慶北地域에서 栽培中인 芍藥의 Paeoniflorin 含量分析

이인구,주길재,김태강,정재동 慶北大學校農業科學技術硏究所 1996 慶北大農學誌 Vol.14 No.-

主재배품종인 의성종과 영천종의 유효성분을 HPLC 및 TLC로 분석한 결과 主성분인 paeoniflorin 함량은 의성종이 영천종보다 높으며 albiflorin과 oxypaeoniflorin의 함량을 두 재배종에서 비교하면 의성종은 albiflorin 함량이 많고 영천종은 oxypaeoniflorin의 함량이 많았다. 생산지에 따른 paeoniflorin의 함량차이는 2.15%에서 4.08%의 범위였고 의성과 거창의 것은 비슷하였으며 영천의 것이 가장 낮았다. 18종의 재배종들은 품종에 따라 다양한 paeoniflorin 함량 차이를 보이며 작약성분분석의 HPLC 크로마토그람을 albiflorin, oxypaeoniflorin 및 paeoniflorin피크를 기준으로 비교해본 결과 크게 3가지 패턴으로 분류할 수 있었다. 이 세가지 유형은 의성종 패턴 9종, 영천종 패턴 4종 풍기표준 종패턴 5종 이었다. 수확시기에 따른 작약의 paeoniflorin의 함량변화를 조사한 결과 3월과 9월에 수확한 것이 낮았으며 5월과 11월에 수확한 작약의 paeoniflorin 함량이 높았다. The effective components of the peony cultivar, Euseongjagyag and Youngcheonjagyag, which were the major cultivars of peony in Kyeongbuk area, were determined with HPLC and TLC. The paeoniflorin content in the root of Euseongjagyag were more than that of Youngcheonjagyag. The root of Euseongjagyag contained much albiflorin and Youngcheonjagyag contained much oxypaeoniflorin in comparision with albiflorin and oxypaeoniflorin contents in both cultivars of peony. Paeoniflorin contents in accordance with peony prodution regions were ranged from 2.15% to 4.0%, and paeoniflorin content of local cultivar of Euseong and that of Geochang were approximatly the same but that of Youngcheon was the lowest. Paeoniflorin content in the 18 accessions of peony cultivar which were collected from Kyeongbuk area and harvested on November 1993, were ranged from 1.41% to 5.30%. The 18 accessions of peony were classified with the HPLC chromatogram pattern of peony root extract into the three groups which composed with Euseong peony group(9 accessions), Youngcheon peony group (4 accessions) and Punggi standard peony group(5 accessions). High content of paeoniflorin was contained in peony root harvested in May and November, but low content of paeoniflorin was contained in peony root harvested in March and September.

혈슬관절 및 외상성 활액막염 환자에서 활액막의 변화

김인,이승구,김수근 최신의학사 1983 最新醫學 Vol.26 No.4

방선균 Streptomyces chibaensis J-59 Xylanase의 정제 및 자일로 올리고당(Xylooligo-Saccharides)의 생산

