Cyclodextrin Glucanotransferase와 Cyclodextrinase를 생산하는 Bacillus 속 세균의 분리와 그 효소들의 특성

권현주,남수완,김광현,곽영규,김병우 동의대학교 기초과학연구소 1997 基礎科學硏究論文集 Vol.7 No.1

A bacterium producing Cyclodextrin Glucanotransferase (CGTase) and Cyclodextrinase (CDase) was isolated from soil, and named as Bacillus stearothermophilus KJ16 . The growth of the isolated strain occurred in two steps, and syntheses of CGTase and CDase were dependent on the growth cycle of the cell. CGTase was constitutively synthesized during the 1st growing phase. while CDase was synthesized inducibly during the 2nd growing phase. When the medium pH was controlled at 7.0, the maximun enzyme activities of CGTase and CDase were increased by 12-fold (1300 mU/ml) and 2-fold (225 mU/ml), respectively, compared with the pH-uncontrolled batch culture. The CGTase of the isolate converted soluble starch to CDs with the ratio of α-CD:β-CD:γ-CD=42:46:12 at 55℃. The optimal pH and temperature of CGTase were 6.0 and 60℃,respectively and the optimal pH and temperature of CDase were 6.0 and 55℃. The molecular weights of the purified CGTase and CDase were estimated to be 65,000 and 68,000 dalton, respectively, Among serveral substrates, γ-CD was most rapidly hydrolyzed by the purified CDase.

간암세포에서 저산소증에 의해 유도되는 VEGF의 발현기작에 대한 연구

권유욱,배수경,김정애,김규원,박병채 부산대학교 유전공학연구소 1997 분자생물학 연구보 Vol.13 No.-

Purpose: Hepatocellular carcinoma(HCC),a typical hypervasculized tumor is very sensitive to hypoxia and vascular endothelial growth factor(VEGF) has previously been identified to be up-regulated in response to hypoxia in several cell types. However, the molecular mechanisma by which hypoxia is sensed by the cells remain enigmatic. To investigate whether calcium and AP-1 are involved in hypoxia-sensing mechanism, we performed following eaperiments. Materials and Methods: Hep3B cells were grown in hypoxia condition. To assess cell viability, MTT assay was performed. To investigate the effect of calcium and AP-1,northern blot analysis was performed after treatment with BAPTA/AM. Results: The expression of VEGF was significantly up-regulated by hypoxia in Hep3B, hepatocellular carcinoma cell line. The increased expression of VEGF induced by hypoxia was blocked by the addition of BAPTA/AM, a cytosolic calcium chelator to the media. In addition, we found that the expression of c-jun protooncogene was also-regulated by hypoxia. Hypoxia increase of c-jun expression was also normalized by the treatment with BAPTA/AM. Conclusion: These results suggest that the increased expression of VEGF by hypoxia is mediated through the calcium and c-jun signalling pathway in the Hep3B human hepatoma cell lines.

하수슬러지의 건설재로의 이용 타당성에 관한 연구

권재혁,박병수 三陟大學校 1999 論文集 Vol.32 No.2

Feasibility of using the mixture of a wastewater sludge and additive as liner and the cover materials in disposal landfill. Granite weathered soil flyash, produced as a by-produced as a by-product in the power plant, were used as the primary additives to improve geotechnical engineering properties of sludge. For secondary additives, bentonite and cement were mixed with sludge to decrease the permeability and to increase the shear strength, irrespectively. Various kinds of laboratory test, required to evaluate the design criteria for the liner and the cover materials, were carried out by changing the mixing ratio of sludge with the additives. Basic soil property tests of specific gravity, grain size distribution, the liquid and the plastic limits were performed to evaluate their properties and to analyze their effects on permeability, compaction of mixtures. Various test results with mixtures were assessed to evaluate whether their properties met the requirements as the liner and the cover materials in waste disposal landfill.

간암세포에서 저산소증에 의해 유도되는 VEGF의 발현기작에 대한 연구

권유욱,배수경,김정애,김규원,박병채 영남대학교 약품개발연구소 1997 영남대학교 약품개발연구소 연구업적집 Vol.7 No.-

Puropse: Hepatocellular carcinoma (HCC), a typical hypervasculized tumor is very sensitive to hypoxia and vascular endothelial growth factor (VEGF) has previously been identified to be up-regulated in response to hypoxia in several cell types. However, the molecular mechanisms by which hypoxia is sensed by the cells remain enigmatic. To investigate whether calcium and AP-1 are involved in hypoxia-sensing mechanism, we performed following experiments. Materials and Methods: Hep3B cells were grown in hypoxic condition. To assess cell viability, MTT assay was performed. To investigate the effect of calcium and AP-1, northern blot analysis was performed after treatment with BAPTA/AM. Results: The expression of VEGF was significantly up-regulated by hypoxia in Hep3B, hepatocellular carcinoma cell line. The increased expression of VEGF induced by hypoxia was blocked by the addition of BAPTA/AM, a cytosolic calcium chelator to the media. In addition, we found that the expression of c-jun protooncogene was also up-regulated by hypoxia. Hypoxic increase of c-jun expression was also normalized by the treatment with BAPTA/AM. Conclusion: These results suggest that the increased expression of VEGF by hypoxia is mediated through the calcium and c-jun signalling pathway in the Hep3B human hepatoma cell lines.

