A Study on Dextranse from Aspergillus sp.BY-54 Strain : I. Cultural Conditions and Properties

방병호 서울保健大學 1993 論文集 Vol.13 No.1

방사선 BY-445가 생성하는 α-amylase 저해물질의 특성

방병호,이진영 서울보건대학 1997 서울보건대학 부설 한국보건과학연구소 논문집 Vol.4 No.1

α-amylase에 대한 강력한 저해물질을 생산하는 방사균 BY-445를 토양으로부터 분리하였으며 이 균이 생산하는 배양액을 Amberlite IRA 420 column, Silica gel column chromatography 및 Sephadex G-50 column을 이용하여 조정제하였다. 이때 수율은 21.4%였으며 비활성도는 6배 증가하였다. 그리고 저해제의 α-amylase 100㎍에 대해 2배의 농도인 200㎍에서 저해활성이 거의 95%로 최고에 달하였다. 본 저해물질의 활성에 미치는 금속이온의 영향으로 Co^(+2), Hg^(+2), Pb^(+2), 및 Cu^(+2) 등에 의해 상대활성이 각각 13, 30, 37 및 72%로 각각 나타났다. 또한 본 저해물질은 열에 대한 안정성이 커서 100℃에서 120분간의 열처리를 하여도 그 활성은 거의 잔존하였으며 또한 전 pH 범위(pH2.0~12.0)에서도 아주 안정하였다. 본 저해물질의 여러가지 생화학적 반응을 검토한 결과 Anthrone, Molisch, Phenol-sulfuric acid반응에서는 양성을 띠었으며, Ninhydrin, Biuret, Benedict 등의 반응에서는 음성으로 나타났으며, 각종 carbohydrases에 대한 본 저해물질의 활성을 검토한 결과 사람 침 α-amylase와 Bacillus subtilis,porcine pancreas,Asp.niger의 glucoamylase등의 α-amylase에 대한 저해율이 각각 87%와 81,79,63%로 나타났으며, Aspergillus속의 cellulase와 Saccharomyces cerevisiae의 invertase에 대한 저해는 나타나지 않았다. A strain of Actinomycetes BY-445 isolated from soil was able to produce a biological active substance that has a strong inhibitory activity against hydrolysis by α-amylase. Extracellular inhibitory substance (inhibitor) from the culture broth of Actinomycetes BY-445 was purified to partial homogeneity by procedures including Amberlite IRA 420 column, Silica gel column, and Sephadex G-50 column chromatography. The partial purifying procedures resulted in 6-fold purification with the overall yield of 21.4% and the velocity of the inhibitor was reached at maximum in inhibitor concentration of 200㎍ against α-amylase enzyme concentration of 100㎍. In effect of metal salt, Co^(+2), Hg^(+2), Pb^(+2) and Cu^(+2), the relative inhibitory activity were shown about 13, 30, 37 and 72%, respectively and the inhibitor was also appeared to be relatively thermostable, and no appreciable inactivation was observed after incubation at 100℃ for 2 hour. The substance was stable in pH range from 2.0 to 12.0 at 37℃. The biochemical reaction of the substance was positive to Anthrone, Molisch and Phenol-sulfuric acid, but negative to Ninhydrin, Biuret and Benedict. A strong inhibitory activity against hydrolysis by α-amylase of human saliva, Bacillus subtilis, and porcine pancreas and glucoamylase of Aspergillus niger was shown 87, 81, 79 and 63%, respectively. but it did not inhibit cellulase of Aspergillus sp. and invertase of Saccaromyces cerevisiae.