주길재,이인구 慶北大學校農業科學技術硏究所 1996 慶北大農學誌 Vol.14 No.-

Streptomyces chibaensis J-59 xylanase는 CSL 배지 (1% corn steep liquor, 0.15% glucose, 0.1% MgSO₄·7H₂O, pH 7.0)에서 1% xylan을 효소생산 유도물질로 첨가하였을 경우에 생산(0.83 unit/ml)되었으나, xylose를 비롯한 각 종 탄소원을 포함하는 배지에서는 효소가 생산되지 않았다. S. chibaensis로부터 생산된 세포외 xylanase를 (NH₄)₂SO₄분획침전, DEAE-Sephadex A-50 column chromatography, Sephadex G-200 gel filtration 과정으로 약 29%의 수율로 20배 정제하였다. 정제한 xylanase는 SDS-PAGE 및 Sephadex G-200 gel filtration에서 분자량이 모두 25,000 Da을 나타나 monomeric enzyme으로 확인되었다. 금속이온에 대한 영향은 ?? ??, ??, sodium dodecyl sulfate, N-bromosuccinide 등에서는 아주 강한 저해를 받았고, ??, ??에서는 약간의 저해를 받았다. 반면에 ??는 효소활성을 증가시켰다. Xylanase에 의한 xylooligo 당의 생산양상을 TLC로 조사한 결과 xylobiose, xylotriose 및 xylotetrose가 주생산물이었으며, 반응 1시간 이후부터 소량의 xylose 생성되었다. 따라서 본 효소는 전형적인 endotype xylanase로 확인되었으며 새로운 기능성식품인 자일로 올리고당(xylooligo-saccharides)의 제조에 이용할 수 있을 것이다. S. chibaensis J-59 produced an extracellular xylanase in a CSL medium composed of 1.5% corn steep liquor, 0.1% MgSO₄·7H₂O, 0,012% CoCl₂·6H₂O, and 0.15% glucose containing xylan. but it did not produce in the culture medium containing xylose. The production of enzyme reached to a maximum level (0.83 uints/㎖) when bacteria were cultured in 2.5ι jar fermentor for 48hrs at 30℃ and pH 7.0 Furthermore, S. chibaensis J-59 produced an intracellular glucose isomerase in a medium containing xylan and/or xylose. Xylanase was purified 29-fold over the culture supernatants of S. chibaensis J-59 by ammonium sulfate fractionation, chromatography on DEAE-Sephadex A-50, and gel filtration on Sephadex G-200. The purified enzyme is a monomeric enzyme with a native molecular mass of 25 kDa and a subunit molecular mass of 25 kDa. The purified enzyme requires ?? for activity ??, ??, is not an inhibitor but inhibit by ??, ??, and ??, sodium dodecyl sulfate, N-bromosuccinide. Pattern of hydrolysis demonstrated that the xylanase was an endo-splitting enzyme able to break down birchwood xylan at random giving xylobiose, xylotriose and xylotetrose as the main end products.

기능성 Zeolite-KCIO₃ 복합체에 의한 Chlorothalonil의 분해

최충렬,박만,이동훈,이병묵,이인구,최정,김장억 한국환경농학회 2004 한국환경농학회지 Vol.23 No.2

Salt occlusion in zeolite is a unique phenomenon that takes place only when the salt size is similar to the window size of host zeolite. KClO₃-occluded zeolite, as an environment-friendly oxidant, has a high potential for effective removal of various organic pollutants. This study was carried to investigate the characteristics and the removal kinetics of fungicide chlorothalonil by zeolite-KClO₃ complex. About 10% of KClO₃ was occluded in zeolite pores synthesized by salt-thermal method from fly ash, although the occlusion amount was relatively less compared to that of nitrate salts. By occlusion with KClO₃, no remarkable changes were found in X-ray diffraction patterns of cancrinite, whereas some decrease of overall peak intensities was found with those of sodalite. Different releasing kinetics of ClO^(-)₃ ion were observed in distilled water and soil solution from zeolite-KClO₃ complex. Two reactions, hydration and diffusion, seem to be related with the release of KClO₃. Therefore, the release isotherm of ClO^(-)₃ ion well fitted to the power function model which indicate the release was made by hydration and diffusion. The removal of chlorothalonil by zeolite and KClO₃ reached at reaction equilibrium within 6 hours by 18% and 47%, respectively. However, the chlorothalonil removal by the zeolite-KClO₃ complex increased slowly and steadily up to 92% in 96 hours. These findings suggested that zeolite-KClO₃ complex could be applied for effective removal of organic contaminants in the soil and aqueous environment.

