제조합 대장균에서 과발현된 Citrobacter freundii KCTC2006 유래의 β-Tyrosinase를 이용한 3,4-Dihydroxyphenyl-L-alanine의 생산

이승구,노현수,홍승표,이규종,왕지원,태동년,엄기남,방상구,김영준,성문희 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1

재조합 대장균에서 대량발현 시킨 Citrobacter freundii KCTC 2006 유래이 효소 β-tyrosinase를 이용하여 pyrocatechol, sodium pyruvate, ammonium acetate로부터 3,4-dihydroxy phenyl-L-alanine을 생산하기 위한 연구를 수행하였다. 이 효소반응에 적합한 온도 및 pH 조건은 각각 18℃와 8.5로 결정되었고, 반응액 중의 ammonium acetate와 sodium pyruvate의 농도는 각각 300 mM, 50 mM 이상으로 조절하는 것이 적합하였다. Pyrocatechol의 경우는 20 mM에서 가장 높은 반응성을 나타냈으나, 기질을 반복적으로 첨가하며 장시간 동안 효소반응을 수행하는 경우에는 pyrocatechol의 고갈을 피하기 위하여, 20 mM에서 50 mM 사이로 조절하였다. 한편, 반응액 중에 ethanol을 10% 첨가한 경우에는 반응속도가약 20% 증가하였다. 이상과 같은 효소반응특성에 기초하여 조제한 기질용액에 β-tyrosinase를 1 unit/㎖ 농도로 가하고, pyrocatechol과 pyruvate가 고갈되지 않도록 간헐적으로 첨가하면서 효소반응을 수행한 결과, 24시간 만에 85.2%의 수율로 31.6g/ℓ의 3,4-dihydroxyphenyl-L-alanine를 생산할 수 있었다. By using the β-tyrosinase of Citrobacter freundii KCTC2006, which was cloned and overexpressed in Escherichia coli, 3,4-dihydroxy phenyl-L-alanine (L-DOPA) was synthesized efficiently from pyrocatechol, sodium pyruvate, and ammonium acetate. Optimal temperature and pH for the reaction were determined to be about 18℃ and 8.5, respectively. The effects of substrate concentrations were also examined at different concentrations of ammonium acetate, sodium pyruvate, and pyrocatechol. Ammonium acetate and sodium pyruvate increased the reaction rate until the concentrations reached to 300 mM and 50 mM, respectively. Although pyrocatechol showed the optimal concentration at 20 mM, it was controlled between 20 mM and 50 mM to avoid the depletion of substrate during the enzymatic synthesis. Based on above results, a reaction medium for the production of L-DOPA was prepared and incubated with 1 unit/㎖ of β-tyrosinase. Pyrocatechol and sodium pyruvate was added to the reaction solution intermittently to avoid the substrate depletion during the enzymatic reaction. After 24 hour of reaction, 31.6 g/ℓ L-DOPA was accumulated in the reaction solution as soluble and precipitated ones and the conversion yield was about 85.2%.