Properties of Glucose Isomerase from Arthrobacter sp. L-3

방병호,남궁석,이정윤,윤은숙,이상건,이은숙 서울保健大學 1991 論文集 Vol.11 No.1

Physiological Properties of Thialysine Resistant Mutant of Candida utilis (V)

방병호 서울保健大學 1985 論文集 Vol.5 No.1

Thialysine 내성변이주(TRN-4006)를 야생주(NCYC-359)로 부터 N-methyl-N'-nitro-N-nitrosoguanidine(NTG)처리하여 얻었다. 이 변이주에 의한 lysine 흡수속도가 야생주에 비해 훨씬 빨랐으며, lysine흡수에 영향을 주는 여러가지 요소를 검토하였다. lysine흡수는 glucose와 같은 에너지 자원이 공존시 훨씬 증가되었고 산처리 후 lysine 흡수는 무처리구에 비해 감소되었으며 또한 lysine 흡수가 암모니아에 의해 저해되었다. 기본배기에 lysine을 첨가한 후 배양시 무첨가구에 비해 lysine 흡수속도가 크게 줄어들었다. 34%의 sucrose로 질소원의 고갈상태로 된 세포의 lysine 흡수속도는 현저히 증가되었다. Thialysine resistant mutant (TRN-4006) was induced from the wild strain of Candida utilis (NCYC-359) by N-methyl-N-nitro-N-nitrosoguanidine (NTC) treatment. The velocity of lysine uptake by this mutant was increased as compared with wild type strain. The effect of various factors on lysine uptake was investigated. Lysine uptake was increased in presence of an energy source such as glucose. After acid treatment, lysine uptake decreased as compared with no treatment. The rate of lysine transport was reduced by ammonia. Lysine uptake by cells preloaded with lysine was decreased significantly. The intracellular lysine pool was depleted to nondetectable levels, usually within 5hr, when the cells were exposed to 34% sucrose. Lysine uptake by these starved cells also increased.

Aspergillus species (B-17)균주가 생산하는 Inulin 분해 효소에 관한 연구 : 제1보. 고체배양에 따른 배양조건

방병호,윤치영 서울保健大學 1986 論文集 Vol.6 No.1

Properties of Inulase from Aapergillus sp.B-17

방병호,윤치영 서울保健大學 1988 論文集 Vol.8 No.1

방선균 BY-019에 의한 광범위 항생물질의 생산에 관한 연구 : 최적 생산 조건

방병호 凡石學術奬學財團 1997 凡石學術論文集 Vol.1 No.1

광범위 항생제를 개발하기 위하여 토양으로부터 600주의 방선균을 분리하였으며 이들 배양액으로부터 E. coli. Bacillus subtilis 및 Candida albicans를 사멸시킬 수 있는 항생물질를 생산하는 방선균 BY-019 균주를 선별하였다. 이 균으로부터 항생물질 생산 최적 조건을 검토하였으며 그 결과는 다음과 같다. 탄소원으로서 soluble starch가 가장 양호하였으며 다음으로 glucose, maltose, fructose, 및 sucrose 순으로 나타났다. 질소원으로서는 유기 질소원인 soybean meal이 가장 좋았다. 그러나 본 방선균 BY-019 는 무기질소원에서는 전혀 생육이 불가능하였다. Stearic acid, oxalic acid 와 같은 유기산의 첨가가 본 항생물질의 생산증가에 영향을 미쳤으며 금속염으로는 Mg^(2+) 이온이 항생물질 생산을 촉진시켰으며 Hg^(2+), Cu^(2+) 및 Zn^(2+) 등은 항생물질 생산에 대한 강한 저해제로 작용하였다. 또한 본 균의 항생물질 생산에 최적 pH와 온도는 각각 pH 7.0과 35℃에서 3일간의 진탕배양에서 항생 물질 생산이 최고에 달하였다. In effect to develop effective broad spectrum antimicrobes, I screened over 600 isolates of Actinomycetes from soil, and selected BY -019 as strain producing broad spectrum substances against E. coli. Bacillus subtilis and Candida albicans. I investigated the optimal culture conditions for production of the broad spectrum antibiotics from BY-019 strain. The results are as follow: It was found that 2% soluble starch was good as carbon source, and glucose, maltose, fructose and sucrose can also be utilized as the carbon sources although their availabilities are less than that of soluble starch in this order. As nitrogen source, it was shown that soybean meal. organic nitrogen source was the most suitable, but BY-strain was not grown on inorganic nitrogen source at all. Stearic acid and oxalic acid had slight effect in production of antibiotic. The production of antibiotic was also activated by Mg^(2+) ion but inhibited by Hg^(2+), Cu^(2+) and Zn^(2+). The optimum initial pH and temperature on production of antibiotic were pH 7.0. 35℃, respectively.

