양파(Allium cepa)로부터 β-fructofuranosidase의 부분정제 및 특성분석

이용억 동국대학교 경주대학 1997 東國論集 Vol.16 No.1

양파의 구근으로부터 과당중합체(fructan)을 분해하는 효소를 ammonium sulfate 침전법, 이온교환 크로마토그래피, Con-A 흡착 크로마토그래피 그리고 gel-filtration 크로마토그래피 방법을 사용하여 부분 정제하였다. 이 효소는 inulin보다 sucrose를 빨리 가수분해하였으며 β-fructofuranosidase (invertase)로 확인되었다. 효소의 활성을 위한 최적 pH 및 온도는 각각 pH 5.5와 35℃ 이었다. 이 호산성 invertase는 lectin인 concanavalin A에 흡착하는 특성에 근거하여 당단백질인 것으로 사료된다. 이 효소는 열에 불안정하여 50℃에서 1시간 배양하면 활성을 상실하였다. 양파의 invertase는 ZnCl_2, HgCl_2 그리고 CuSO_4에 의해 부분적으로 활성이 억제되었다. A fructan-degrading enzyme was partially purified from onion (Allium cepa) bulbs by a combination of ammonium sulfate precipitation, ion-exchange chromatography, concanavalin-A affinity chromatography and gel-filtration chromatography. The enzyme hydrolyzed sucrose faster than inulin and was identified as a β-fructofuranosidase (invertase). The optimum pH and temperature for enzyme activities were pH 5.5 and 35℃, respectively. The acid invertase turns out to be a glycoprotein as evidenced by its ability to bind concanavalin-A (Con-A). The enzyme was heatlabile, with most activity being lost at 50℃ in 1 hr of incubation. The onion invertase was partially inhibited by ZnCl_2, HgCl_2 and CuSO_4.

Zea Mays L. 의 特殊成分과 澱粉의 X線 回析度

李容億,曺貞淳,申東旿 明知大學校 自然科學硏究所 1984 자연과학논문집 Vol.2 No.-

韓國産 메옥수수, 찰옥수수, 爆裂種에 대한 一般成分,無機質을 定量分析하고 또 胚芽油脂를 油出定量하고 그油脂의 物理,化學的 性質을 判斷하였다. 한편 各各의 녹말은 生 및 糊化시켜서 그의 X-線 회절도를 作成 比較 檢討하였다. 1.一般成分은 各各 Table 1에 表示한 바와 같다. 2.無機質成分은 各各 Table Ⅲ에 表示한 바와 같다. 3. 油脂의 物理化學的 性質 가.油脂의 色度差는 Table V에 表示한 바와 같다. 나.其他 粗脂肪, 比重, 굴절율, 요오드값, 산값, 검화값, 과산화물값, 불검화물값등은 Table Ⅳ에 表示한 바와 같다. 다.녹말에 대한 X-線 회절도의 試驗結果는 Table Ⅶ에 表示된 바와 같이 各 種間의 差異가 없었다. The general constituents and the mineral contents of White Dent Corn, Waxy Corn and Corn, all harvested in Korea, were first examined, and then the quality of each corn was examined through the test for the Physiochemical property of fat extracted from the germ of each corn. The result of qualitative comparison of starch, made by means of X-Ray Powder diffraction for the raw and the boiled starch of each corn, especially to find out any differences among the White Dent, the Waxy and the Pop Corn based on the point of starch, has reached the following conclusions; (1) The general consitutents and each content of the White Dent Corn are ; protein 10.3%, fiber 1.89%, Carbohydrate 69.93%, starch 64.5%, fat 5.36% and ash 1.65%. From it, it could be pointe out tthat the White Dent Corn contained more protein and ash than the others. Compared with the others, th Waxy Corn contained more Carbohydrate , showing its constitutents and contents as follows; protein 9.74f%, fiber 2.08%, Carbohydrate 71.04%, starch 65.5% fat 4.87% and ash 1.64%. As to the Pop Corn, however, it showed a relatively higher content of fat than the rest. Its constituents and each content were as follows ;protin 9.67%, fiber 2.19%, Carbohy-drate 69.39%, starch 64.0%, fat 6.62% and ash 1.64%. (2)Mineral Constituents; Regarding the mineral constiruents contained by the three kinds of corn which were sampled here for this study, the following results were noted; In case of the White Dent Corn, the content of P and Si were high, but the contents of Al and Zn were low. As for the Waxy Corn, the contents of Cu and Si were found less but the content of Al was found a little . Regarding the Pop Corn, however, the contents of its mineral constituents were higher in every respect than the other kinds of corn. Among them, especially , the contents of Si, Cu, Al, and Zn were a little higher. (3)Physiochemical Quality of Fat: The fat sampled here for the test was good enough with the one extracted through the method of pressure, oil pressing method, without any special refining process of high level.It was noted that the specific gravity of fat was 0.911 in the White Dent Corn, and 0.909 in the Pop Corn, which showed approximately 0.003 lower than the standard value specified b6 the Ministry of Health and Social Affairs. In the Waxy Corn, however, its value was almost as much as the specified level. As to the ditribution status of fat constituent, the Pop Corn contained the largest portion of all, showing that the Pop Corn 6.62%, the White Dent Corn 5.36% and the Waxy Corn 4.87% respectively. The iodine value of the sampled fat was 126 in the Waxy Corn, which belonged to the highest one of three kinds of Corn. And acid value of all the corn was generally a little higher than the standard required by the Ministry of Health and Social Affairs, since the fat was extracted from germ by means of ether and then was dried in the temperature of 90℃. In this case it was also noted that the Waxy Corn contained as much as the standard level. The White Dent Corn and the Pop Corn, however, wasd not up to the standard value. (4) Result of test for Starch X-Ray Power Diffraction: Starch, as its crystalline structure differs according to its kind, can be classified into three patterns such as A, B, and C. The raw starch (β-starch) sampled here belonged to A pattern and all the bolied starch (α-starch) to B pattern. When all the above were compared, all the sampled corn, White Dent, Waxy and Pop, did not show much difference in themselves.