Heterologous plasmid DNA 의 transformation에 있어서의 細胞膜 蛋白質의 기능에 對하여

徐正塤,洪淳德,李麟九,金英浩 慶北大學校 1983 論文集 Vol.36 No.-

Terramycin resistance plasmid DNA and Streptomycin resistance plasmid DNA have been prepared from Streptomyces rimosus and E. coli KPM 105, respectively, by phenol extraction of lysozyme-lysed cells. And the plasmid DNA was introduced into B. subtilis KPM 73, B. subtilis BD 224, B. subtilis 110, B. subtilis RM 125, B. amyloliquef aciens and B. megaterium IAM B425 by transformation. The variable characters affecting heterologous plasmid transformation, and the functions of competence factor and binding factor existing in the recipient cell surface were futhermore studied. Terramycin resistance plasmid was well transformed into Bacillus subtilis KPM 73 and B. subtilis RM 125, and Streptomycin resistance plasmid was well transfomed into B. subtilis BD 224, B. subtilis 110 and B. megaterium IAM B 425 at high frequency. The high frequency of plasmid transformation was obtained at 4 hr of incubationin growth medium. 20 to 50 min in competence medium, and the optimal pH and temperature for competence were 7.0 and 20C to 30C, respectively. The transformation of B. subtilis KPM 73 reached a maximum level after 20 min of exposure to DNA, B. subtilis BD 224 after 30min, and B. subtilis RM 125 after 10 min. Ca, Cu, Fe, Mg and Mn ions increased the transformation frequency of B. subtilis KPM 73 but Hg, Zn and Co ions decreased the transformation frequency. Ca ions were required for transformation of E. coli C600 and tranrsformation frequency was increased 20 times by addition of 75mM CaCl_2. The competence development of recipient cell was enhanced by addition of lysozyme in the competence medium and lysozyme was more affective in the early stage of competence development than the late stage of competence development, in which the spontaneous competence development reached maximum level. In sight of these observations, competence factor would seem to be a kind of cell wall lytic enzyme such as lysozyme. when the protoplasts of B. subtilis KPM 73 were transformed by terramycin resistance plasmid, the protoplasts were successfully transformed and the transformation frequency was 10^-2. The competent cells were treated with small amount of protein-digesting enzyme such as trypsin, and then exposed to transforming plasmid DNA. The recipient cells that were treated with trypsin were transformed 10 to 100 folds lower than non-trypsin-treated control cells. These results show that the binding factor must be a protein or at least a complex containing protein that is essential to it's activity. And external protein such as bovine albumin was effective on transformation of trypsin treated cell, suggesting that the cell surface-located binding factor did not have a specificity and the binding factor could be replaced by external substance such as bovine albumin. Terramycin resistance plasmid extracted from Ter^R transformant of B. subtilis KPM 73 was well transformed into B. subtilis KPM 73. And the electrophorsis pattern of Ter^R plasmid extracted from Ter^R transformant of B. subtilis KPM 73 was equal to that from St. rimosus.

Mesoporous Titanium Oxo-Phosphate에 의한 살균제 Chlorothalonil의 광분해

최충렬,김병하,이병묵,최정,이인구,김장억 한국환경농학회 2003 한국환경농학회지 Vol.22 No.4

Mesoporous titanium oxo-phosphate(Ti-MCM)은 기존의 유기성 오염물질의 광분해제로 널리 이용되고 있는 TiO₂에 비해 표면적이 매우 넓은 장점이 있다. 그러므로 본 연구에서는 Ti-MCM에 의한 chlorothalonil의 흡착 및 광분해 특성을 TiO₂와 비교하였다. 합성된 Ti-MCM은 hexagonal 형태로 d-spacing이 4.1 ㎚이었다. 암조건에서 TiO₂에 의한 chlorothalonil의 흡착은 거의 일어나지 않았으나, Ti-MCM에 의한 흡착은 반응 1시간까지 28%로 급격히 증가하여 흡착평형에 도달하였다. UV조사 하에서 반응 9시간 후의 TiO₂와 Ti-MCM에 의한 chlorothalonil의 제거율은 각각 88%와 100%로 나타났다. 그러나 정치상태에서의 광분해 속도는 chlorothalonil과 Ti-MCM사이의 낮은 접촉에 의한 반응성의 감소로 느린 경향을 나타내었다. 또한 Ti-MCM에 의한 chlorothalonil의 분해효율은 용액의 초기 농도가 낮을수록, pH 7까지 반응용액의 pH가 높을수록 증가하였다. Titanium mesoporous materials have received increasing attention as a new photocatalyst in the field for photocatalytic degradation of organic compounds, The photocatalytic degradation of chlorothlonil by mesoporous titanium oxt-phosphate (Ti-MCM) was investigated in aqueous suspension for comparison with TiO₂ (Degussa, P25) using as an effective photocatalyst of organic pollutants. Mesoporous form of titanium phosphate has been prepared by reaction of sulfuric acid and titanium isopropoxide in the presence of n-hexadecy(trimethylammonium bromide. The XRD patterns of Ti-MCM are hexagonal phases with d-spacings of 4.1 nm. Its adsorption isotherm for chlorothalonil reached at reaction equilibrium within 60 min under dark condition with 28% degradation efficiency. The degradation ratio of chlorothalonil after 9 hours under the UV radiation condition (254 nm) exhibited 100% by TI-MCM and 88% by TiO₂. However, these degradation kinetics in static state showed a slow tendency compared to that of stirred state because of a low contact between titanium matrices and chlorothalonil. Also, degradation efficiency of chlorothalonil was increased with decreasing initial concentration and with increasing pH of solution. As results of this study, it was clear that mesoporous titanium oxo-phosphate with high surface area and crystallinity could be used to photocatalytic degradation of various organic pollutants.

