Aminopeptidase M 저해제인 Valistatin과 des-Asp^4-Amastatin을 생산하는 방선균 SL20209의 특성 및 동정

고학룡,전효곤,정명철,서현효,김홍중,박용하,고영희 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1

Aminopeptidase M 저해제인 valistatin과 des-asp^4-amastatin을 생산하는 방선균 분리주 SL20209의 특성을 조사하고 수리동정을 실시하였다. 형태적, 배양학적 및 생리적 특성으로 부터 SL20209는 방성균 중에서도 Streptomyces 속에 속하는 것으로 나타났으며, 43개의 분류단위 특성으로 TAXON program을 이용하여 수리동정한 결과, 주군집 29의 Streptomyces 중 Streptomyces griseoplanus와 가장 근접한 것으로 나타났다. 따라서, 분리주 SL20209를 S. griseoplanus SL20209로 동정하였다. Characterization and numerical identification were carried out for an actionmycetes SL20209. Morphological, cultural and physiological properties of SL20209 which produced valistatin and des-asp^4-amastatin as inhibitors of aminopeptidase M were evaluated. The isolate was identified to be the genus of Streptomyces. Fourty-three taxonomic units were analysed by using a TAXON program. The isolate was classified into the major cluster 29 of Streptomyces and best-matched to Streptomyces griseoplanus.

미생물 모방대사를 이용한 천연물의 생물전환

고학룡,안순철,Ko, Hack-Ryong,Ahn, Soon-Cheol 한국해양바이오학회 2007 한국해양바이오학회지 Vol.2 No.1

This aims to review natural products transformed by mimic intestinal metabolisms with microorganisms and hydrolytic enzymes, which exhibit enforced biological activity, higher extraction yield and identification of active components. In the process, transformation to the smaller active compounds with enzymes and microbes mimics the pharmacological action of natural products by intestinal bacteria. In order to establish conditions for the fermentation and enzyme reaction, it is required to choose several natural products for biotransformation and investigate the optimal conditions for the fermentation or the enzyme reaction such as composition, temperature, pH, inoculum, and cultivation time. It is expected an increase of the internal absorption of the active materials without regard to the intestinal microbes or its ability through biosynthesis of the active materials by the microbes and enzymes. And this techniques can be applied to biotransformation of natural products such as sesaminol, resveratrol, 1-deoxy nojirimycin, naringenin, quercetin, and baicalin and to the metabolism study using the animal model.

Streptomyces griseoplanus SL20209에 의한 Aminopeptidase M 저해제의 생산 조건

고학룡,전효곤,성낙계,고영희 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.3

생산용 배지(GSS) 100 ㎖을 포함하는 500㎖ 삼각 플라스크에서 Streptomyces griseoplanus SL20209를 28℃, 3일간 배양했을 때 aminopeptidase M(AP-M) 저해제 생산이 최대에 도달하였으며, 그 이후 pH가 알카리성으로 변하면서 다소 감소하였다. Arabinose, xylose, mannose 및 soluble starch가 저해제 생산에 좋은 탄소원인 반면, glucose는 균생육에는 좋으나 저해제 생산은 강하게 저해하였다. Soybean meal, fish meal, gluten meal 및 peanut powder와 같은 천연 유기질소원이 저해제 생산에 좋은 질소원이었으며, soytone이나 peptone 및 NH_4Cl과 NH_4NO_3와 같은 무기질소원이 함유된 배지에서는 저해제 생산이 빈약하였다. 배지내에 각각 0.5%(w/v)의 yeast extract난 0.05%의 K_2HPO_4의 첨가는 균생육을 촉진시켜 저해제 생산을 증가시켰으며, CaCo_3(0.3%)와 zeolite(0.5%)도 저해제 생산 증가의 효과를 나타내었다. 저해제 생산을 위한 최적 배양온도 및 초기 pH 범위는 각각 28℃와 6.0~7.0이었다. Arabinose(ASY 배지) 또는 soluble starch(SSY 배지) 3%, SBM 2.5%, yeast extract 0.5%, K_2HPO_4 0.05%, CaCO_3 0.1% 및 zeolite 0.3%(pH 6.8)의 조성의 두 개선된 배지에서 초기의 GSS 배지에서 보다 저해제 생산이 1.8배 증가되었다. Maximum amount of the aminopeptidase M inhibitors produced by Streptomyces griseoplanus SL20209 in 500 ml-Erlenmeyer flask was accumulated after cultivation for 3 days at 28℃, thereafter the amount of inhibitors decreased slowly with a pH change to alkaline. Arabinose, xylose, mannose and soluble starch were good carbon sources for the production of the inhibitors. On the other hand, glucose was only good for the cell growth but potently inhibited the production of inhibitors. Natural organic nitrogen sources such as soybean meal, fish, meal, gluten meal and peanut powder were good for the production of inhibitors, however, soytone, peptone and inorganic nitrogens such as NH_4Cl and NH_4NO_3 were poor. Inclusion of yeast extract (0.5%, w/v) or K_2HPO_4 (0.05%) into the production medium increased the production of inhibitors by accelerating cell growth. The production of inhibitors was slightly increased on the medium containing CaCo_3(0.3%) and zeolite (0.5%), respectively. Optimal temperature and initial pH range for the production of inhibitors were determined to be 28℃ and 6.0~7.0, respectively. Employing two improved production media consisting of 3% arabinose or soluble starch, 2.5% soybean meal, 0.5% yeast extract, 0.05% K_HPO_4 0.1% CaCO_3 and 0.3% zeolite (pH 6.8), 1,8-fold increase in the amount of inhibitors was achieved, comparing with the basal medium used.

