항진균성 항생물질을 생산하는 Bacillus sp. LAM 97-44의 분리 및 동정

이노운,김천석,도재호,정인찬,이현우,이동희,Lee, No-Woon,Kim, Cheon-Suk,Do, Jae-Ho,Jung, In-Chan,Lee, Hyean-Woo,Yi, Dong-Heui 한국응용생명화학회 1998 Applied Biological Chemistry (Appl Biol Chem) Vol.33 No.2

항진균제를 개발하기 위하여 토양으로부터 700주의 세균, 방선균, 곰팡이를 분리하였으며, 이들 배양액을 조사한 결과 Candida albicans의 성장을 억제시킬 수 있는 LAM 97-44 균주를 항진균성 항생물질 생산균으로 선발하였다. 분리균주를 동정하기 위해 생산균 LAM 97-44 균주의 형태학적 특성, 배양학적 특성, 생리학적 특성, 균체성분 등을 조사하였다. LAM 97-44 균주는 $2{\sim}3{\times}1{\sim}1.5\;{\mu}m$ 크기의 포자를 형성하는 간균이며, 포자의 형태는 ellipsoid 형 이었다. LAM 97-44 균주는 arabinose, cellulose, xylose를 이용하지 못하였으나 fructose, glucose, glycerol, maltose, raffinose 등은 이용하는 것으로 나타났다. 지방산 분석결과는 iso-와 anteiso-인 성분으로 구성되어 있었으며, menaquinone은 Bacillus속 세균의 전형적인 isoprenoid 사슬이 7개인 menaquinone(MK-7)이었다. 이러한 결과를 종합하면 LAM 97-44 균주는 Bacillus subtilis와 유사한 균으로 동정되었다. In order to develop an effective antifungal antibiotics, over 700 isolates of bacteria, mold and actinomytes were screened from soil, and LAM 97-44 were selected as a strain producing the strong antifungal antibiotics against Candida albicans. Morphological, cultural and physiological characteristics of LAM 97-44 were investigated for the indentification. The cell size of LAM 97-44 was $2{\sim}3{\times}1{\sim}1.5\;{\mu}m$, and the shape of spore was of ellipsoidal. As a carbon source, LAM 97-44 utilized fructose, glucose, glycerol, maltose and raffinose but did not utilize arabinose, cellulose and xylose. The fatty acids of the cells included various iso-type and anteiso-type. Conclusively, the strain LAM 97-44 was proved to be Bacillus subtilis.

항진균성 항생물질를 생산하는 Bacillus sp. LAM 97-44 의 분리 및 동정

이노운,김천석,도재호,정인찬,이현우,이동희 ( No Woon Lee,Cheon Suk Kim,Jae Ho Do,In Chan Jung,Hyean Woo Lee,Dong Heui Yi ) 한국응용생명화학회 1998 Applied Biological Chemistry (Appl Biol Chem) Vol.41 No.3

In order to develop an effective antifungal antibioltics over 700 isolates of bacteria, mold and actinomytes were screened from soil., and LAM 97-44 were selected as a strain producing the strong antifungal antibiotics against Candida albicans. Morphological, cultural and physiological characteristics fo LAM 97-44 were investigated for the indentification. The cell size of LAM 97-44 was 2∼3×1∼1.5 ㎛, and the shape of spore was of ellipsoidal. As a carbon source, LAM 97-44 utilized fructose, glucose, glycerol, maltose and raffinose but did not utilize arabinose, cellulose and xylose. The fatty acids of the cells included various iso-type and antciso-type. Conclusively, the strain LAM 97-44 was proved to be Bacillus subtilis.

Bacillus subtilis LAM 97-44가 생산하는 항진균성 항생물질의 정제 및 특성

이노운,권태종,이동희,Lee, No-Woon,Kwon, Tae-Jong,Yi, Dong-Heui 한국응용생명화학회 2003 한국농화학회지 Vol.46 No.2

