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Escherichia coli(M15)을 이용한 serotonin N-acetyltransferase 유전자의 발현
김용만,정미영,윤경식,진병관,백행운,조용호,백형환 慶熙大學校 1997 論文集 Vol.26 No.-
AA-NAT cDNA was obtained by RT-PCR technique from total RNA of rat sacrified at night(02:00am). pCRNAT was cloned using the pCRII vector with insertion of AA-NAT cDNA(about 1.4 kb) at EcoRI site. For the expression of the gene, pQECNAT was subcloned, in which the coding region of AA-NAT was inserted into expression vector pQE30 at BamHI and HindIII sites. According to the experimental results, Escherichia coli strain M15, transformed by the expession vector pQECNAT, was selected as a host to express AA-NAT gene with the induction of isopropyl β-D-thiogalactoside(IPTG). Optimal condition for the expression of AA-NAT gene was achieve from the experimental results, showing that the expression of the protein was inducible much 19.800 ± 2,200 dpm per ml of culture volume in 4 hours at the concentration of 2 mM IPTG. Partial purification through the affinity column(Ni-NTA agarose) binding to the continuous 6 histidine residues of protein resulted in 5 times more increase in the specific activity of AA-NAT than that of the homogenate of bacterial pellet. These experimental results will provide basic data in further study for the enzymatic kinetics and antibody production of AA-NAT.
망막아세포종 세포주(Y-79)의 Arylalkylamine N-acetyltransferase 활성에 미치는 Dopamine의 영향
김석민,정미영,윤경식,진병관,백행운,백형환,조용호 慶熙大學校 1997 論文集 Vol.26 No.-
The arylalkylamine N-acetyltransferase(AA-NAT, EC 2.3.1.87) in vertebral pineal gland and retina is rate-limiting enzyme in the pathway of biosynthesis of melatonin. AA-NAT activities are remarkably increased during night with 10-150 folds. Neurohormone melatonin is thought to play roles in a broad range of physiological functions in human, primarily through effects on the biological clock in the suprachiasmatic nucleus of the hypothalamus, This rhythm reflects large changes in the activities of AA-NAT. So the important regulatory role of AA-NAT has made it central interest in research on melatonin biochemistry and neurochemical signal transduction. The neurotransmitter that regulates the AA-NAT activities is norepinephrine, which acts through a cyclic AMP mechanism. The effect of dopamine on AA-NAT in human retinoblastoma Y-79 cells was studied. the obtained results are as follows; AA-NAT activities in Y-79 cells were increased by cyclic AMP, and dopamine inhibited the AA-NAT activities stimulated by forskolin. The pattern of inhibition of dopamine on the AA-NAT activities in Y-79 cells was time and concentration dependent. The results suggest that dopamine participates in the regulation of the AA-NAT activities through the inhibitory mechanism in the human pineal and retinal tissues.
Baik, Hyung-Hwan,Cho, Yong-Ho 慶熙大學校 1992 論文集 Vol.21 No.-
소의 망막 색소상피세포를 분별원침법으로 수용성분획과 사립체 분획을 분리하여 세포내에 함유되어 있는 aspartate aminotransferase isozyme 의 활성도를 측정하고 두 분획내에 존재하는 효소를 부분정제하여 생활학적 성상을 관찰하였다. 또한 두 분획내에 존재하는 aspartate aminotransferase isozyme의 비활성도에 미치는 금속이온, 온도, pH, 효소 단백질량과 기질인 L-aspartate의 영향을 관찰한 바 다음과 같은 흥미있는 지견을 얻었다.
백서 신장 조직내 Na^+-K^+ ATPase에 대한 Furosemide의 간접적 억제 기전
백행운,윤경식,우태욱,김재욱,이재범,백형환,조용호 慶熙大學校 大學院 1994 高凰論集 Vol.15 No.-
It is presented that the subcellular localization and distribution of the specific activity of Na^+-K^+ ATPase in whole homogenate, the soluble fraction and mitochondrial fraction of the renal tissues in normal adult rat isolated by differential centrifugal method were studied. The effect of furosemide compound on their enzymatic activity was evaluated by the Fiske SubbaRaw method. The specific activity of Na^+-K^+ ATPase of rat renal tissues was found to be about 92.2 umol/g protein/hr. The specific activity of microsomal fraction of rat renal tissue is showed approximately 1.5-fold higher than the value in the other fraction. The effect of furosemide was found to decrease proportionally the specific activity of Na^+-K^+ ATPase in rat renal tissue as the increment of the concentration of furosemide to luM in the microsomal fraction. The effect of the other diuretics is showed the same inhibition pattern. When the incubation time is increased from 15min to 60min in the same drug treated Na^+-K^+ ATPase, specific activity of Na^+-K^+ ATPase is increase but the inhibition rate of the enzyme is decreased. In view of the above finding, it is clear that the furosemide, well known as diuretic agent, selectively inhibits the activity of the microsomal Na^+-K^+ ATPase in accordance with dose-dependent effect. The mechanism of action appears to be due to indirect inhibition. Indirect inhibition means that the transport protein of cellularion in membrane is inhibited by the furosemide and so indirectly activity of Na^+-K^+ ATPase is inhibited by that. Consequently, the effect of the furosemide on the rat renal tissue Na^+-K^+ ATPase is non-specific inhibition by indirect inhibition of transport protein.
