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재조합 균주 Escherichia coli가 생산하는 Bacillus stearothermophilus Exo-xylanase의 정제 및 특성
문애란,최용진 한국산업미생물학회 1992 한국미생물·생명공학회지 Vol.20 No.5
Bacillus stearothermophilus exo-xylanase 유전자 DNA가 삽입된 재조합 plasmid pMG1을 가지고 있는 E. coli JM109 exo-xylanase 생산 최적 배양 조건, 생산 효소의 정제 및 정제 효소의 특성 등을 조사연구하였다. 상기 재조합 E. coli 균주는 0.5% fructose, 0.5% yeast extract, 1.O% tryptone 및 1.O% sodium chloride가 함유된 배지에 서 약 10시간 배양했을 때 최대량의 효소를 생산하였으며 생산효소의 94%는 세포내에 존재하는 것으로 분석되었다. 생산 효소는 ammonium sulfate 분획, ion exchange chromatography및 gel filtration 등의 과정을 거쳐 단일 단백질로 정제하였으며 정제 효소는 pH 6.0과 45℃에서 가장 높은 효소 활성을 나타내었다. 또한 1 mM Ca^2+과 Co^2+ 이온의 첨가는 각각 약 25% 정도의 활성화 효과를 나타내는 반면, 본 효소의 ρNPX에 대한 K_m값은 2.75mM, pI값은 4.7, 그리고 분자량은 gel-filtration 법으로는 약 200,000 dal., SDS-polyacrylamide gel 전기 영동법으로는 약 66,000da1.으로 측정되어 세개의 동일한 subunit로 구성된 효소 단백질인 것으로 추정되었다. 본 정제 효소는 xylobiose, xylotriose 및 xylotetraose 등의 xylo-oligosaccharide를 효과적으로 분해함은 물론이고, 분해율은 낮으나 birchwood xylan, larchwood xylan 및 oatspelt xylan 등의 xylan에도 작용, xylose 생산을 확인함으로써 본 효소는 그 예가 극히 드문 bacterial exo-xylanase인 것으로 분류되었다. Exo-xylanase encoded by the xylA gene of Bacillus stearothermophillus was produced from Escherichia coli JM109 carrying a recombinant plasmid pMGl. Synthesis of the enzyme was observed to be cell-associated, and about 94% of the enzyme synthesized was located in the cytoplasmic region. The maximum production was attained when the E. coli strain was grown at 37℃ for 8 hours on the medium containing 0.5% fructose, 1.O% tryptone, 1.O% sodium chloride, and 0.5% yeast extract. The exo-xylanase was purified to homogeneity using a combination of salting out with ammonium sulfate, DEAE-Sepharose CL-6B ion exchange chromatography, Sephadex G-100 gel filtration, and Sephadex G-150 gel filtration. The purified enzyme was most active at pH 6.0 and 45℃ Ca^2+ and Co^2+ activated the exo-xylanase activity by about 20%, while Ag^2+, Fe^2+, Mg^2+ and Zn^2+ inhibited the enzyme activity by up to 60%. The K_m value on ρ-nitrophenyl-β-D-xylanopyranoside was 2.75 mM. The enzyme had a pI value of 4.7. The estimated molecular weight of the native protein was 200,000 dal. SDS-polyacrylamide gel electrophoresis analysis suggested that the native enzyme was a trimer composed of three identical 66,000 dal. olypeptides. The purified enzyme efficiently converted all the xylo-oligosaccharides tested to xylose. It was also confirmed that the enzyme split xylans in an exo-manner even though the degree of hydrolysis was fairly low. The xylanolytic enzyme was, therefore, classified to be one of the few bacterial exo-xylanases lacking transferase activity.
이원철,문애란 가톨릭대학산업의학센타 산업의학연구소 1996 韓國의 産業醫學 Vol.35 No.2
This study was conducted to evaluate possible correlation between mercury exposure and subjective symptoms among 201 mercury exposed workers and 132 controls. The mercury Exposed workers in this study have been working at 7 fluorescent lamp manufacturing factories. The Questionnaire were developed to evaluate subjective symptoms of the study subjects. Air mercury concentration and urinary mercury concentration were used as the index of mercury exposure. The mercury exposed work were divided by two group : one is below 100㎍/l of urinary mercury concentration and the other is over 100㎍/l of urinary mercury concentration and the other is over 100㎍/l of urinary mercury concentration. So, all the subjects were divided by 3 group : high concentration group, low concentration group and control group. Mean value of subjective symptom score of each group were composed and correlation coefficience among symptom score, air mercury concentration and urinary mercury concentration were estimated. The results were as follows; 1. Average mercury concentration in air showed 0.038㎎/㎥. Average mercury concentration in the low concentration group was 0.031㎎/㎥ and high concentration group was 0.099㎎/㎥. 2. Average mercury concentration in urine of workers was 55.06㎍/l, The urinary concentration of 21 Workers(10.5%) exceeded warning level(100㎍/l), and urinary concentration of 5 workers(2.5%) exceeded diagnostic criteria level(300㎍/l). Average mercury concentration in the low concentration group was 30.88㎍/l, in high concentration group was 246.20㎍/l. 3. There was significant difference of subjective symptom 12 items between mercury exposure group and non-exposure group. 4. There were no difference among subjective symptoms by work duration on the three group. 5. There was significant correlations among subjective symptoms and urinary mercury concentration on high concentration group, but low concentration group was not correlations. 6. There was significant correlations among subjective symptom 12 items and air mercury concentration on high concentration group and low concentration group.
산란계에서 퀴놀론계 약물투여후 혈장 및 계란내의 잔류함량 변화추이 조사
심애란 ( Ea Ran Sim ),김미희 ( Mi Hee Kim ),유은아 ( Eun Ah Yoo ),이윤정 ( Yun Jung Lee ),천순용 ( Soon Yong Chun ),문수평 ( Soo Pyeong Moon ),함유식 ( Yoo Sik Hahm ) 한국가축위생학회 2005 韓國家畜衛生學會誌 Vol.28 No.3
The purposes of this study were to evaluate the distribution of quinolone and to investigate the effects of quinolones (enrofloxacin, ciprofloxacin) in blood(plasma) and eggs of laying hens. Animals were fed quinolones which supplemented with 20, 50, 80㎎/㎏ of body weight. Blood and egg samples were collected after oral administration and analyzed for quinolones(enrofloxacin, ciprofloxacin) by HPLC. In laying hens, the residue period of enrofloxacin were longer than that of ciprofloxacin and the levels of residues were elavated by drug dosage.
Therapeutic Inhibitors against Mutated BRAF and MEK for the Treatment of Metastatic Melanoma
류순효,윤차경,문애란,Amanda Howland,Cheryl A. Armstrong,Peter I Song 전남대학교 의과학연구소 2017 전남의대학술지 Vol.53 No.3
Melanoma is one of the most aggressive cancers in the world and is responsible for the majority of skin cancer deaths. Recent advances in the field of immunotherapy using active, adoptive, and antigen-specific therapeutic approaches, have generated the expectation that these technologies have the potential to improve the treatment of advanced malignancies, including melanoma. Treatment options for metastatic melanoma patients have been dramatically improved by the FDA approval of new therapeutic agents including vemurafenib, dabrafenib, and sorafenib. These kinase inhibitors have the potential to work in tandem with MEK, PI3K/AKT, and mTOR to inhibit the activity of melanoma inducing BRAF mutations. This review summarizes the effects of the new therapeutic agents against melanoma and the underlying biology of these BRAF inhibitors.