병원에서 분리한 azole계 항진균성 항생물질에 대한 내성을 가지고 있는 Candida albicans에 대해 강한 활성을 가지는 항진균성 물질을 Bacillus subtilis LAM 97-44의 배양액으로부터 분리 정제한 후 그 특성을 조사하였다. 원심분리한 배양상등액을 butanol 추출, Diaion HP-20과 Dowex-50 adsorption chromatography, silica gel flash chromatography와 HPLC로 정제하였고 TLC와 HPLC로 확인하여 그 물질을 LAM-44A라 명명하였다. LAM-44A는 pH와 열에 매우 안정하였으며 Candida sp., Cryptococcus sp. 등에 대해 강한 활성을 나타낸 반면에 독성은 매우 적었다. 분리한 물질은 273 m에서 최대흡광도를 가진 융점 $202^{\circ}C$의 무색분말이었으며 ninhydrin 반응결과 음성이었고 $^1H-NMR$, $^{12}C-NMR$, IR spectrum, 원소분석 등의 결과로 볼 때 분자량 282의 $C_{14}H_{34}O_5$의 화학식을 가진 물질로 동정되었다. A novel antifungal antibiotic for azole-resistant Candida albicans was purified from the culture broth of Bacillus subtilis LAM 97-44 by butanol extraction, Diaion HP-20 and Dowex-50 adsorption chromatography, silica gel flash chromatography followed by HPLC and designated LAM-44A. LAM-44A was stable for 60 min at $100^{\circ}C$, and pH range from 2 to 10. MIC values were observed at $0.5-3.5\;{\mu}g/ml$ against various Candida albicans strains. The antibiotic showed no cytotoxicity for S180, MKN-45, P388, HeLa and 373 at the concentration of 1 mg/ml. LAM-f4A was colorless powder soluble in water, methanol, ethanol, butanol and negative to ninhydrin reaction. The antibiotic had maximum absorption at 273 nm in methanol, and melting point was $202^{\circ}C$. The molecular weight and formula were determined to be 282 and $C_{14}H_{34}O_5$ by $^1H-NMR,\;^{13}C-NMR$, IR spectrum and elemental analysis.

Streptomyces aureofaciens 동결건조후 재수화 방법이 생존도에 미치는 영향

이노운(No Woon Lee),이현우(Hyean Woo Lee),이동희(Dong Heui Yi) 한국응용생명화학회 1995 Applied Biological Chemistry (Appl Biol Chem) Vol.38 No.6

In order to examine the effect of rehydration methods on viability after freeze-drying of Streptomyces aureofaciens, we labeled the DNA of S. aureofaciens with ³H-adenine. Extracellular radioactivity levels appeared to be high in the rehydrated solutions after freeze-drying than freezing-thawing. In effects of rehydration after freeze-drying, the viability of the cell appeared about 20% in case of with admitting air at single ampule, but that of which appeared about 91% in case of without admitting air at double ampule. Thus, S. aureofaciens cells were damaged during the process of rehydration after freeze-drying.

[화학] 역상 고속 액체크로마토그래피를 이용한 인삼 사포닌의 분석

정승일(Seung-Il Jeong),김천석(Choen-Suk Kim),이노운(No-Woon Lee),최강주(Kang-Ju Choi),이용구(Yong-Gu Lee),김일광(Il-Kwang Kim) 한국분석과학회 1998 분석과학 Vol.11 No.6

올리브유를 첨가한 빵 반죽의 리올로지 특성

임선희(Sun-Heui Lim),김석영(Seok-Young Kim),이노운(No-Woon Lee),이치호(Chi-Ho Lee),이시경(Si-Kyung Lee) 한국식품과학회 2004 한국식품과학회지 Vol.36 No.5

수용성 항진균성 항생물질을 생산하는 방선균 LAM-593의 분리 및 물질생산조건

이동희,김중배,이노운 建國大學校 附設 産業技術硏究所 1993 논문집 Vol.18 No.-

토양에서 분리 선별한 수용성 항진균성 물질을 생산하는 Streptomyces속 균주 LAM-593에 의한 항진균성 물질의 생산조건을 검토한 결과는 다음과 같다. 물질생산을 위한 탄소원으로는 soluble starch가, 질소원으로는 soybean meal과 peptone이 가장 우수하였으며 glycerol의 첨가로 물질 생성량이 현저히 증가하였으나 인산염에 의해서는 저해되었다. 생산 최적온도와 초발 pH는 각각 30℃와 9.5였으며, soluble starch 1.0%, glycerol 1.0%, soybean meal 0.3%, peptone 0.1%, MgSO₄·7HO 0.05%, NaCI 0.05%, silicone oil 0.01% 조성의 배지(pH 9.5) 2.5L를 5L fermenter에 넣고 전배양한 배양액을 3% 수준으로 접종하여 통기량 0.9vvm, 교반속도 200rpm으로 30℃ 에서 2일간 배양하였을 때 비지내의 항진균성 물질의 축적량이 최대에 달하였다. Cultural conditions for production of water soluble antifungal antibiotic by Streptomyces sp, strain LAM-593, isolated from soil, were studied. For the antibiotic production, soluble starch and soybean meal was favorable as carbon and nitrogen source, respectively. The antibiotic production was stimulated by the addition of Mg? and glycerol. Optimal temperature and initial pH for production of the antibiotic was 30℃ and pH 9.5, respectively. Accumlation of the antibiotic in th culture broth reached at maximum level after 2 days cultivation under aeration rate 0.9 vvm and impeller speed 200rpm in 5L-fermenter with 3L of medium containing soluble starch 1.0%, glycerol 1.0%, soybean meal 0.3%, peptone 0.1%, MgSO₄·7H₄O 0.05% and Nacl 0.05%.