고추 근권에서 분리한 Bacillus subtilis J-24의 검은 곰팡이병원균 Alternaria alternata에 대한 길항력 및 고추의 초기 생육에 미치는 영향

주길재,김학윤,허상선,우철주,이인구 경북대학교 농업과학기술연구소 1999 慶北大農學誌 Vol.17 No.-

Thirty bacteria were isolated from the red pepper rhizosphere. The isolates were screened for antagonism to Alternaria alteranta causing red pepper black rot. Antagonistic bacterium No. J-24 was selected among the isolated bacteria and was identified as Bacillus subtilis based on morphological and physiological characterisitics and MIDI system. B. subtilis J-24 showed antifungal activities against A.alternata(inhibition percentage, 99%), Botrytis cinerea, Phytophthora capsici, Pythium ultimum, Colletotrichum gloeosporioides, Stemphylium botrysum. The growth of red pepper seedling was promoted as compared to control when the microbial inoculants was mixed in bed soil. In the mixed microbial inoculants bed soil, the leaf area of red pepper was increased of 15 percent, the hypocotyl weight 12 percent, the root length 12 percent, total dry weight 13 percent as compared to those grown in the genenal bed soil.

Purification and Properties of β-Galactosidase Produced by Gongronella sp.

李麟九 대구효성가톨릭대학교 1982 연구논문집 Vol.25 No.1

土壤으로부터 分離한 곰팡이인 Gongronella sp. L79-1이 生産하는 β-galactosidase를 DEAE-cellulose ,DEAE-Sephadex A-50 및 Sephadex G-100 column chromatography로 純粹하게 精製하였다. 精製된 酵素를 Sephadex G-200으로 젤 여과한 結果 그 分子量이 110,000이었다. o-nitorphenyl-β-D-gal-actopyranoside(ONPG)를 基質로 본 酵素의 最適 pH를 測定한 結果 3.0~4.2였으며, 乳糖을 基質로 했을 경우 最適作用 pH는 3.6~4.8이었다. 本 酵素는 pH1.4~6.4사이에서 40℃ 3時間 處理에도 安定하였다. 酵素作用 最適 溫度는 55℃附近이었으며, 活性化에너지가 8kcal/mole이었다. 本 酵素는 50℃에서 1時間 處理시켜도 전혀 失活이 없었으나 60℃에서는 10分만에 活性의 1/2이 破壞되었다. 本 酵素의 作用이 Mn²+ 에 의해 촉진되고, Hg²+ 과 sodium dodecylsulfate에 의해 沮害되었다. ONPG, p-nitrophenyl-β-D-galactopyranoside 및 乳糖의 Km는 各各 0.8, 0.7, 8.1mM이었으며 本 酵素는 α-D-galactoside에는 전혀 作用하지 못했다. 本 酵素는 glucose에 의해서는 沮害를 받지 않으나 galactose에 의해서는 混合型의 沮害作用을 받았다. 本 β-galactosidase는 2% 乳糖을 72%까지 分離할 수 있었다. Extracellualr β-galactosidase was produced on the liguid medium by Gongronella sp. L79-1, which was isolated from soil. The β-galactosidase was purifoed to be chromatographically homogeneous by DEAE-cellulose, DEAE-Sephadex A-50, and Sephadex G-100 column chromatography. Molecular weight of the enzyme was determinded to be 110,000 by the gel filtration of Sephadex G-200. The optimal pH for the enzyme reaction was 3.0 to 4.2 with o-nitorphenyl-β-D-gal-actopyranoside(ONPG) as a substrate and 3.6 to 4.8 with lactose as a substrate. The enzyme was stable at pH ranges between 1.4 and 6.4 at 40℃ for 3hr. The optimal temperature for the hydrolysis of ONPG by β-galactosidase was 55℃. The activation energy was 8kcal/mole. The enzyme was stable at 50℃ for 1 hr and denatured lograithmically wiht a half decay of 10 min at 60℃. The enzyme reaction was activated by Mn²+ and inhibited by Hg²+ and sodium dodecylsulfate. The Km values were 0.8, 0.7 and 8.1 for ONPG, p-nitrophenyl-β-D-galactopyranoside and lactose, respectively. The enzyme was not active to α-D-galactoside. The enzyme was inhibited by galactose as a mixed competitive-noncompetitive type, but not by glucose.

