• Title/Summary/Keyword: P fraction

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Soluble Expression of the Fucosyltransferase Gene from Helicobacter pylori in Escherichia coli by Co-expression of Molecular Chaperones (샤페론단백질동시발현기술을이용하여 Helicobacter pylori 유래의 fucosyltransferase의수용성생산)

  • Lee, A Reum;Li, Ling;Shin, So-Yeon;Moon, Jin Seok;Eom, Hyun-Ju;Han, Nam Soo
    • Microbiology and Biotechnology Letters
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    • v.43 no.3
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    • pp.212-218
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    • 2015
  • Fucosyltransferases (FucTs) catalyze fucosyl transfer from guanosine-diphosphate fucose (GDP-β-L-fucose) to acceptor molecules to form fucosyloligosaccharides with α-glycosidic linkages. However, when FucT genes have been expressed in Escherichia coli, most cases have resulted in the production of inclusion bodies. In this study, to overcome this drawback, molecular chaperones were co-expressed with α1,2-fucosyltransferase (FucT2) in E. coli. For this, the pACYC184 vector, having genes for chaperones such as GroEL, GroES, DnaK, DnaJ, and GrpE, were transformed into E. coli BL21 (DE3) star harboring pHFucT2, including the FucT2 gene from Helicobacter pylori 26695. The results from SDS-PAGE showed that 5 chaperones were successfully expressed and the soluble fraction of FucT2 was also increased. HPLC analysis revealed that the coexpression of chaperone proteins resulted in a 5-fold increase in the total activity of fucosyltransferase in E. coli. In conclusion, the FucT2 expression system developed in this study can be used as a useful tool for the synthesis of fucosyloligosaccharides.

Effects of Barbiturates on the Fluidity of Phosphatidylethanolamine Model Membranes (Barbiturates가 소의 신선한 대뇌피질 Synaptosomal Plasma Membrane Vesicles로 부터 추출하여 제제한 Phosphatidylethanolamine 인공세포막의 유동성에 미치는 영향)

  • Yun, Il;Kim, Hyung-Il;Hwang, Tae-Ho;Kim, Jong-Ryol;Kim, In-Se;Chung, Yong-Za;Shin, Yong-Hee;Jung, Hyun-Ok;Kang, Jung-Sook
    • The Korean Journal of Pharmacology
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    • v.26 no.2
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    • pp.209-217
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    • 1990
  • Intramolecular excimer formation with 1,3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization with 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to evaluate the effects of barbiturates on the bulk fluidity of the model membranes of phosphatidylethanolamine fraction of synaptosomal plasma membrane vesicles (SPMVPE) isolated from bovine cerebral cortex. In the SPMVPE, barbiturates decreased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py and increased the fluorescence polarization (P), anisotropy (r), limiting anisotropy $(r_{8})$, order parameter (S) and rotational relaxation time $({\bar{P}})$ of DPH in a dose-dependent manner. The relative potencies of barbiturates to order the SPMVPE were in the order: pentobarbital > hexobarbital > amobarbital > phenobarbital. Hence, it is concluded that barbiturates have ordering effects on the SPMVPE. And the membrane-ordering potencies of barbiturates appear to be correlated with the potencies for enhancement of GABA-stimulated chloride influx and with the anesthetic effects of barbiturates.

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Standardization Studies for the Oriental Mineral Medicine (광물성 약재(광물약)의 표준화에 관한 연구)

  • Kim, Seon-Ok;Park, Maeng-Eon
    • Economic and Environmental Geology
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    • v.48 no.3
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    • pp.187-197
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    • 2015
  • Oriental mineral medicines are single or mixture of more than one mineral species or rock/fossil which are used to treat disease. Mineral medicines remove harmful or useless substances to decrease toxicity and secondary effects, and cause the manufacture of medical compounds with increased efficacy. The extraction test is an accepted in vitro system to predict the bioaccessibility of major and minor elements from mineral medicine. It incorporates gastrointerstinal tract parameters representative of a human body that including stomach and small intestinal pH which are the same as digestion condition. The bioaccessibility of a mineral medicine is the fraction that is soluble in the gastrointestinal environment and is available for absorption. Reaction path modeling in the human body can predict digestion with gastric fluid as well as absorption in the small intestine, existence in body fluids and reaction progress of the exhaust process according to pH conditions in body. Also reaction path modeling can predict bioavailability, which is equal to existence rate in the body and the form and amount of a medicine in the body after intake. The study results from predicating the existence form mineral medicines in the body, and proving the effective ingredient using bioaccessibitily and human risk assessment, suggest these that should be necessary data for new medicine development.