세포응집물질을 생산하는 Aspergillus sp. LAM 94-142의 특성 및 물질생산 조건

이동희,이노운,이현우 건국대학교 산업기술연구원 1996 건국기술연구논문지 Vol.21 No.-

토양에서 분리한 세포응집물질을 생산하는 균주 LAM 94-142는 형태, 배양 및 생리학적 특성을 조사한 결과 Aspergillus parasiticus 또는 그 근연균으로 동정되었으며 물질을 생산하기 위한 배양최적 조건을 검토한 결과는 다음과 같다. 최적 탄소원과 질소원은 각각 1.5% glucose와 0.35% yeast extract였으며 Ca²+을 첨가하였을 때 물질생산이 현저히 증가되었으며 K+, Cu²+, Fe²+ 등의 금속이온은 물질 생성을 저해하였다. 또 인산의 K염은 물질 생합성을 촉진하였으나 ammonium염은 크게 저해하였다. 그리고 최적 초발 pH와 온도는 pH7과 30℃였으며 4일간 배양하였을 때 물질 생산량이 최대였다. A strain of mold, LAM 94-142, capable of producing cell aggregation factor was isolated from soil and identified as Aspergillus parasiticus or its related strain on the morphological, cultural and physiological characteristics. For production of the cell aggregation factor, glucose and yeast extract was favorable as carbon and nitrogen source, respectively. The factor production was increased by the addition of Ca²+ and potassium phosphates but inhibited by K+, Cu²+, Fe²+ and ammonium phosphates. Optimal initial pH and temperature for the factor production was pH 7 and 30℃, respectively. The production of the factor reached at maximum after 4 days in the medium containing 1.5% glucose, 0.35% yeast extract, 0.07% CaCl₂, and 0.05% KH₂PO₄.

수용성 항진균성 항생물질 생산을 위한 Bacillus sp. LAM 97-44의 배양조건

이동희,이노운 건국대학교 산업기술연구원 2000 건국기술연구논문지 Vol.25 No.-

A strain of Bacillus sp. LAM 97-44, capable of producing antifungal antibiotic was isolated and its cultural conditions for production of the substance was investigated. For production of the water-soluble antifungal antibiotic, glucose, glycerol, malt extract and ammonium phosphate were favorable as carbon and nitrogen source. The antibiotic production was increased by the addition of Mg2+ and Fe2+. Optimal initial pH and temperature was pH 7.0 and 30℃, respectively. The production of the antibiotic reached at maximum after 5 days in the medium containing 1.3% glucose, 0.4% glycerol, 0.1% malt extract, 0.2% NH4H2PO4, 0.1% K2HPO4, 0.03% MgSO4·7H2O and 0.03% FeSO4·7H2O.

Bacillus subtilis LAM 97-44가 생산하는 항진균성 항생물질의 정제 및 특성

이동희,이노운,권태종 한국농화학회 2003 Applied Biological Chemistry (Appl Biol Chem) Vol.46 No.2

병원에서 분리한 azole계 항진균성 항생물질에 대한 내성을 가지고 있는 Candida albicans에 대해 강한 활성을 가지는 항진균성 물질을 Bacillus subtilis LAM 97-44의 배양액으로부터 분리 정제한 후 그 특성을 조사하였다. 원심분리한 배양상등액을 butanol 추출, Diaion HP-20 과 Dowex-50 adsorption chromatography, silica gel flash chromatography와 HPLC로 정제하였고 TLC와 HPLC로 확인하여 그 물질을 LAM-44A라 명명하였다. LAM-44A는 pH와 열에 매우 안정하였으며 Candida sp., Cryptococcus sp. 등에 대해 강한 활성을 나타낸 반면에 독성은 매우 적었다. 분리한 물질은 273 nm에서 최대흡광도를 가진 융점 202˚C의 무색분말이었으며 ninhydrin 반응결과 음성이었고 ^1H-NMR, ^13C-NMR, IR spectrum, 원소분석 등의 결과로 볼 때 분자량 282의 C_14H_34O_5의 화학식을 가진 물질로 동정되었다. A novel antifungal antibiotic for azole-resistant Candida albicans was purified from the culture broth of Bacillus subtilis LAM 97-44 by butanol extraction, Diaion HP-20 and Dowex-50 adsorption chromatography, silica gel flash chromatography followed by HPLC and designated LAM-44A. LAM-44A was stable for 60 min at l00˚C, and pH range from 2 to 10. MIC values were observed at 0.5-3.5 ㎍/㎖ against various Candida albicans strains. The antibiotic showed no cytotoxicity for S180, MKN-45, P388, HeLa and 3T3 at the concentration of 1 ㎎/㎖. LAM-44A was colorless powder soluble in water, methanol, ethanol, butanol and negative to ninhydrin reaction. The antibiotic had maximum absorption at 273 nm in methanol, and melting point was 202˚C. The molecular weight and formula were determined to be 282 and C_14H_34O_5 by ^1H-NMR, ^13C-NMR, IR spectrum and elemental analysis.