조 규명을 위하여 IR, ^1H-NMR, ^3C-NMR, Mass spectroscopy에 관한 실험이 진행중에 있다. Soil Actinomycetes were isolated from soil samples which were collected from the 100 sites of Cheon-nam areas. Isolated strains were tested for the production of antibiotics in various fermentation media using 4 strains of antibiotic sensitive test microorganisms. One strain which has excellent antibacterial activities was selected, and named to KYR 26. From culture broth of KYR 26 strain, two antibiotics were obtained through Ion Exchange Chromatography, Adsorption Chromatography, Gel Filtration Chromatography, Silica gel column chromatography and Crystalization. The finally purified two antibiotics exhibited single spot on the TLC chromatogram,and about 98 % purity on the profile of HPLC Chromatogram. Two antibiotics exhibited Rf 0.53 and 0.48 each on TLC with developing solvent system (MeOH:CHCl_3). The UV spectrum of isolated antibiotics exhibited a characteristic absorption maximum at 270nm.
6극 전자석 전자교반 레오캐스팅에 따른 A356의 조직적 /기계적 영향분석
김백규,노중석,방희재,허민,박진하,전충환 한국주조공학회 2020 한국주조공학회지 Vol.40 No.3
Rheo-diecasting with stirring has been used in many material industries. As the 4th Industrial Revolution approaches the world, eco-friendly high-strength and light-weight materials become more important. Casting methods have been studied and used for aluminum-alloy automobile parts. This study carried out the effect analysis of the micro-structure and mechanical properties, such as yield/ultimate tensile strength, elongation, and hardness, of A356 using the 6-pole EMS (electro-magnetic stirring) casting process with a high electromagnetic force. As a result, the hardness and elongation of the A356 after T6 heat-treatment show a significant improvement, respectively, by 20% and 50%.
인체유암세포주 MCF-7의 형태변화와 증식에 영향을 주는 항암활성물질, MCS-202
이성우,김세은,김영호,김항섭,방희재,김환묵,이정준 한국산업미생물학회 1993 한국미생물·생명공학회지 Vol.21 No.6
미생물 대사산물로부터 인체유암세포주 MCF-7의 형태변화와 세포독성을 나타내는 화합물을 탐색하는 중에 Streptomyces sp. 1365 균주를 선별하였으며, 이 균주를 배양하여 활성물질 MCS-202를 분리하였다. 각종 spectroscopic data에 의한 구조분석의 결과 이 화합물은 resistomycin 으로 동정하였다. 이 화합물은 50 pg/㎖부터 50 ng/㎖의 농도에서 MCF-7 세포의 형태변화를 유도하였으며, 50 ng/㎖ 이상의 농도에서는 세포독성을 나타내었다. 이 화합물에 대하여 NCI의 tumor panel 중 9종류의 인체암세포주에 대한 항암활성을 조사하여 본 결과 0.1∼0.3㎍/㎖의 농도에서 거의 모든 세포에 대하여 강한 세포독성을 나타내었다. In the course of screening for microbial metabolites employing human cancer cell line, we identified a mycelial extract of Streptomyces sp. 1365, which are effective on growth inhibition and morphological change of MCF-7, human breast cancer cell line. By repeated column chromatography and recrystallization process, yellow needle crystals were obtained as an active compound and identified as resistomycin by spectral analysis. Morphological change of the MCF-7 cell could be observed by treatment of resistomycin at concentration between 50 ng/㎖ and 50 pg/㎖ and showed cytotoxic effect at the concentration of more than 50 ng/㎖. This compound also showed strong cytotoxic effect on proliferation of human tumor cell lines, UACC-62, NCI-H23, SNB-19, SF-539, PC-3, SW620, KM12, UO-31, and ACHN.
김항섭,김세은,이성우,방희재,김영호,이정준,Kim, Hang-Sub,Kim, Se-Eun,Lee, Sung-Woo,Bang, Hee-Jae,Kim, Young-Ho,Lee, Jung-Joon 한국미생물 · 생명공학회 1994 한국미생물·생명공학회지 Vol.22 No.4
Three more unusual macrolides in addition to concnamycin B were isolated from the mycelium of Streptomyces sp. strain Bal6. These four compounds showed a potent cytotoxity to hunian cancer cell lines, SNU-1 (stomach cancer cell line), SNU-354 (liver cancer cell line), MCF- 7 (breast cancer cell line) and KB-3-1 (oral epidermoid carcinoma cell line). Interestingly, these compounds confered slight differential cytotoxity on RHEK-1, a human epidermal keratinocyte cell line immotalized by AD12-SV40 hybrid virus and RHEK-1/pSV$_{2}$ ras which was resulted from H-ras transfomation of RHEK-1. These compounds were determined to be concanamycin A, conca- namycin E and 0-methyl concanamycin B by NMR and other spectral analysis.
3-(치환) 테트라조일메칠세파로스포린의 합성과 생리활성
고옥현(Ok Hyun Ko),김영수(Young Soo Kim),고봉석(Bong Suk Ko),이재영(Jae Young Lee),하재천(Jai Chun Ha),방희재(Hee Jae Bang),유진철(Jin Cheol Yoo),강형룡(Hyung Ryong Kang) 대한약학회 1998 약학회지 Vol.42 No.1
For the development of new cephalosporin antibiotics with aminothiazolcarboxymethylethoxyimino moiety on the C-7 position and tetrazolymethyl moiety on the C-3 position of cephem ring, 70-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[5-(substituted)tetrazol-2-yl]methyl-3-cephem-4-carboxylic acids(28-35) were synthesized. These compounds were tested for antimicrobial activity in vitro against Gram(+) and Gram(-) bacteria. They showed remarkable antibacterial activity against Escherichia coli AB 1157, Escherichia coli AB 0111, Escherichia coli BE 1186, Micrococcus luteus ATCC 9341, Salmonella typhimurium TV 119, Salmonella typhimurium SL 1102, Staphylococcus aureus IFO 12732, Staphylococcus aureus R-209, but these compounds were not active against Pseudomonas aeruginosa N-10.