고사리의 Shikimic acid 分析과 動物實驗(2) : Analysis of shikimic acid from bracken fern and the effect of bracken fern to mice

李容億,曺貞淳 명지대학교 1983 明大論文集 Vol.14 No.-

Study on the poisonous component of bracken fern. The investigator carried out the experiments for the purpose of finding out the effect of the bracken fern(BF) to mice. In this paper results about histological, nutritional and microbial researches of the bracken fern are reported. All experiments were carried out with bracken fern from Jae judo in 1979. First, those substances which were dried after boiling for 20 min dipping in water for 21hr. and extracted them from the water. After that, they were analyzed by AOAC method and spectrography method. The results showed that the backen fern of Jaejudo contains more fat and less protein, compared with that of Caernarvonshire and Sooweon. In respect to the minerals, K and Na are contained at a large amount and Si, Ca, Al, Fe, Ti, As and Mg at a relatively small amount. Evans, I.A. and Osman, M.A. in 1974 reported that the substance which caused carcinogenic effect was organic substance. The above team especially carried out the experiments to find out shikimic acid(3α, 4α, 5β-trihydroxy-1-cyclohexene-1-carboxylic acid). The edible portion of bracken fern was analyzed to find out shikimic acid by following methods. Sample A is drying fresh bracken fern, sample B is drying after boiling for 20 min and dipping in water for 21 hrs and sample C is drying after boiling for 10 min. Sample A showed 0.18 wt% of shikimic acid and sample of B,C showed value lessthan 0.001 wt% by the metohd of gas-liquid chromatography. To know the effect of shikimic acid to enteric bacteria, feces from each mouse weretested. So their enumerated bacteria were identified by API 20E kit and compared with the control mouse feces. All these identifications were carried out by means of selective media and API 20E kit. The identified bacteria were Escherichia coli, Enterobacter agglomerance, Enterobacter cloacoe, Citrobacter diverus-Levinea and Serratia rubidaea. Escherichia coli was dominant in control mice but it was reduces greatly in the treated mice. And in comparision with extracted components, pure shikimic acid showed the inhibitory effect on Escherichia coli. To find the effect when bracken fern were fed to mice, total 40 mice were divided into eight groups and feeding methods were pouring of BF powder and standard diet. In the former method, there was an increase in body weight of mice, but in the later method, not as former do. As a result of taking much fern by infusion method caused the result of not only decrease of weight but also much change around their months. That is, there was a phenomena like loss of bristle and the reduction of skin fur. In spite of the reduction of appetite, there was no change in the properties like degree of hardness, colour, smallness of feces in comparision with those of control. After above experiment, the histological changes were searched after six months feeding, the paraffin section of the tissues around around mouth and intestine were mainly stained by hematoxylen-eosin. The change of the skin tissues were the infiltration and the increase of the squamous-cells. The disapperance of the hair sheath was caused by the necrosis of the hair root, the infiltration of its cell, expention and hyperaemia of the blood vessels. And the symptoms like the edema and the tissue infiltration were developed in the hair root around the mouth. The increase of the giant cells in the spleen was rapid in contrast to controls and the differention of the nucleus was high. In the tissue of lungs, there was increase of lymphoid cells around the blood vessel and the small bronchus. Cell infiltrations were observed in blood vessel and glomerulus area in kidney. Noticeable facts were the enlargement of blood capilary and the hyperaemia of the tissues in lung, kidney, liver and stomach. It was generally accepted that the development of cancer like edema and carcinoma appeared in long term experiment i.e. at least six months, so the results of this experiment could be understood as an early symptom of above developments. And it might be thought as an important result that in skin tissue there were infiltration of squamous-cell and regressive changes around their mouths Consequently the appearance of phenomena like alopecia of elastic hair, skin and development of alopecia spots assumed to be the important results.