Baik, Haing-Woon,Na, In-Gug,Baik, Hyung-Hwan,Cho, Yong-Ho 慶熙大學校 1989 論文集 Vol.18 No.-
흰 생쥐의 뇌조직으로부터 분별원심법으로 분리한 supernatant 분획과 mitochondrial 분획내 Ca^(2+)-ATPase의 비활성도와 세포내 국재소를 정하였고, 방사선 조사가 이 효소의 비활성도에 미치는 영향을 관찰한 결과 다음의 성적을 얻었다. 1. 흰 생쥐의 뇌조직으로부터 분리한 supernatant fraction내에 함유되어 있는 Ca^(2+)-ATPase의 활성도는 92.6 unit였고, mitochondrial fraction의 것으로 504.2 unit였다. 2. 흰 생쥐 뇌조직내의 세포질에 용해되어서 부유상태로 있는 supernatant Ca^(2+)-ATPase에는 총활성도의 15.5 퍼센트, mitochondrial fraction에는 84.4 퍼센트가 분산되어 있었다. 따라서 흰 생쥐의 뇌조직내에 존재하는 Ca^(2+)-ATPase는 주로 mitochondrial 분획내에 존재한다. 3. 뇌조직내 supernatant Ca^(2+)-ATPase 비활성도에 미치는 방사선 조사의 억제적 영향은 방사선량을 증가할수록 비례적으로 상승하였다. 즉 dose-dependent 관계가 있었다. 4. Supernatant Ca^(2+)-ATPase 비활성도를 50 퍼센트로 억제하는 방사선 조사량(I50-치)은 약 2870 Rad이었고, mitochondrial Ca^(2+)-ATPase는 2584 Rad였다. 5. Supernatant와 mitochondrial Ca^(2+)-ATPase 비활성도에 미치는 방사선 조사의 억제적 영향은 통계학적으로 매우 높은 유의성을 보였다. 이상과 같은 실험결과로 보면, 휜 생쥐 뇌조직내의 supernatant 분획과 mitochondrial 분획내에 존재하는 Ca^(2+)-ATPase 비활성도에 미치는 방사선 조사의 억제효과에 대한 기전은 방사서이 주로 효소분자 중의 효소단백 부분을 파괴함으로써 나타나는 것이라 시사된다. 그러나 방사선 조사가 흰 생쥐 뇌조직내 존재하는 Ca^(2+)-ATPase 비활성도에 미치는 억제효과와, 뇌종양 치료에 사용되고 있는 방사선 조사량과의 관련은 앞으로 더욱 연구가 진행되어야 하리라고 본다. An Experimental Study on the Inhibitory Effect of Irradiation upon Ca^(2+) - Mg^(2+) - activated Adenosine Triphosphatase Activity In Brain Tissues of Albino Mouse.
부착규조 군집과 유기오탁지수를 이용한 오산천의 수질평가
김백호(Baik-Ho Kim),최환석(Hwan-Seok Choi),김미연(Mi-Yeon Kim),유형빈(Hyung-Bin Yoo) 대한환경위생공학회 2004 대한환경위생공학회지 Vol.19 No.2
To investigate the epilithic diatom community and water quality of the Osan stream, water samples were collected from the eight stations from April to September 2003. Sampling was two times before and after heavy rain. Total 52 diatom were identified and divided into 12 saproxenosus taxa, 6 saprophilous taxa and 34 indifferent taxa, respectively. The DAIpo values higher after heavy rain than before that. According to tolerance degree to the organic water pollution, all sampling stations ranged from α-oligosaprobic to α-mesosaprobic. Thus, the result indicates that the water quality of Osan stream is gradually improved by heavy rain.
Purification of the Coolant for Hot Roller by Superconducting Magnetic Separation
Tae-Hyung Kim,Dong-Woo Ha,Jun-Mo Kwon,Myung-Hwan Sohn,Seung-Kyu Baik,Sang-Soo Oh,Rock-Kil Ko,Ho-Sup Kim,Young-Hun Kim,Seong-Kuk Park IEEE 2010 IEEE transactions on applied superconductivity Vol.20 No.3
<P>It is important to recycle industrial wastewater in the aspect of energy saving and minimizing the environment damage. Steel manufacture factories make an effort to purify and recycle wastewater from the manufacturing processes. A steel manufactory in Korea, POSCO (POhang iron and Steel COmpany) has also concerned about the treatment of wastewater for long time. The coolant of hot roller at steel making factory includes suspended solid (SS) like steel fines, iron oxide and emulsified oil. A laboratory-scale conduction cooled high temperature superconducting (HTS) magnet was used for high gradient magnetic separation (HGMS) systems. It consists of eight double-pancake sub-coils, two HTS current leads, a GM cryo-cooler and a cryostat without a radiation shield vessel. Sub coils were wound with 1 G HTS tapes of Sumitomo Electric Industries by wet-winding method. A Cryo-cooled high temperature superconducting magnet system with 70 mm room temperature bore and 250 mm of height was prepared. This magnet was used for batch type HGMS system. Superconducting HGMS system that had a purpose to purify the wastewater was assembled. Various magnetic seeding reactions were investigated to increase the reactivity of coagulation.</P>