울진원자력발전소 주변 소나무림의 종조성과 군락 특성

방제용,전영문,변두원,류병혁,이호준 建國大學校基礎科學硏究所 1995 理學論集 Vol.20 No.-

울진원자력발전소 주변 소나무림의 종조성과 군락특성을 1994년 3월부터 95년 2월까지 조사한 결과는 다음과 같다. 소나무군락은 상수리나무하위군락과 산철쭉하위군락으로 분류되었으며, 전조사방형구 내에서 출현한 종수는 총 116종으로 상수리나무하위군락과 산철쭉하위군락에서 방형구당 출현종수는 각각 28종, 23종으로 나타났다. 소나무군락의 계층별 평균 수고와 식피율은 상수리나무하위군락에서 교목층이 각각 13.5m와 78.8%, 아교목층이 6m와 37.5%, 관목층이 1.9m와 53.6%, 초본층이 0.4m와 64.4%, 산철쭉하위군락에서는 교목층이 13m와 83.8%, 아교목층이 5.5m와 52.5%, 관목층이 1.9m와 57.5%, 초본층이 0.6m와 69.4%로 나타났다. 피복지수에 의한 계층별 우점순위는 교목층과 아교목층에서 각각 소나무가 8437.5, 1111.3으로 나타났고 관목층에서 진달래(2638.5), 초본층에서 큰기름새(1048.2)가 가장 높게 나타났다. 소나무의 DBH별 분포를 보면 11-15cm급이 상수리나무하위군락과 산철쭉하위군락에서 각각 29.1%(4.2 individuals/a), 26.7%(4.5 individual/a)로 가장 높게 나타났다. 현존식생도에 따른 각 식생형의 분포면적은 소나무림이 133.1k㎡(80.5%), 경작지와 주거지 28.7k㎡(16.2%), 곰솔림 1.8k㎡(1.1%) 순으로 나타났다. 녹지자연도의 등급별 분포는 7등급(이차림)이 70.4%, 2등급(경작지와 주거지) 17.4%, 8등급(이차림) 11.6%, 4등급(벌목지) 0.2%, 6등급(인공조림지) 0.1% 순으로 나타났다. Montreal model에 의한 소나무림의 식물현존량은 3,077,327.8ton으로 산정되었다. The floristic composition and community characteristics of Pinus densiflora forest in vicinity of Ulchin Atomic Power Plant were surveyed from March, 1994 through February, 1995. The results obtained are summarized as follows: The Pinus densiflora community was classified into the subcommunity of Quercus acutissima and Rhododendron yedoence var. poukhanense. The total number of species appearing in the entire surveyed quadrats was 116 sepcies, then 28 and 23 species per quadrat in Quercus acutissima subcommunity and Rhododendron yedoence var. poukhanense subcommunity, respectively. Tree height and vegetation cover by the layer of the Pinus densiflora community were 5m and 78.8% in tree layer, in subtree layer(6m, 37.5%), in shrub layer(1.9m, 53.8%) and in herb layer(0.4m, 64.4%) in Quercus acutissima subcommunity, and in tree layer(13m, 83.8%), in subtree layer(5.5m, 52.5%), in shrub layer(1.9m, 57.5%) and in herb layer(0.6m, 69.4%) in Rhododendron yedoence var. poukhanense subcommunity. The dominance sequences on each stratum determined by the coverage index showed that Pinus densiflora was most dominant in tree layer with 8437.5 and in subtree layer with 1111.3, Rhododendron yedoence var. poukhanense in shrub layer with 2638.5, and Spodiopogon sibiricus in herb layer with 1048.2 value. The highest value of diameter breast height(DBH) distritued in the range of 11~15cm in Quercus acutissima subcommunity(29.1%, 4.2 individuals/a) and Rhododendron yedoence var. poukhanense subcommunity(26.7%, 4.5 individuals/a), respectively. The distribution area of each vegetation type in the actual vegetation map was 80.5% in Pinus densiflora forest, 16.2% in cultivated land and residential section, and 1.1% in Pinus thunbergii forest. The distribution of the degree of green naturality(DGN) was 70.4% in the 7th grade(second forest), 17.4% in the 2nd grade(cultivated land and residential section), 11.6% in the 7th grade(second forest), 0.2% in the 4th grade(cutted area), and 0.1% in the 6th grade(afforestation). It was estimated that phytomass of Pinus densiflora in the study sites, which was calculated by Montreal model, was 3,077,327.8ton.