Consumption of Water-Soluble Egg Yolk Extract on Growth Rate, Changes in Blood Cholesterol Levels, and Immune Modulation in BALB/c Mice

  • Lee, Won-Young;Lee, Ran;Kim, Hee-Chan;Lee, Kyung-Hoon;Noh, Kyung Sook;Kim, Hyoun Wook;Kim, Ji-Hyuk;Ahn, Dong-Uk;Jang, In-Surk;Jang, Aera;Lee, Hoon-Taek;Song, Hyuk
    • Food Science of Animal Resources
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    • v.33 no.5
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    • pp.587-594
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    • 2013
  • Egg consumption has been limited to avoid cardiovascular diseases such as atherosclerosis and hyperlipidemia, because the yolk contains high levels of cholesterol. This study was conducted to evaluate the effect of the water-soluble component of egg-yolk on the growth efficiency, immune modulation, and changes in serum lipid levels in BALB/c mice. A total 5 wk old 120 BALB/c male mice were divided into 4 groups and were fed 0, 2, 10, and 20 mg/d water-soluble egg yolk extract (WSEYE) for 5 wk. Water-soluble egg yolk extract (WSEYE) uptake resulted in a significant reduction in daily weight gain and feed efficiency rate (FER). The mouse groups treated with 2 and 20 mg/d WSEYE showed a significant increase in populations of monocytes at the third wk and B-lymphocyte activity at the fifth wk. In addition, WSEYE uptake did not influence serum immunoglobulin E levels. In serum lipid-profile studies, treatment of WSEYE did not alter total cholesterol and low-density lipoprotein levels; however, blood triglyceride levels were significantly diminished in mice treated with 2 mg/d at the third wk (p<0.05), and the level of high-density lipoprotein was significantly increased in the mice group treated with 2 and 10 mg/d WSEYE after 5 wk (p<0.05). Taken together, the data demonstrate the beneficial effects of WSEYE in the diet on immune modulation and serum lipid profiles in mouse models; therefore, this study suggests that ingestion of water-soluble fraction of egg yolk might not be related to the increased risk of heart disease, but can be an excellent candidate for maintaining health.

Biochemical properties of a purified protein in cystic quid of Taenia solium metacestodes (유조낭고충 낭액에서 친화성 크로마토그래피로 분리한 항원 단백질의 생화학적 성상)

  • Cho, Seung-Yull;Kim, Suk-Il;Kang, Shin-Yong;Kong, Yoon
    • Parasites, Hosts and Diseases
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    • v.26 no.2
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    • pp.87-94
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    • 1988
  • By affinity chromatography using a monoclonal antibody as ligand, Kim et at. (1986) purified a protein fraction in cystic fluid of Taenia solium metacestodes (CF) In this study, the biochemical properties of the purified protein were characterized. Discontinuous-polyacrylamide gel electrophoresis (disc-PAGE) of the protein at 4.5∼10% separating gel concentration showed its molecular weight (MW) to be 150 kilodalton (kDa) in non·denatured state, while denaturing sodium dodecyl suifate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that it was composed of 3 different subunits with respective fnw of 15, 10 and 7 kDa. Subunit of 7 kDa was shown to be linked to other subunits by disulade bonds. Isoelectric point of the protein was pH 6.8. The protein was relatively heat-stable for immunologic analysis. These properties indicated that the protein, comprising about 70% of total content in CF, had similar biochemical characters with antigen B of Oriol et at.(1971) in hydatid cyst quid (HF).

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Isolation and Characterization of MMP-1 Inhibitor Peptide from Crataegus pinnatifida Bunge in Fibroblast Cell Line HS68 Cells (아가위(Crataegus pinnatifida Bunge)로 부터 HS 68세포의 MMP-1에 대한 저해활성 물질의 분리)