제주도 고등학교 학생들의 비만 유병률과 심혈과 위험인자

황승욱,김현주,홍성철,이상이 제주대학교 생명과학연구소 2002 제주생명과학연구 Vol.5 No.6

연구배경 : 최근 청소년 연령층의 비만이 급격히 증가하고 있으며, 비만 청소년들에 있어서 고혈압, 당뇨, 고지혈증이 조기에 발현될 뿐 아니라 성인비만과 성인병 발생의 주 원인이 되고 있다. 본 연구는 제주도내 고등학생들의 비만 유병률과 청소년 비만과 심혈간 위험인자인 혈압, 총콜레스테롤, 공복시혈당의 관련성을 알아보고 소아비만과 청소년 비만의 관련성을 조사하였다. 방법 : 연구대상은 제주도내 22개 고등학생 1학년 6,064명을 대상으로 하였고. 체격조사는1999년도 학교 신체검사 자료와 병원 건강검진 자료를 이용하였다. 비만에 대한 평가는 대한소아과학회에서 발행한 1998년도 한국소아표준치와 체질량 지수 두가지를 이용하였다 심혈관 위험인자로는 총콜레스테롤 수축기 혈압, 이완기혈압 공복시 혈당으로 하였으며, 배상자중 646명에 대해 과거 건강기록부를 열람하여 초등학생 때의 비만실태를 조사하였다. 결과 : 제주도 고등학생의 비만 유병률은 표준체중에 의한 비만 산출법으로는 남, 여학생 각각 12.6%, 13.3%이었고 체질량 지수에 의한 비만 위험군 및 비만 학생이 남, 녀 각각 12.7%, 13.4%로 나타났다. 지역에 따른 비만 유병률은 남학생의 경우 시 지역 12.9%, 군 지역 10.3%로 시 시역에서 비만 유병률이 유의하게 높았으며, 여학생의 경우 시·군지역 각각 12.9%, 14.2%이었으나 통계적 유의성은 없었다. 고등하교 계열에 따른 비만 유병률은 남학생의 경우 인문계 13.9%, 실업계 10.1%로 인문계가 유의하게 높았으나, 여학생의 경우는 통계적 유의성은 없었다. 비만 정도에 따른 심혈관 위험인자와의 관련성에서 남학생의 경우 수축기와 이완기혈압, 공복시혈당, 총콜레스테롤 수치가 비만 정도에 따라 유의한 차이를 보였으며(P<0.05), 여학생의 경우 공복시혈당을 제외한 나머지 변수에서 유의한 차이를 보였다. 체질량지수와 심혈간 위험인자간의 상관관계에서 남학생은 혈압, 공복시혈당 총콜레스테롤이 유의한 양의 상관관계를 보였으며, 여학생의 경우 공복시혈당 외에 다른 수치에서 유의한 양의 상관관계를 보였다. 연구대상자 중 646명에 대해 초등학교 1학넌 때의 비만실태를 추적한 결과 초등학생 비만이 고등학생 비만에 미치는 비교위험도가 남녀 각각 3.77, 2.16으로 유의하게 나타났다(P<0.05). 결론 : 제주도지역 고등학생들의 비만 유병률이 상당히 높게 나타났고, 청소년 비만과 심혈관 위험인자간의 관련성을 알 수 있었다. 또한 소아비만이 청소년 비만으로 상당수 이행되었으므로 소아 및 청소년기에 적절한 비만관리 프로그램이 요구된다. Background: Recently, adolescent obesity was increased and tended to become obese adults, facing increased risk for diabetes, cardiovascular disease. Even vefore reaching adulthood, adolescent, obesity already was experiencing hypertension, dyslipidemia, and diabetes mellitus. The purpose of this study was to find prevalence of obesity of high school students in Jeju island and association between obesity and cardiovascular risk factors and association between childhood obesity and adolescent obesity. Method: The study subjects were 6,064 students of twenty-two high school in Jeju island and health survey data was gathered from high school health examination data and hospital health examination data from Aug. 1999 to Dec. 1999. Adolescent obesity was evaluated by ideal body weight method in 1998 and body mass index. Cardiovascular risk factors were checked total cholesterol, systolic and diastolic blood pressure, fasting blood sugar. In 646 students, obesity at primary school is checked by primary school health record. Results: Prevalence of obesity in Jeju island high school students was 12.6% in male students, 13.3% in female students by ideal body weight method and Prevalence of obesity risk and obesity was 12.7% in male students, 13.4% in female student by body mass index. Prevalence of obesity of male students in city area and district area of Jeju island were each 12.9%, 10.3% and that of male students in city area was significantly higher. Prevalence of obesity of female students in city area and district area of Jeju island were each 12.9%, 10.3% and that of male students in city area was significantly higher. Prevalence of obesity of female students in city area and district area of Jeju island were each 12.9%, 14.2% and not significantly different. Prevalence of obesity of male students in general and commercial school group were each 13.9%, 10.1% and that of general school group was significantly higher. In female students, Prevalence of obesity general and commercial school group was not significantly different. Total cholesterol, systolic and diastolic blood pressure were significantly associated with degree of obesity in male and female students(P<0.05) and fasting blood sugar was only significantly associated with degree of obesity in male students (P<0.05). In 646students, Relative risk of obesity at primary school and that at high school in male and female students were 3.77, 2.66 and was significantly associated. Conclusion: Prevalence of obesity of high school students in Jeju island is very high and adolescent obesity was significantly associated with cardiovascular risk factors. Mostly childhood obesity tended to become adolescent obesity and program of control of obesity at childhood and adolescence is required.