α-아밀라제 저해물질을 생성하는 방선균 BY-445의 동정

방병호 한국식품영양학회 1998 韓國食品營養學會誌 Vol.11 No.5

α-아밀라제의 강력한 저해물질을 생성하는 분리주 BY-445를 토양으로부터 분리하여 이를 동정하였다. 이 균주의 기균사는 나선형이며, 포자의 표면은 침상형(spiny)이었다. 멜라닌 색소 및 용해성 색소는 음성이었으며, 단백질 및 전분 분해력은 강한 편이었다. 세포벽 성분중 diaminopimelic acid는 LL형이었고 세포의 지방산은 16 : 0 iso가 25.30%, 15 : 0 anteiso는 16.19% 그리고 16 : 0이 13.16%였으며 이 균주는 Streptomyces violaceusniger와 유사하였으나 몇몇 배양학적, 생리학적 특성이 달라 Streptomyces sp. BY-445로 명명하였다. The strain BY-445 which produces new inhihitors of α-amylase was isolated from a soil sample and identified. The aerial hyphae of this strain develope in the form of open spirals. The spore chain of BY-445 strain appears in a spiral shape with spiny surface. Melanoid and soluble pigments were not observed. Gelatin was liquefied, and skim milk and starch was also hydrolyzed. The isolate contained LL-diaminopimelic acid in its cell wall hydrolysate. The content of fatty acid 16:iso, 15:0 anteiso and 16:0 was 25.30, 16.19 and 13.16%, respectively. BY-445 strain was closely related to Streptomyces violaceusniger but it was different from this strain in some cultural and physiological characteristics. This strain was, therefore, designated as Streptomyces sp. BY 445.

Trimeresurus flavoviridis Venom에 대한 Inhibitor검색

방병호 서울保健大學 1984 論文集 Vol.4 No.1

Trimeresurus flavoviridis 독에 대해서 저해물질을 생성하는 균 68-B주를 분리했으며 이 균주가 생성하는 물질이 조정제의 상대에서 사독에 대한 저해작용 및 성질에 대해서 검토한바 Trimeresurus flavoviridis독에 대한 저해율이 80~85%이었다. 본 시료가 열이냐 pH에 대해서 아주 인정하였으며 또한 기질의 종류에 따라 저해율에는 현저한 차이가 있었다. α-Chymotrypsin이나 pepsin에는 저해를 나타내지 않았으나 trypsin에서는 약간의 저해작용을 나타내었다. Cu^(++)나 Zn^(++) 이온이 반응액중에 공존시 그 저해능이 현저히 약화되었다. A substance which inhibit to the activity of Trimeresurus flavoviridis Venom protease was isolated from the cultured extract of mold. The active substance was crystalized with the ethyl ether soluble fraction. About 85 percent of the protease activity was inhibited by this substance on hydrolyzing action of casein. This inhibitory substance was very stable to heat. For example, about 5 percent of the activity was decreased with treatment at 100℃ for 120minutes. This substance was acidic in solution but it was very stable in neutral or alkaline pH. The inhibitory activity was decreased by Cu^(+-+) and Zn^(++) on egg albumin and hemoglobin was 77 and 95 percent respectively. α-Chymotrypsin and pepsin were not inhibited by this substance, whereas trypsin was some what inhibited.