일반발표 - 생화학 : Streptomyces sp . 20209 로부터 Aminopeptidase M 저해제의 분리 및 특성

고학룡,정명철,전효곤,고영희,성낙계 한국토양비료학회 1993 한국토양비료학회 학술발표회 Vol.1993 No.50

생화학 및 분자생물학 : Streptomyces sp . 20209 로 부터 Aminopeptidase M 저해제의 분리 및 특성

고학룡,정명철,전효곤,고영희,성낙계 한국응용생명화학회(구 한국농화학회) 1993 한국응용생명화학회 학술발표회 Vol.1993 No.-

섬유질 가수분해물로부터 효율적인 Ethanol 생산균주의 분리

고학룡,문종상,심기환,성낙계 한국산업미생물학회 1991 한국미생물·생명공학회지 Vol.19 No.4

섬유성 가수분해물로부터 효율적으로 ethanol을 생산하는 세 가지 균주를 밀기울 당화액에서의 집적 배양에 의해 토양에서 분리하였다. 효모인 KM-09와 KM-402 및 세균인 HG-225의 생리학적 및 생화학적 특성은 각각 Candida sp. 및 Klebsiella sp.과 거의 유사하였다. KM-09와 HG-225 균주는 발효당으로 xylose와 cellobiose를 이용하였고 HG-225는 발효시 넓은 당 이용성을 가졌다. KM-09, KM-402 및 HG-225의 최적 생육 pH 및 온도는 각각 5.8, 5.6 및 6.8 그리고 32℃, 30℃ 및 38℃였다. 분리균에 의한 밀기울 당화액에서 ethanol 발효 동안 최대 ethanol 생산온도는 최적생육온도보다 다소 높았으며, 0.2% MgSO_4의 첨가는 ethanol 생산성을 향상시켰다. 세 가지 균주 중 KM-09의 ethanol 함량이 약 2.3%(v/v)로서 가장 높았고 HG-225의 ethanol 생산속도가 가장 빨랐으며 이때 최대 생산성은 배양 4일 후였다. KM-09와 HG-225는 4%(w/v) xylose로부터 각각 1.42 및 1.05%(v/v)의 ethanol을 생산하였으나 균 증식속도는 glucose에 비해 매우 느린 것으로 나타났다. 한편, KM-402는 glucose에서는 가장 ethanol 생산성이 좋았으나 xylose 및 cellobiose에서는 생산하지 못하였다. There strains able to efficiently produce ethanol from cellulosic hydrolysates were isolated from soil samples by enrichment culture in liquid saccharified wheat bran medium. The profiles of physiological and biochemical properties of two yeasts KM-09 and KM-402 and a bacterium Hg-225 were almost identical from those of Candida sp. and Klebsiella sp., respectively. Strains KM-09 and HG-225 used xylose and cellobiose as fermentable sugars, and HG-225 had a wide range of sugar utilization for ethanol fermentation. The optimal pH and temperature for growth of KM-09, KM-402 and HG-225 were 5.8, 5.6 and 6.8 and 32℃, 30℃ and 38℃, respectively. During the ethanol fermentation in saccharified wheat bran by the isolated strains, optimal temperature for ethanol production was more or less higher than those for growth, and addition of 0.2%(w/v) MgSO_4 into the medium enhanced ethanol productivity. Of the three strains ethanol content of KM-09 was the highest with about 2.3%(v/v), and ethanol production rate of HG-225 was faster than the others and maximum productivity was after 4 days. KM-09 (1.42%, v/v) and HG-225 (1.05%, v/v) produced ethanol from 4%(w/v) xylose but growth rate was slower than on glucose. Otherwise KM-402 showed the highest ethanol productivity on glucose, but no ethanol was detected on xylose and cellobiose.