Cyclohexanone 資化菌의 分離 및 그 特性

李麟九 대구효성가톨릭대학교 1981 연구논문집 Vol.23 No.3

A bacterium which grows on cyclohexanone as sole carbon and energy source was isolated from sludge of industrial areas in Daegu and identified as Acinetobacter calcoaceticus C-10. The growth medium for the optimal culture condition was composed of 0.5% cyclohexanone, 0.33% (NH_4)_2HPO_4, 0.05% KH_2PO_4, 0.2% K_2HPO_4, 0.02% MgSO_4ㆍ7H_2O, and 0.05% yeast extracts. The culture was incubated at 30℃ for 24 to 48 hr on the shaker. The optimal pH value for the growth was 7.2 to 8.0. the bacteria utilizedethanol, 1-butanol, 1-pentanol, 1-hexanol, and cyclohexanol as a carbon source but not methanol, m-cresol, glycerol, and cyclohexane. the bacteria grew on acetate, citrate, and adipate but not on salicylate and phthalate. All kind of sugars except xylose were nt catabolized. Whole cells of Acinetobacter calcoaceticus C-10 grown on cyclohexanone oxidized cyclohexanone, cyclohexan-1,2-dione, cychohexnol, cyclohexan-1,2-diol, and ε-caprolactone with high Qo_2 values, bbut not cyclohexene oxide, cyclohexane monocarboxylic acid, and cyclohexan-1,2-dicarboxylic acid. Extracts of cyclohexanone-grown cells contained NAD-linked cyclohexanol dehydrogenase which catalized the oxidation the cyclohexanol to cyclohexanone.

優良室素固定菌의 探索分離 및 生理的 特性

朴愚喆,李光熙,金進鎬,李麟九,曺晋基 慶北大學校出版部 1987 慶北大農學誌 Vol.5 No.-

This experiment was conducted to search for Rhizobia with good nitrogen fixation abilities and to investigate their physiological characteristics isolated from 12 soybean cultivars and the affinities of root nodule bacteria with soybeans. The results obtained were as follows ; Based on colors, Rhizobia grown on YMA medium were divided into 3 groups, i. e., white, translucent and transparent, amounting to 60, 30 and 10%, respectively. In litmus milk reation, the strains which produced alkali, acid serum, alkaline serum and acid reached to 51, 29, 9 and 11%, respectively. Strains, S022, and S096 were slow-growers and produced alkaili, while strains, S080, S090, and S118 were fastgrowers and produced acid. The growth of root nodule bacteria on YMA medium was favorable between the initial pHs of 6.0~7.0. Glutamine, asparagine and allantoin as nitrogen sources enhanced the growth of root nodule bacteria. All the strains tested formed nodules on the soybean roots, and the strains with good symbiotic nitrogen fixation abilities that had white color, small colony, nitrate reduction abilities and no nitrite reduction abilities showed comparatively high nitrogen fixing activities. Some strains varied in nitrogen fixing activities according to soybean cultivars, and a few strains formed ineffective nodules which showed no nitrogen fixing activity.