  • Lee, Se-Young;Chun, Hyug;Cho, Hong-Yun;An, Jeung-Hee
    • Applied Biological Chemistry
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    • v.46 no.1
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    • pp.60-65
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    • 2003
  • MMP-1 inhibitory compounds were isolated from 120 Korean traditional edible plants. UP- 1 activity significantly increased linearly with increasing UVB dose in normal human foreskin fibroblast HS68 cell, showing maximum activity at approximately 35 $mJ/cm^2$, whereas in HaCaT cell, normal human keratinocyte, no increase was observed. Maximum secretion of MMP-1 after UVB treatment occurred around 36-48 k after treatment. MMP-1 inhibitory compound isolated from cold-water fraction of Cataegus pinnatifida Bunge showed the mort potent activity. The MMP-1 inhibitory compound was deduced as a peptide based on the fact that pronase digestion decreased the activity whereas periodate oxidation did not. The most potent UP- 1-inhibitory protein, CP-2Va-2, showing an activity of 88.5% against MMP-1, was isolated through sequential column chromatography on DEAE-Toyopearl 650C, Butyl-Toyopearl 650M, and Bio-Gel P-30. Molecular weight of CP-2Va-2 determined through high performance liquid chromatography and SDS PACE was 19 and 20 kDa. respectively, signifying a monomeric structure.

Comparison of Regional Myocardial Perfusion, Left Ventricular Volume and Ejection Fraction between 8- and 16-Frame Gated SPECT (8-단위영상과 16-단위영상으로 얻은 게이트 심근관류 스펙트에서 국소 심근관류, 좌심실 용적과 구혈률의 비교)

  • Lee, Sang-Woo;Lee, Jae-Tae;Bae, Jin-Ho;Jeong, Shin-Young;Chun, Kyung-Ah;Ha, Jeoung-Hee;Ahn, Byeong-Cheol;Lee, Kyu-Bo
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.5
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    • pp.269-277
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    • 2003
  • 목적: 게이트 심근관류 스펙트는 심근의 관류와 좌심실 기능을 동시에 평가할 수 있는 유용한 방법이다. 현재, 게이트 영상 획득시 8 단위영상이 널리 사용되지만, 아직 가장 적절한 단위영상의 수에 대해서는 논의의 여지가 있다. 본 연구는 동일한 환자에서 8 단위영상과 16 단위영상으로 게이트 심근관류 스펙트를 2회 연속적으로 시행하여 국소적 심근관류, 좌심실 용적과 구혈률등을 비교하였다. 대상 및 방법: 42명(평균 연령: 55세, 남: 녀=26: 16)의 관상동맥 질환이 의심되거나 알려진 환자를 대상으로 안정시 Tc-99m MIBI를 이용한 게이트 스펙트를 8 단위영상과 16 단위영상으로 2회 연속하여 시행하였다. 좌심실의 확장기말 용적, 수축기말 용적과 구혈률을 각각 얻었으며 확장기말의 단층영상을 18 분절로 나누고 각 분절마다 심근의 관류정도를 4등급으로 나누어 국소적 심근관류를 평가하였다. 결과: 8 단위영상과 16 단위영상 게이트 스펙트에서 국소적 심근관류의 일치도는 85.7% (tau-b=0.786, p<0.001)로 우수하였다. 두 방법간에 1등급의 차이는 100 분절(13.2%), 2등급 차이는 8 분절(1.1 %)이 있었고 3등급 차이는 없었다. 좌심실의 확장기말 용적, 수축기말 용적, 구혈률은 두 방법간에 높은 상관성을 보였으며(r=0.993, 0.989, 0.935), 좌심실 구혈률은 8 단위영상을 사용할 때 평균 1.8% 낮게 측정되어 통계적인 유의성은 있었으나(p<0.05), 그 차이는 크지 않았다. 결론: 8 단위영상과 16 단위영상을 사용한 게이트 심근관류 스펙트에서 국소적 심근관류의 일치도는 우수하였으며, 좌심실의 기능적 계수의 차이도 크지 않았다. 비록 높은 단위영상을 사용했을 때 더 정확하고 자세한 정보를 얻을 수 있다 하더라도, 구혈률과 용적 등을 얻는 일상적인 임상검사에는 8 단위영상 또한 그 역할을 충분히 할 수 있을 것으로 사료된다.

Effects of Hot Water Extracts from Lentinus edodes on Hepatic Functional Enzyme Activities in the Rat Fed Butter Yellow(p-Dimethylaminoazobenzene) (표고버섯 열수 추출물이 발암원을 급여한 흰쥐의 간 기능 관련 효소활성에 미치는 영향)

