• Journal of Pharmaceutical Investigation
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• 제22권2호
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• pp.115-124
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• 1992

The characteristics and stabilities of phosphatidylcholine liposomes covalently coupled with immunoglobulin fragments prepared by the REV method were investigated by the dynamic light scattering, absorbance and calcein release. Using a sulfhydryl-reactive phospholipid derivative of N-[4$({\rho}-maleimido-phenyl)$ butyl] phosphatidylethanolamine (MPB-PE), Fab' antibody fragments were covalently combined with preformed large unilamellar vesicles (LUV), Coupling ratio was $250\;{\mu}g$ of $Fab'/{\mu}mol$ of phospholipid in vesicles, From dynamic light scattering, it was found that the size of the vesicles increases as the ratio of cholesterol to lipid increases, but that apparently, the size of liposomes was not sensitive to the existence of Fab' fragments. Regardless of inserting Fab' fragments, the absorbance of liposomes decreased as the amounts of bile salt (BS) added. At very low BS concentrations, BS/lipid aggregates would be formed in the outer vesicles monolayer, while, at the high BS concentrations, mixed micelles would be preferred. The vesicles incorporated with Fab' fragments, however, are more resistant to the bile salts than the MPB-PE vesicle are. The absorbance of vacant liposomes and calcein release resulted in that the Fab' vesicles and MPB-PE vesicles by the REV method are very stable, but that those by the sonication method sufferred the significant change of turbidities.

• 한국환경보건학회지
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• 제23권3호
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• pp.91-101
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• 1997

Cycloheximide와 nalidixic acid를 처리한 배지에 Chlorella ellipsoidea를 배양하였을 때 mitochondria의 인지질 생합성과 그의 지방산 조성에 어떤 영향을 미치는 가를 분석하였다. 생장율과 total lipid 함량은 항생제 처리구에서는 대조구보다 낮게 나타났다. PC와 PI의 합성은 nalidixic acid 처리구에서는 억제되었고, PC, PE, PG 그리고 PI의 함량은 cycloheximide 처리구에서 억제되었다. 항생제 처리구에서 여러가지 인지질 형성에 이용된 주요 지방산은 배양 말기에 stearic acid, myristic acid, palmitic acid, oleic acid, linoleic acid 및 linolenic acid들 인것이 분석되었다.

• KSBB Journal
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• 제10권3호
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• pp.231-236
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• 1995

Saccharomyces sake Kyokai No.7을 이용한 고정화 균체 반응기에서 연속 알콜생산시에 발효배지 성분을 1/10배 ~3/10배 범위에서 희석하여 사용할 때 균체의 생산성과 고정화 beads의 물리적 안정성이 장기적으로 유지되었다. Egg albumin hydrolysate (0.5%)와 phosphatidylcholine(0.5%)을 첨가한 발효배지를 3/10배 희석하여 공급할 때 $1.1lhr^{-1}$ 희석 율에서 최대 알콜생산성인 $69 g/\ell$-hr 이 얻어졌으며, 이는 무첨가물의 2/1011H 희석 발효배지의 최대치인 $46 g/\ell$-hr 에 비하여 50% 증가한 것이다.

• BMB Reports
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• 제29권1호
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• pp.11-16
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• 1996

The present study showed that receptor-mediated activation of rabbit kidney proximal tubule cells by angiotensin II, the $Ca^{2+}$ ionophore A23187, or the protein kinase C activator phorbol myristate acetate (PMA) all stimulated phospholipase D (PLD). This was demonstrated by the increased formation of phosphatidic acid, and in the presence of 0.5% ethanol, phosphatidylethanol (PEt) accumulation. Angiotensin II leads to a rapid increase in phosphatidic acid and diacylglycerol, and phosphatidic acid formation preceeded the formation of diacylglycerol. This result suggests that some phosphatidic acid seems to be formed directly from phosphatidylcholine hydrolyzed by Pill. On the other hand, EGTA substantially attenuated angiotensin II and A23187-induced PEt formation, and when the cells were pretreated with verapamil angiotensin II-induced Pill activation was completely abolished. These results provide the evidence that calcium ion influx is essential for the agonist-induced Pill activation. In addition, staurosporine, an inhibitor of protein kinase C, strongly inhibited PMA-induced PEt formation, but was ineffective on angiotensin II-induced PEt accumulation. $GTP{\gamma}S$ also stimulates PEt formation in digitonin-permeabilized cells, but pretreatment of the cells with pertussis toxin failed to suppress angiotensin II-induced PEt formation. From these results, we conclude that in the rabbit kidney proximal tubule cells the mechanisms of angiotensin II- and PMA-induced Pill activation are different from each other and mediated via a pertussis toxin-insensitive trimeric G protein.