Cyclodextrin Glucanotransferase와 Cyclodextrinase를 생산하는 Bacillus 속 세균의 분리와 그 효소들의 특성

권현주,남수완,김광현,곽영규,김병우 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.3

토양으로부터 CGTase와 CDase를 함께 분비·생산하는 내열성 세균을 분리하였으며, 동정결과 Bacillus stearothermophilus로 판명되어 KJ16으로 명명하였다. Bacillus stearothermophilus KJ16 균주는 회분배양할 경우 2단계 성장을 하며 일차증식기에 CGTase를 생산하고 이차증식기에 CDase를 생산하는 양상을 보였다. 배치 pH를 7.0으로 조절하면서 배양할 경우, CGTase는 균의 증식에 따라 증가하고 CDase는 CGTase보다 뒤에 생산되었다. 그 생산양은 pH 비조절 배양에 비해 CGTase는 최대 12배(1300 mU/ml)까지, CDase는 2배(225 mU/ml)정도 증가되었다. 분리균주가 생산하는 CGTase는 분자량이 65,000 dalton으로 pH 6.0, 60℃에서 최적활성을 보이며 HPLC 분석을 통해 α-, β-, γ-CD를 42: 46: 12의 비율로 생산하는 α/β 혼합형 CGTase였다. 또한 CDase는 분자량이 68,000 dalton으로 pH 6.0, 55℃에서 최적활성을 보이며 최적기질은 γ-CD였다. A bacterium named as Bacillus stearothermophilus KJ16. The growth of the isolated strain occurred in two steps, and syntheses of CGTase and CDase were dependent on the growth cycle of the cell. CGTase was constitutively synthesized during the 1st growing phase, while CDase was synthesized inducibly during the 2nd growing phase. When the medium pH was controlled at 7.0, the maximun enzyme activities of CGTase and CDase were increased by 12-fold(1300 mU/ml) and 2-fold (225 mU/ml), respectively, compared with the pH-uncontrolled batch culture. The CGTase of the isolate converted soluble starch to CDs with the ratio of α-CD: β-CD: γ-CD=42:46:12 at 55℃. The optimal pH and temperature of CGTase were 6.0 and 60℃, respectively and the optimal pH and temperature of CDase were 6.0 and 55℃. The molecular weights of the purified CGTase and CDase were estimated to be 65,000 and 68,000 dalton, respectively. Among serveral substrates, γ-CD was most rapidly hydrolyzed by the purified CDase.

Bacillus stearothermophilus KJ16이 생산하는 Cyclodextrin Glucanotransferase의 정제와 효소특성

김병우,김광현,남수완,권현주,송승구,윤종원 동의대학교 기초과학연구소 1999 基礎科學硏究論文集 Vol.9 No.1

Cyclodextrin glucanotransferase from B. stearothermophilus KJ16 that can produce both cyclodextrin glucanotransferase and cyclodextrinase was purified by ammonium sulfate precipitation, DEAE-cellulose chromatography, Sephadex G-100 chromatography, and FPLC. The molecular weight of the purified enzyme was about 65,000 dalton by SDS-PAGE. The optimal pH and temperature were 6.0 and 60℃, respectively. The enzyme was stable at 50℃ for 1 hr and in the pH range of 5.5 and 8.5. Mercaptoethanol and dithiothreitol inhibited the enzyme activity strongly. The enzyme produced 60% cyclodextrin(CD) from 5% soluble starch with the ^α-, ^β-, ^γ-CD ratio of 42 : 46 : 12. Amylopectin was the most suitable with 67% conversion to CD.

단일전력단으로 구성된 AC/DC 풀 브리지 컨버터에 관한 연구

안병무,권순도,김용,백수현 동국대학교 산업기술연구원 2000 산업기술논문집 Vol.11 No.1

본 논문에서는 단일단 AC/DC 폴브리지 토포로지에 적합한 고주파 소프트 스위칭 컨버터응용에 근거를 두고 제안한다. 제안된 컨버터는 입력전류를 불연속 모드로 동작시켜 역률을 개선하며 입력전류의 zero-crossing-point에서의 왜곡을 개선함으로써 고조파를 감소시켜 고역률, 영전압 스위칭, 저잡음과 고효율을 구현한다. 펄스폭 변조 제어방식은 각각의 스위칭과 정류손실을 저감하기 위해 사용되었다. 제안한 컨버터는 IGBT를 전력스위치로서 주로 사용되지는 고전압, 고전력 컨버터응용에 적용할 수 있으며, 실험을 통하여 1.5kW, 30kHz로 동작하는 컨버터를 구현하였다. A single stage AC/DC converter based on a full bridge topology suitable for high frequency soft switching converter applications is proposed. The power factor correction can be achieved by the discontinuous conduction mode(DCM) operation of the input current. The proposed converter gives the good power factor, low line current harmonics. The proposed converter has high power factor, zero voltage switching low noise and high efficency. A pulse width modulation control is employed to reduce the switching and rectification losses respectively. The proposed converter is attractive for high-voltage, high-power applications where IGBT's are predominantly used as the power switches. The principle of operation, features, and design are verified on a 1.5kW, 30kHz, IGBT based experimental circuit.