노인 입원 환자의 질병 양상과 의료이용 실태

김현주,홍성철,이상이,김명주,황승욱 대한노인병학회 2003 대한노인병학회지 Vol.7 No.4

Trichloroethylen의 직업적 노출에 의한 독성간염 및 박탈성 피부염 1례

채홍재,이성관,이강진,김종율,이승철,신동현,문재동 大韓産業醫學會 2003 대한직업환경의학회지 Vol.15 No.1

28세 남자가 trichloroethylene을 이용한 세척 및 탈지작업에 종사한지 한 달만에 발열, 오한 및 홍반을 동반한 피부발진 등의 증상이 발현되었다. 황달을 동반한 간기능 부전의 소견을 보였으며 피부는 박탈되었다. 부신피질호르몬, 항히스타민제 및 간기능 보조제를 이용한 치료로 간기능 및 피부 병변은 호전되었다. 검사소견상 Ig E가 상승되었고 trichloroethylene에 의한 과민반응으로 추정되었다. Trichloroethylene을 사용하는 소규모 사업장에서의 노출억제 방안을 포함한 보건관리가 요구되며 특수건강진단 주기에 대한 변경이 필요하리라 사료된다. A 28-year-old male began working as a degreaser. The solvent used in the degreasing operation was trichloroethylene. Over the next month the man experienced fever, chills, and an erythematous skin rash and itching. At that time he had a marked elevation in his liver enzyme, with cholestasis. Over the next few days the rash persisted then peeled. There was an elevation of Ig E, and a positive patch test reaction to trichloroethylene. His dermatitis and hepatitis were considered to be mediated by a hypersensitivity mechanism.

3,4-Dihydroxyphenyl-L-alanine의 효소적 생산에 대한 반응첨가물의 영향

이승구,노현수,홍승표,성문희 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.2

재조합 대장균에서 과발현된 Citrobacter freundii KCTC 2006 유래의 tyrosine phenol-lyase(E.C.4.1.99.2) 이용 3,4-dihydroxyphenyl-L-alanine (L-DOPA)의 생산에 대한 반응첨가물의 영향을 조사하였다. 반응액 중 효소 및 조효소 농도의 영향을 조사한 결과, 효소농도는 약 2 units/ml이 적합하였으며, 조효소인 pyridoxal -5-phosphate 는 0.1 mM 이상이 필요하였다. Pyrocatechol과 결합하여 안정한 복합체를 형성하는 sodium borate는 pyrocatechol에 의한 효소의 불활성화를 감소시키는 효과가 있었으나, 효소의 반응성을 현저히 저하시켜 L-DOPA 생산의 관점에서는 불리한 단점도 있었다. 한편, 알콜류 등의 유기용매가 L-DOPA 합성반응에 미치는 영향을 조사한 결과 methanol을 5% 농도로 반응액에 첨가하였을 때, 효소의 반응과 안정성이 크게 증가하여 L-DOPA 합성반응이 지속적으로 수행됨으로써 고농도의 L-DOPA를 효율적으로 생산할 수 있게 되었다. 생산된 L-DOPA의 약 77%가 불용성 상태로 침전되어 쉽게 회수할 수 있었으며, 침전된 L-DOPA를 1N HCI에 용해한 후 재결정화 함으로써 최종적으로 99.96%의 고순도 L-DOPA를 생산할 수 있었다. The enzymatic synthesis of 3,4-dihydroxyphenyl-L-alanine (L-DOPA) was examined for the effects of the reaction additives such as sodium borate, alcohol, and organic solvents. The enzyme used was tyrosine phenol-lyase of Citrobacter freundii KCTC 2006 produced in Escherichia coli. The amounts of tyrosine phenol-lyase and pyridoxal -5-phosphate were optimized to 2.0 units/ml and 0.1 mM, respectively, for the synthetic reaction. Sodium borate, a substance that forms a complex with pyrocatechol, reduced the enzyme deactivation by pyrocatechol although it seriously inhibited the enzyme activity. Among the organic solvents tested, dimethylsulfoxide, dimethylformamide, and alcohol increased the productivity of the L-DOPA synthesis. In a reaction system with 5% methanol, L-DOPA concentration increased up to 210 mM after 24 hours, and 77.1% of which was separated as precipitates. The L-DOPA was purified to 99.96%