韓國産 高酸價 米糠油의 脫色에 關한 硏究

李容億,金恩卿,盧長淑 명지대학교 1983 明大論文集 Vol.14 No.-

The rice bran produced in the process of rice milling is only ingredient of edible oil and fat in Korea. If and when the rice bran is left negligently for a long time, a good quality of edible oil cannot be obtained. It is not only because the loss of the rice bran is resulted in the process of refining from starch degeneration, discoloration, and generation of free fatty acid or unsaponifiable matter but because the decolorization processing becomes much more difficult. In this connection, the experiment on the decolorization of the high acid value rice bran oil was made with clay. Although remarkable results were not yielded, continuous research is needed. If successful, it will contribute to the development of domestic medical ingredients with consequent extraction and refining of orizanol or inositol etc.

신규 살균제 KNF 1002의 오이 및 고추 중 잔류특성

김태화,이재영,유용만,김장억 한국환경농학회 2003 한국환경농학회지 Vol.22 No.3

Methoxyacrylate계 신규 살균제 KNF 1002의 오이, 고추 및 고춧잎에 대한 잔류 양상을 조사하기 위하여 포장시험을 수행하였다. HPLC-DAD에 의한 KNF 1002의 잔류분석시 최소검출량은 오이 및 고추에서 2.0 ng 그리고 고춧잎에서는 2.5 ng이었으며, 검출한계는 오이 및 고추에서 0.02 mg/kg 그리고 고춧잎에서 0.05 mg/kg 이었다. 오이에서의 잔류량은 1회 및 2회 처리하여 1일에서 7일후에 채취된 시료에서 2.0 mg/kg에서 검출한계 미만으로 나타났다. 고추에서의 잔류량은 시설재배지의 경우 최고 0.79 mg/kg에서 0.31 mg/kg 수준으로 나타났다. 노지재배의 경우에는 0.47 mg/kg에서 0.11 mg/kg으로 나타나 시설재배의 47% 수준의 잔류량을 보였다. 고춧잎에서는 시설재배지의 경우 최고 25.20 mg/kg에서 7.38 mg/kg으로 나타났으며 노지재배의 경우에는 1.99 mg/kg에서 0.11 mg/kg 수준으로 나타나 시설재배에 비해서 6.2%정도의 잔류량을 보였다. This study was conducted to evaluate the terminal residue of a new fungicide, KNF 1002, in cucumber and pepper under greenhouse and field conditions. When a microemulsion formulation (20%) of KNF 1002 was applied once or twice during 1-7 days before harvest, its terminal residue in cucumber ranged <0.02-0.20 ㎎/㎏ under greenhouse condition. In pepper, its figure recorded 0.31-0.79 ㎎/㎏ and 0.11-0.28 ㎎/㎏ under greenhouse and field conditions, respectively. Much higher level of terminal residues was observed in leaves than those in fruits in pepper, showing 7.38-25.20 ㎎㎏ and 0.11-1.99 ㎎㎏ under greenhouse and field conditions, respectively. Cultivation condition affected evidently the residue level in pepper harvests. Residual pattern of KNF 1002 seemed to be comparable to strobilurin fungicides currently used.