새로운 Baculoviurs 전이벡터를 이용한 Escherichia coli β-galactosidase 유전자의 발현

우수동,김우진,김혜성,진병래,강석권 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1

국내에서 분리된 BmNPV를 이용하여 제작된 새로운 전이벡터 pBmKSK1에 외래 유전자로서 E. coli lacZ 유전자를 클로닝하고 재조합 바이러스를 제작하였다. 재조합 바이러스에 대하여 Southern blotting 분석을 실시하여 lacZ 유전자의 존재를 확인하였으며, 재조합 바이러스가 접종된 세포의 SDS-PAGE 및 Western blotting 분석을 통하여 β-galactosidase의 발현을 확인하였다. 재조합 바이러스의 발현효율을 누에 세포주 및 유충에서 비교 조사한 결과, 유충에서 더욱 높은 발현량을 보임을 확인하였다. To investigate the expression efficiency of new transfer vector of Bombyx mori nuclear polyhedrosis virus (BmNPV), Escherichia coli lacZ gene was inserted into new transfer vector pBmKSK1, under the control of polyhedrin promoter and expressed in BmN-4 cells coninfected with transfer vector pBmKSK1-LacZ and vild type BmNPV genome, and analysed by Southern blotting. The expression of β-galactosidase was characterized by SDS-PAGE, Western blotting and β-galactosidase activity assay. The results showed that the level of expression in silkworn larvae was higher than that of BmN-4 cells.

蒐集 在來種 고추의 特性(1988年度)

金炳洙,李愚升,權寧石,孫銀鈴 慶北大學校 1989 論文集 Vol.48 No.-

Twenty three local cultivars of pepper(capsicum annuum L.) were planted in the Kyungpook National University Experiment farm and their growth and fruit characteristics were investigated. Seedings grown separately were tested for resistance to bacterial spot(Santhomonas campestris pv. vesicatoria) and Phytophthora blight(Phytophthora capsici) by artificial inoculation. KC209 was the earliest in flowering followed by KC218, KC203, KC223, KC202, KC201 and KC200. KC213, KC217, and KC204 were leading in fresh red fruit yield in a descending order among the rest excluding KC205 and KC207, open-pollinated fruits of which were removed and flower buds were wrapped before blooming to guarantee self-pollination. Fruits of Kc207 was the largest and Kc221 and KC217 were also bearing relatively large iruits. None of the cultivars tested was resistant to either bacterial spot or Phytophthora blight. Collection of local cultivars of pepper was continued in 1988 and 7 local cultivars were collected in Kyungpook, Cheonbuk, and Cheonnam provinces and their seed fruit characters were recorded.

유전자 증폭법(PCR)을 이용한 methicillin-registant Staphylococcus 균속의 신속동정에 관한 연구

김영권 ; 장철수 ; 김병원 ; 박영진 김천대학교 2000 김천대학교 논문집 Vol.21 No.-

In order to indentify rapidly the methicillin-resistant genus Staphylococcus (MRS) which is known as a causative bacteria of nosocomial infection, we performed the polymerase chain reaction (PCR) method. In this study, we used 44 strains which were consist of 39 starins of Staphylococcus aureus, 2 strains of Staphylococcus epidermidis. 1 strain of Staphylococcus warneri, 1 strain of Staphylococcus cohnii isolated from clinical specimens and 1 type strain of Staphylococcus aureus KCTC 1928 for PCR experiments. DNA was extracted directly from the bacterial colony. 22 mer-oligonucleotides were used as primer which complements to the mecA region of structural gene coding low affinity binding pro-tein (PBP2a). A single band of 533 bp length from MRSA, MRSE and MRSW were confirmed. Also, an antibiotic susceptibility test by agar dilution method of NCCLS was performed for the strains isolat-ed from clinical specimens. The results obtained by this method were compared with the PCR results. In the result, most of the strains which have strong resistance to methicillin were amplified for the 533 bp fragment of the mecA gene in PCR experiment. However, 4 strains (MRA1-10, MRW2-3. MRE1-15. MRE1-16) that were classified as a sensitive group in the antibiotic susceptibility test also was showed mecA gene fragment in PCR. This suggested that the specificity of mecA gene is not always consistant with that of MRSA. In order to confirm the fragment in a region of mecA gene, a southern blot hybridization process was performed using the PCR products as probe. As a result, a single hybrid band was detected at about 4 Kb region of Hind III and Hae III-digested DNA fragment in MRS. Conclusively for the diagnosis of MRS, the confirmation of mecA gene by PCR seems to be more accurate and rapid than the antibiot-ic sensitivity test.