가토에서 음각틀을 이용한 이개 재건의 실험적 연구

민대홍,김승홍,백무현,김원석 大韓成形外科學會 1991 Archives of Plastic Surgery Vol.18 No.4

The formation or reconstruction of an auricle is one of the most difficult operations in plastic and reconstructive facial surgery. The difficulties of its reconstruction include delicate structures of the ear, extremely thin skin, difficulty in color match, and springing effect of the thin supporting skeleton consists of cartilage with its peculiar recesses and bumps. Even though, several techniques have been using for auricular reconstruction, it would be difficult to obtain an ideal auricle. Furthermore surgeon's talent and sufficient practice might be needed for a satisfactory result. We attempted auricular reconstruction using negative auricular plastic mold in 10 white rabbits in order to pursue a simple and satisfactory method. Diced cartilage chips were filled into a subcutaneous pocket placed in the negative auricular mold. Mild negative pressure was maintained in the space between the cartilage-subcutaneous tissue-skin complex and mold for two months. The results were as follows: 1.An ideal shaped suitable supporting skeleton was newly formed and the fabricated new auricle had a near normal appearance. 2.Under the microscopic examination, the diced cartilage chips have conglomerated together to form a solid skin - connective tissue - cartilage framework and evenmore matured chondrocytes were noted.

강원도산 참당귀와 일본산 일당귀의 생리 활성 성분 탐색

함문선,김성수,홍종수,이진하,정을권,박영식,이현용 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.5

참당귀와 일당귀를 에탄올로 추출하였고 이들 추출물들을 디에틸 에테르층과 물층으로 분획하였다. 두 개의 분획물들은 Ames mutagenicity test에서 MNNG(0.5 ㎍/plate)에 대한 강한 항돌연변이 효과를 나타내었다. 디에틸 에테르 분획물들은 MNNG의 돌연변이원성을 78%에서 80%까지 억제시켜 가장 강한 돌연변이 유발 억제효과를 보여주었다. 각각의 에탄올 추출물(1㎍/㎕)들은 사람의 다양한 암세포들의 성장을 억제시켰다. 특히, 디에틸 에테르 분획물들은 간암 세포의 생육을 90%에서 95%까지 억제시켜 가장 강한 항암효과를 보여주었다. 그러나 수층 분획물들은 암세포의 생육에 대하여 거의 억제활성을 나타내지 않았다. 에탄올 추출물들은 인체의 정상 간세포에 대하여 거의 세포독성을 나타내지 않았다. 참당귀의 디에틸 에테르 분획물들은 일당귀와 비교하여 20% 미만의 세포독성을 보여주었다. 참당귀와 일당귀 디에틸 에테르 분획물들의 0.5㎍/㎕ 첨가는 T세포와 B세포의 생육의 3~4배 정도 증가에 의한 면역활성을 보여주었다. 또한 참당귀의 디에틸 에테르 분획물(0.7㎍/㎕)이 ACE 활성을 98%까지 억제시켜 혈압을 조절할 수 있음도 증명되었다. TLC로부터, 두 개의 디에틸 에테르 분획물들은 참당귀와 일당귀의 활성 성분중의 하나로 알려진 umbelliferon을 함유함을 알 수 있었다. The ethanol extracts from Anglica gigas Nakai and Angelica acutiloba Kitagawa were fractionated to diethyl ether and aqueous partitions. Both partitions had strong antimutagenic effect on the MNNG (N-methyl-N-nitro-N-nitrosoguanidine) by Ames mutagenicity of MNNG with inhibition of 78~80%. The ethanol extracts from both species showed the inhibitory effect on the growth of several human cancer cell lines. Especially, the diethyl ether fraction from ethanol extracts was most effective on human hepatocellular carcinoma cells, inhibiting 90~95% of cell growth. However, the aqueous fractions had least inhibition activity on many cancer cells. There was little cytotoxicity on human normal liver cell by ethanol extracts. Diethyl ether fraction from Angelica gigas Nakai ethanol extract had cytotoxicity les than 20% on human normal liver cells, compared with that from Angelica acutiloba Kitagawa ethanol exract. The adding of 0.5 (g/ℓ) of diethyl ether fractions of Angelica gigas Nakai or Angelica acutiloba Kitagawa increased immune activity by enhacing human B and T cells up to three to four times. It was proven that diethyl ether fraction (0.7 g/ℓ) from Angelica gigas Nakai could control blood pressure by suppressing angiotensin converting enzyme activity up to 98%. From TLC, it was appeared that both of diethyl ether partitions had umbelliferon, known to one of active substances from Angelica gigas Nakai and Angelica acutiloba Kitagawa.