Investigation into effects of transgenic glufosinate-resistant Zoysia grasses with herbicide application on bacterial communities under field conditions

Lee, Yong-Eok,Lee, Sun Hwa,Ryu, Gil-Do,Kang, Hong-Gyu,Kwon, Yong-Ik,Sun, Hyeon-Jin,Park, Kee Woong,Lee, Bumkyu,Song, In-Ja,Lim, Pyung-Ok,Lee, Hyo-Yeon Springer-Verlag 2015 Journal of Plant Biology Vol.58 No.5

Thermoanaerobacterium saccharolyticum B6A-RI의 Endoxylanase에 대한 유전적 편성 및 분자구조 연구

Lee, Yong Eok 동국대학교 경주대학 1994 東國論集 Vol.13 No.-

Thermoanaerobacterium saccharolyticum B6A-RI로부터 cloning된 endoxylanase (xynA)와 β-xylosidase (xynB) 유전자들은 염색체 DNA내에서 서로 밀집되어 존재하였고, 같은 전사방향을 가지고 있었다. T. saccharolyticum B6A-RI의 xynA는 family F β-glycanases에 속하는 유전자들과 유사한 아미노산 서열을 가지고 있었다. xynA의 N-terminal에서 부터 1.75-kb를 줄인 결과, 활성에는 변화가 없으나 열에 대한 안정성이 떨어진 효소가 형성되었다. C-terminal로 부터 1.05-kb를 줄인 것은 활성이나 열 안정성에 영향을 주지 않았다. DNA 염기서열로부터 추정한 아미노산 서열을 바탕으로 endoxylanase의 분자구조를 분석한 결과 endoxylanase는 여러 domain의 복합구조이며 C-terminal 끝은 S-layer-like domain으로 구성되어 있음이 밝혀졌다. T. saccharolyticum B6A-RI의 endoxylanase는 세포 벽에 존재함이 알려져 있으므로 이 결과는 이 domain이 endoxylanase를 세포 표면에 부착시키는 역할을 하는 것으로 사료된다. Endoxylanase의 sequence 분석결과및 이전의 연구결과를 근거로 하여 endoxylanase가 S-layer에 붙는 model이 제안되었다. In the cloned chromosomal DNA from Thermoanaerobacterium saccharolyticum B6A-RI, the endoxylanase (xynA) and β-xylosidase (xynB) genes were clustered within a 20-kb region, and had the same transcriptional orientation. xynA from T. saccharolyticum B6A-RI had strong similarity to genes from family F β-glycanases. Deletion of 1.75-kb from the N-terminal end of xynA resulted in an enzyme that retained activity but lost thermostability. Deletion of 1.05-kb from the C-terminus did not alter thermostability or activity. The molecular structure of the endoxylanase was analyzed with the deduced amino acid sequence. Sequence analysis revealed a composite structure of the endoxylanase consisting of several domains. The C-terminal end consisted of S-layer-like domain. Since the endoxylanase of T. saccharolyticum B6A-RI is known to be cell bound, this result suggests that this domain might serve as an anchor which mediate the attachment of the endoxylanase to the cell surface of this organism. Based on sequence analysis of the endoxylanase and results from previous studies, a model for the attachment of the endoxylasase to S-layer is proposed.

Expression of Human Lactoferrin Gene in Transgenic Rice (Oryza sativa L.)