  • 최미연;정수자;임상선
    • The Korean Journal of Food And Nutrition
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    • v.11 no.1
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    • pp.114-122
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    • 1998
  • This study was designed to observe the effect of hot water soluble polysaccharides extract(PS) from Lentinus edodes on the enzyme activities related with hepatic function and peroxidation in the rats fed better yellow. The four groups of male SD rats were fed with the diets contained 15% casein(basal diet; NO group), added butter yellow(BO group) or /and PS(NP, BP group) for 6 weeks. The activities of ${\gamma}$-GTP and GPT in BP were significantly lower compared with BO. The activities of glutathione peroxidase, catalase and lactate dehydrogenase were not significantly different between NP and NO, while those activities were significantly lower value in BP than BO. The activities of glutathione S-transferase of the microsomal and cytosol fractions were significantly lower in BP than in BO. The contents of glutathione and malondialdehyde in the liver were considerably low value in BP. In a view of these results the PS of Lentinus edodes prevents the lipid peroxidation and diminishes the liver toxicity caused with better yellow. The superoxide dismutase activity in cytosolic fraction of liver was not found any effect in all groups. But hepatic function enzyme activities such as catalase and glutathione peroxidase, LDH activities were remarkably decreased in the groups 2(basal diet + PS) and the ${\gamma}$-GTP, GOT and GPT activities, too. In liver, the contents of glutathione decreased by PS supplementation but HDL-cholesterol and total cholesterol ratio in plasma decreased at the groups 3, 4. The ${\gamma}$-GTP, GOT and GPT in plasma were remarkably higher in the rats fed the p-DAB than the control group, too. But above enzyme activities significantly decreased in the groups fed PS.

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Protective Effects of Sasa Borealis Leaves Extract on High Glucose-Induced Oxidative Stress in Human Umbilical Vein Endothelial Cells (혈관내피세포에서 고농도 포도당으로 유도된 산화스트레스에 대한 조릿대잎 추출물의 보호효과)

  • Hwang, Ji-Young;Han, Ji-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.12
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    • pp.1753-1760
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    • 2010
  • This study was designed to investigate the protective effects of Sasa borealis leaves on high glucose-induced oxidative stress in human umbilical vein endothelial cells (HUVECs). Freeze-dried Sasa borealis leaves were extracted with 70% methanol and followed by a sequential fractionation with dicholoromethan, ethyl acetate, butanol and water. The ethyl acetate fraction from Sasa borealis leaves extract (ESLE) was used in this study because it possessed the strongest antioxidant activity among the various solvent fractions. Exposure of HUVECs to 30 mM high glucose for 48 hr resulted in a significant (p<0.05) decrease in cell viability, glutathion (GSH) concentration, activities of antioxidant enzymes including superoxide dimutase (SOD), glutathion peroxidase (GSH-px) and catalase, and a significant (p<0.05) increase in intracellular ROS and lipid peroxidation formation in comparison to the cells treated with 5.5 mM glucose. ESLE treatment decreased intracellular ROS and lipid peroxidation formation and increased cell viability, GSH concentration and expressions of SOD and catalase in HUVECs. These results suggest that ESLE may be able to protect HUVECs from high glucose-induced oxidative stress, partially through the antioxidative defense systems.

Antimicrobial Activity of Gluten Hydrolysate with Asp. saitoi Protease (밀 단백 효소 가수분해물의 항균활성)

  • Lee, Sang-Duk;Joo, Jeong-Hyeon;Lee, Gyu-Hee;Lee, K.T.;Oh, Man-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.5
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    • pp.745-751
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    • 2003
  • This study was carried out to investigate whether peptide produced from wheat protein by enzyme hydrolysis can be used as a natural antimicrobial agent. Antimicrobial peptide was obtained from wheat protein hydrolyzed by 7 of pretense. The produced antimicrobial peptide was purified through ultrafiltration, membrane filtration and HPLC and molecular weight and amino acid sequence of the purified antimicrobial peptide were determined. Among hydrolysate produced from wheat protein by 7 of protease, antimicrobial activity was observed for the peptide obtained from Asp. saito protease. The Asp. saito protease did produce antimicrobial hydrolysate showing the highest antimicrobial activity at reaction condition of 37$^{\circ}C$ and pH 6.0, but not at reaction condition above 5$0^{\circ}C$. Wheat protein hydrolysate was fractionated by membrane filtration and showed antimicrobial activity between molecular weight 1,000~3,000. The antimicrobial activity fraction obtained by membrane filtration was separated through HPLC and showed antimicrobial activity in the peak of retention time 31.1~31.8 min. We could convince this hydrolysate as heat-stable peptide since antimicrobial activity was maintained after treated with heat for 15 min at 121$^{\circ}C$. Molecular weight of antimicrobial peptide identified by MALDI-mass was 1,633. Amino acid sequence of antimicrobial peptide was cysteine, glycine, prolin, prolin, prolin, valine, valine, alanine, alanine and arginine.