원격탐사 기법을 이용한 도시토지환경 분석에 관한 연구

조영환,송태갑,홍현승 木浦大學校 沿岸環境硏究所 1998 沿岸環境硏究 Vol.15 No.1

This study compares the usefulness of remote sensing with that of aerial photography as a main data source for urban land analysis and development planning. The data was collected over inland districts under the administrative control of Mokpo city, but the results of the study are easily generalized. The main environmental factors considered were annual changes in vegetative ground covering, categorized into sectors according to the type of surface soil supporting it. Remote sensing techniques facilitated the collection of large amounts of data over a wide area in a relatively short time. Remote sensing data was also easy to collect repeatedly on an ongoing basis, making it useful as a fundamental data source for town planning and urban land development. On the other hand, aerial photographs offer a superior spatial perspective on urban lands, making them very useful as data for detailed planning in specific areas slated for intensive development.

봉독약침이 류마티스 관절염 환자의 관절 통증, 종창 및 급성 염증 반응에 미치는 영향

이상훈,이현종,백용현,김수영,박재경,홍승재,양형인,김건식,이재동,최도영,이두익,이윤호 WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 2003 東西醫學硏究所 論文集 Vol.2003 No.-

Objective In order to study the effects of bee venom(BV) on the pain, edema, and acute inflammatory reactant of rheumatoid arthritis(RA) patients. Methods Patients with RA who met the ACR(American College of Rheumatology) 1987 revised criteria for the diagnosis of RA were treated with the BV therapy twice a week for 3 months. Tender Joint counts, swollen joint counts, Visual analog scale(VAS), morning stiffness, ESR. C-reactive protein(CRP) were analyzed before and after BV therapy. Results The results as follows. 1. Tender joint counts in patients after BV therapy were significantly lower than those before BV therapy(9.0±7.9 vs 15± 11.4, p=0.002). 2. Swollen joint counts of the patients after BV therapy were significantly lower than those before BV therapy(50±61 vs 15±23, p=0.001). 3. VAS in patients after BV therapy was significantly lower than those before BV therapy(608± 17.6 vs 380± 159, p=0.000). 4. Duration of morning stiffness in patients after BV therapy was significantly reduced compared with that before BV therapy(119.1± 112.6 min vs 59.0±89.7 min, p=0.009). 5. ESR and CRP were not significantly changed before and after BV therapy, suggesting BV itself could make inflammatory reaction as well as therapeutic effect. Conclusions BV therapy improved tender joint counts, swollen joint counts and duration of morning stiffness in this study, and further study is needed on long-term effect of BV therapy.

Angiotensin Concerting Enzyme 유전자의 유전적 다형성(Genetic Polymorphism)이 관상동맥 질환 위험인자에 따른 혈중 Plasminogen Activator Inhibitor-1의 농도에 미치는 영향

김덕경,김종원,권현절,류재춘,박승우,김준수,이상훈,홍경표,박정의,이원로 대한고혈압학회 1996 대한고혈압학회지 Vol.2 No.2