Lee, Yong-Eok,Oh, Seong-Eun,Nishiguchi, Satoshi,Riu, Key-Zung,Song, In-Ja,Park, Shin-Young,Lee, Jin-Hyoung,Kim, Il-Gi,Suh, Suk-Chul,Rhim, Seong-Lyul,Lim, Pyung-Ok,Lee, Hyo-Yeon The Korean Society of Plant Biotechnology 2007 식물생명공학회지 Vol.34 No.2

Lactoferrin is an 80-kDa iron-binding glycoprotein known to exert many biological activities, such as facilitating iron absorption and having antimicrobial and anti-inflammatory effects. Rice can be a useful target for edible food plants to introduce human lactoferrin, because it has lower allergenicity and is likely to be safer than microorganisms or transgenic animals. A cDNA fragment encoding human lactoferrin (HLF) driven by the maize polyubiquitin promoter, along with herbicide resistance gene (bar) driven by CaMV 35S promoter, was introduced into rice (Oryza sativa L. cv. Dong Jin) using the Agrobacterium -mediated transformation system. Putative transformants were initially selected on the medium containing bialaphos. The stable integration of the bar and HLF genes into transgenic rice plants was further confirmed through polymerase chain reaction (PCR) and Southern blot analyses. The expression of the full length HLF protein from various tissues such as grains and young leaves of transgenic rice was verified by Western blot analysis. Analysis of progeny also demonstrated that introduced genes were stably inherited to the next generation at the Mendelian fashion.

Hemicellulose분해 박테리아인 Thermoanaerobacteriumsaccharolyticum B6A-RI의 세포 표면구조에 대한 초미세 구조학적 연구

Lee, Yong-Eok 동국대학교 경주대학 1994 東國論集 Vol.13 No.-

이 논문은 hemicellulose분해 박테리아인 Thermoanaerobacterium saccharolyticum B6A-RI의 세포 표면성분에 대한 초미세 구조학적 연구이다. Xylan 또는 xylose에서 자란 T. saccharolyticum B6A-RI의 세표 표면구조를 cationized ferritin과 gold labeling방법을 이용하여 전자 현미경으로 관찰하였다. 이들 구조들은 glucose에서 자란 세포들에서는 나타나지 않았다. 이 구조들 (xylanosome)의 형성은 endoxylanase의 생성 및 박테리아들이 xylan에 흡착하는 현상과 연루되어 있다. 이 xylanosome은 xylan에만 작용하며 cellulose에는 흡착하지 않는다. 이러한 발견은 박테리아의 세포벽에 대한 지식을 확장시키며, 또한 세포표면구조가 세포가 xylan에 흡착하여 이를 이용하는데 관여한다는 증거를 제시한다. 이러한 hydrolysome구조들은 미생물들이 불용성기질을 효과적으로 분해하여 세로 내로 운반하기 위한 glycoprotein complex를 형성함을 나타낸다. This report provides an ultrastructural study of the cell surface components of hemicellulolytic bacterium, Thermoanaerobacterium saccharolyticum B6A-RI. Negatively charged surface structures were visualized on cells of T. saccharolyticum B6A-RI grown on xylan or xylose, using cationized ferritin and gold labeling with scanning and transmission electron microscopy. These structures were absent on glucose grown cells that were non-xylanolytic. The presence of these putative xylanosome structures coincided with production of cell bound endoxylanases and the ability of the cells to bind tightly to xylan. The xylanosoms appeared to be specific for cellular adhesion to xylan, as cells would not bind to cellulose. These finding extend the knowledge about the surface structures known the exist on the bacterial cell wall, and provide the first example of a cell surface structure specific for cellular adhesion to and degradation of xylan. These hydrolysome structures further demonstrate the ability of microorganisms to effectively form a glycoprotein complex to efficiently degrade insoluble substrates by facilitating both the concerted action of hydrolytic enzymes and the transfer of degradation products into the cell.

Isolation of a Thermophilic Bacterium Producing a Thermostable a-Glucosidase and Characterization of the Enzyme

Lee, Yong-Eok 동국대학교 생명과학연구소 1995 생명과학연구논문집 Vol.2 No.-

A thermophilic bacterium producing a thermostable α-glucosidase was isolated from manure and as been identified as Bacillus sp. The optimum temperature for growth was 65°C. No growthwas obtained at 40 or 75°C. The optimum pH for growth was 5.5 to 8.5. The α-glucosidase production was constitutive in the absence of carbohydrates. High level of enzyme activity was detected during growth in starch. Addition of glucose resulted in the repression of the α-glucosidase production. The optimum pH and temperature for enzyme activity were pH 4.8 to 5.0 and 60 to 65°C, respectively.