방선균 분리주가 생산하는 Phospholipase C 저해물질인 MT2617-2B의 분리 및 특성

고학룡,이현선,오원근,안순철,김보연,강대욱,민태익,안종석 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1

방선균 분리주 MT2617-2의 배양액으로 부터 phospholipase C (PLC) 저해물질인 MT2617-2B를 n-butanol 추출 및 column chromatography 법을 이용하여 분리하였다. MT2617-2B는 IR ^13C- 및 ^1H-NMR 그리고 ESI-MS에 의한 구조분석 결과, 한 개의 hemiketal ring, polyhydroxyl 및polymethyl groups으로 구성되었으며 side chain으로 한 개씩의 malonate 및 guanidine group을 가지는 분자량 1057의 macrolide 화합물이었다. 따라서, MT2617-2B를 기존의 macrolide 항생제인 copiamycin 및 niphithricin A로 동정하였다. 한편, MT2617-2B는 methanol 용액에서 실온에서 방치하였을 때 도일한 분자량을 가진 두 개의 이성질체를 생성하였다. PLC γ1과 -β1에 대해 각각 25 및 50㎍/㎖의 IC_50 값을 가지며, Staphylococcus aureus 와 Candida albicans에 대해서는 항균활성을 나타내지만 Escherichia coli에는 나타내지 않았다. A phospholipase C (PLC) inhibitor (MT2617-2B) was isolated from the culture broth of actionmycetes isolate MT2617-2 by the extraction with n-butanol and column chromatographic techniques. The molecular weight of the inhibitor was 1057, by the spectroscopic analyses of IR ^13C- and ^1H-NMR and ESI-MS. The chemical structure of MT2617-2B was found to be a macrolide compound consisted of a hemiketal ring, polyhydroxyl and polymethyl groups, which had a malonate and guanidine group as its side chain. MT2617-2B produced its two isomers having the same molecular weight by standing in methanol solution at room temperature. Therefore, MT2617-2B was identified as copiamycin and niphithricin A, macrolide antibiotics. The values of IC_50 against PLC γ1 and PLC-β1 were 25 and 50㎍/㎖, respectively. MT2617-2B had antimicrobial activities against Staphylococcus aureus and Candida albicans, but not against Escherichia coli.

Pseudomonas aeruginosa 3120 으로부터 항생물질의 생산 , 분리 및 특성

고학룡(Hack Ryong Ko),전효곤(Hyo Kon Chun),고영희(Yung Hee Kho),성낙계(Nack Kie Sung) 한국응용생명화학회 1993 Applied Biological Chemistry (Appl Biol Chem) Vol.36 No.6

A strain that inhibited the growth of Pellicularia sasakii was isolated from the soil and identified as Pseudomonas aeruginosa 3120. A dark brownish antibiotic. MRL3120 isolated and purified from the culture broth of P. aeruginosa 3120 was soluble in ethylacetate, chloroform and methanol, and it was active against gram-positive and negative bacteria as well as fungi. The structure of MRL3120 was identified as a chelate compound consisting of two N-methyl-N-thioformyl-hydroxylamine and a copper ion by the analysis of UV. IR, and EI-MS spectra and other physico-chemical properties and supposed to have a structure of fluopsin C related compound. Addition of CuSO₄ into the fermentation medium containing soybean meal increased antifungal activity but no activity was found in the presence of EDTA (0.1%, v/v). However antibiotic MRL3120 was not produced in the fermentation medium containing soytone instead of soybean meal but it was rapidly produced by the addition of CuSO₄.

식물정유, 길항세균 및 난황유를 이용한 시금치 노균병 방제

이정한,정성우,전성식,강신권,최준민,임채신,고학룡,이광수,정남준,배동원 경상대학교 농업생명과학연구원 2012 농업생명과학연구 Vol.46 No.2

본 연구는 친환경적으로 시금치 노균병을 방제하기 위하여 식물정유, 길항미생물 및 난황유의 병발생 억제효과를 검정하여 화학농약 대신에 이용할 수 있는 친환경제제로서의 가능성을 검토하였다. 5종의 식물정유 가운데 노균병 방제효과는 1회 처리구와 1주 간격 2회 처리구 모두 Eucalyptus approximans 오일의 병원균 억제효과가 60% 정도로 가장 높은 것으로 나타났다. 반면에 토양에서 선발된 길항균 세 종류의 시금치 노균병 방제효과는 모두 20% 이하로 식물정유 처리보다 낮게 나타났다. 난황유 0.3%와 0.5%농도에서 1주일간격 2회 처리구에서 병원균 억제율이 각각 84.5%와 91.5%로 억제되어 가장 효과가 우수한 것으로 나타났다. In this study, we evaluated environmental-friendly disease control methods using essential oils, antagonistic bacteria, or cooking oil and egg yolk mixture (COY). Among the five tested plant essential oils, Eucalyptus approximans oil shown the highest effect to control of the downy mildew disease (60% disease control value). Interestingly enough, well known antagonistic bacteria against various fungal pathogens were appeared lower control efficiency than plant essential oils. In field conditions, the cooking oil and yolk mixture (COY) treatments shown significantly decreased the downy mildew incidence with 0.3% (disease conttrol value 84%) and 0.5% concentration (disease control value 91%). Our findings suggest that the COY has a great potential as the eco-friendly downy mildew disease control formulation.

Pseudomonas Aeruginosa 가 생산하는 항생물질의 분리 및 특성

고영희,고학룡,전효곤,이호재,성낙계 한국미생물생명공학회 1993 한국미생물생명공학회 학술발표초록집 Vol.1993 No.1