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김영식,이미연,박영목 ( Yeong Shik Kim,Mi Youn Lee,Young Mok Park ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.2
Chitinase (EC 3.2.1.14) was purified from the mature tissue of green onion by ammonium sulfate precipitation followed by affinity chromatography on a regenerated chitin and a Sephacryl HR-100. The purified enzyme gave a single band on polyacrylamide slab gel electrophoresis and its molecular weight was determined to be 35,000 Da using SDS-PAGE and HPLC-GPC. The isoelectric points of the enzyme were 3.5 and 7.2. The purified enzyme was stable on incubation at 50℃ for up to 20 min. Most of metal ions did not affect the activity significantly except that Ag^+ and Hg^(2+) ions inhibited the enzyme activity at 10 mM concentration. HPLC analysis of the initial products from the digestion of colloidal chitin and chitooligosaccharides indicated that chitinase was endolytic in action, yielding oligomers from the dimer to higher oligosaccharides.
형광기 - 기질 결합체를 이용해 폴리아크릴아미드겔 위에서 chitinase 활성의 검정
김영식,이경복,Robert J . Linhardt ( Yeong Shik Kim,Kyung Bok Lee,Robert J . Linhardt ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.5
Gradient polyacrylamide gel electrophoresis was used to analyze the products formed by chitinases acting on N-acetylchitohexaose-fluorescent conjugate. N-Acetylchitooligosaccharides were conjugated to 7-amino-1,3-naphthalene disulfonic acid by reductive amination. Each oligosaccharide-fluorescent conjugate was purified by preparative gradient polyacrylamide gel electrophoresis, semi-dry electrotransfer to a positively-charged nylon membrane and recovered by washing the membrane with salt solution. N-Acetylchitohexaose-fluorescent conjugate and chitohexaose were exhaustively treated with three kinds of chitinases from Serratia marcescens, Streptomyces griseus, and green onin (Allium fistulosum L.). The bands were visualized under long wavelength of UV light. Analysis of reaction products provided the information on the action of chitinase action from different sources.
김영식,노지은,안형수,Kim, Yeong-Shik,Roh, Ji-Eun,Ann, Hyung-Soo 한국생약학회 1993 생약학회지 Vol.24 No.2
Polysaccharide from Sanguisorba officinalis was separated and fractionated using DEAE-Sephadex ion-exchange chromatography and Sephacry HR-200 gel filtration chromatography. One of the fraction(Fr. II) was sulfated and its anticoagulant activity was tested in vitro. Sulfation could increase the clotting time 50 times compared to unsulfated one. Fr. II was hydrolyzed and its composition was analyzed by conjugation with 7-amino-1, 3-naphthalene disulfonic acid using HPLC and electrophoresis. Arabinose and galactose were mainly composed at the ratio of 4 : 1. In addition, xylose and rhamnose were also found.
김영식(Yeong Shik Kim),노지은(Ji Eun Roh),안형수(Hyung Soo Ahn),박호군(Ho Koon Park) 대한약학회 1992 약학회지 Vol.36 No.4
Anticoagulant activities were tested for the fifteen kinds of medicinal plants by measuring activated partial thromboplastin time (aPTT). Of them five kinds of species (Artemisia princeps, Sanguisorba officinalis, Artemisia apiacea, Eclipa alba, Schizonepeta tenuifolia) were selected and fractionated for the preparation of acidic polysaccharides. They were extracted with water by refluxing and the extracts were precipitated with ethanol. The precipitates were separated based on charge using a DEAE-Sephadex. The low salt and high salt fractions were sulfated with anhydrous pyridine and chlorosulfonic acid complex. In vitro anticoagulant activities of sulfated polysaccharides were tested by measuring aPTT, prothrombin time (PT), and factor Xa clotting time using normal human plasma. No relationship was found between the amount of uronic acids and anticoagulant activities, but the sulfated ones show the increase of activities. In vivo anticoagulant properties of the sulfated polysaccharide from Artemisia apiacea were also tested by the intraveneous administration of three different doses (3, 5 and 10mg/kg) to rats. APTT and PT were increased significantly and the action of factor Xa and thrombin mediated through antithrombin III were inhibited slightly.
흰쥐의 일차배양 간세포에서 Daidzin, Daidzein, Genistein 및 Puerarin의 간 보호 활성 평가
박진구,천호준,김영식,강삼식,최재수,이선미,Park, Jin-Goo,Cheon, Ho-Joon,Kim, Yeong-Shik,Kang, Sam-Sik,Choi, Jae-Sue,Lee, Sun-Mee 대한약학회 2007 약학회지 Vol.51 No.2
The aim of this study was to investigate the protective activities of daidzin, daidzein, genistein or puerarin, active isoflavonoids of Puerariae Radix, on the hepatocyte injury induced by carbon tetrachloride (CCl$_4$, 10 mM), tert-butyl hydroperoxide (TBH, 0.5 mM) and D-galactosamine (GalN, 30 mM). Primary cultures of rat hepatocytes (18 hr cultured) were treated with CCl$_4$, TBH or GalN and various concentrations (0.1, 1, 10 and 100 ${\mu}$M) of daidzin, daidzein, genistein or puerarin. CCl$_4$ significantly increased the levels of lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The increase in LDH level was attenuated by daidzein, genistein and puerarin. Puerarin also inhibited the increase in AST level induced by CCl$_4$. The increases in LDH and ALT levels induced by TBH were significantly attenuated by daidzin and genistein treatments. GalN markedly increased the levels of LDH, ALT and AST These increases were significantly attenuated by daidzein. Daidzin also inhibited the increases in LDH and AST levels induced by GalN. The increases in LDH and ALT levels were attenuated by genistein and puerarin, respectively. These results suggest that daidzin and daidzein possess hepatoprotective activities.