• 한국식품조리과학회지
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• 제27권3호
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• pp.51-57
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• 2011

Color, lipid and fatty acid composition, and tocopherols and polyphenols contents of perilla seed lipids in response to seed germination were studied. Perilla seeds were germinated at $30^{\circ}C$ in the dark for 12, 36, or 48 h, after which total lipids were extracted by the Folch method using chloroform and methanol (2:1, v/v). Seed germination resulted in a decrease in yellowness and greenness in perilla seed lipids, but there were no significant changes in composition of the lipids including major neutral lipids (>90%). Contents of phosphatidylcholine and phosphatidylethanolamine in the perilla seed lipids significantly increased in response to germination. Linolenic acid (>63%) was the most abundant fatty acid. Seed germination tended to decrease the relative content of linolenic acid and increase the contents of oleic and stearic acids. Contents of antioxidants, especially ${\alpha}$-tocopherol and polyphenols, increased in response to seed germination. As the germination period was extended, the antioxidant content increased. Therefore, increases in useful components, phosphatidylcholine, phosphatidylethanolamine, ${\alpha}$-tocopherol, and polyphenols contents by seed germination can contribute to the improvement of perilla seed utilization in food industry.

• 미생물학회지
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• 제40권3호
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• pp.211-216
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• 2004

세포외 phospholipase D (PLD)를 과량 생산하는 균주 JE-11을 토양으로부터 분리하였다. 16S rDNA에 의한 분석과 형태적, 생리적 특성을 조사한 결과 이 균은 Streptomyces somaliensis로 동정되었다. 선발한 S. somaliensis로 부터 PLD를 암호화하는 유전자(sspld) 분리하고 염기서열을 조사하였다. Open reading frame을 분석한 결과 33개의 아미노산으로 이루어진 분비 signal peptide와 505개의 아미노산으로 구성된 PLD단백질을 암호화하는 것으로 예상되었다. 또한, sspld의 염기 서열로부터 유추된 단백질 서열은 기존에 보고된 다른 Streptomyces PLD들과 70-88％의 서열 유사성을 보였다. 이 PLD는 96-98％(㏖/㏖)의 수율로서, Phosphatidylcholine을 glycerol과 serine을 기질로 하여 각각 phosphatidylglycerol 과phosphatidylserine으로 전환을 하였으나, 알코올 공여체인 inositol과 ethanolamine과는 반응하지 않았다.

• 대한화학회지
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• 제44권5호
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• pp.448-452
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• 2000

포스포리파제 D(PLD)는 인지질의 말단염기를 가수분해하여 phosphatidic acid(PA)와 염기로 분리시키는 효소이다. 본 연구는 쥐의 뇌에서 분리한 미토콘드리아 분획에서 PLD를 활성에 미치는 spermine의 영향을 조사하였다. 올레산 존재하에서 spermine이 PLD를 활성화시켰으며 $Ca^{2+}$, $Mg^{2+}$ 그리고 $Ba^{2+}$는 spermine의 영향을 더욱 강화시켜주는 것으로 나타났다. 다른 아민들은 별 효과가 없었으나 histamine경우 높은 농도에서 PLD를 활성화시켰다. Polylysine들도 PLD를 활성화시켰으며 그 길이가 길수옥 더욱 효과적으로 작용하였다. 기질 특이성에 있어서는 phosphatidylcholine(PC)과 phosphatidylethanolamine(PE) 사이에 큰 차이를 보이지는 않았다. 이 기질 특이성은 쥐 소장 미토콘드리아 PLD의 PE 특이성과 상이한 것으로 나타났다.

• Journal of Pharmaceutical Investigation
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• 제37권1호
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• pp.51-55
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• 2007

Solid lipid nanoparticles (SLNs) have been regarded to behave similar to the vegetable oil emulsions because emulsions of lipid melts are formed before lipid droplets being solidified to turn into SLNs. Compared to lipid emulsion, however, it has been more difficult to obtain stable SLNs and needs more extensive considerations on stabilizer and manufacturing process. In the present study, we tried to prepare phosphatidylcholine-based trymyristin (TM) SLNs using high pressure homogenization method and optimize the manufacturing variables such as homogenization pressure, number of homogenization cycles, cooling temperature, co-stabilizer and freeze-drying with cryoprotectants. Nano-sized TM particles could be Prepared using egg Phosphatidylcholine and pegylated phospholipids ($PEG_{2000}$PE) as stabilizers. Based on the optimization study, the dispersion was manufactured by homogenization under the pressure of 100 MPa for more than 5 cycles, and solidifying the intermediately formed lipid melt droplets by dipping in liquid nitrogen followed by thawing at room temperature. In addition, TM SLNs could be freeze-dried and then redispersed easily without significant particle size changes after freeze drying with 10% and 12.5% sucrose or trehalose. The TM SLNs established in this study can be used as delivery system for drugs and cosmetics.

• Journal of Dairy Science and Biotechnology
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• 제14권2호
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• pp.195-206
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• 1996

Cholesterol assimilated by Lactobacillus acidophilus ATCC 43121 was not metabolically degraded in that most of it was recovered with the cells. Cells grown in the presence of cholesterol micelles and bile salts were more resistant to Iysis by sonication than those grown in their absence, suggesting a possible alteration of cellular membranes. Cholesterol assimilation occurred during growth at pH 6.0, the amount of which was more than that by cells grown without pH control. Cholesterol assimilated by cells was recovered in the membrane fractions of cells both grown at pH 6.0 and without pH control. The effect of unsaturated fatty acids on cholesterol assimilation was not clear, since there was no significant (P> 0.05) difference in the amount taken up from micelles prepared using L-${\alpha}$-phosphatidylcholine, dioleoyl or L-${\alpha}$-phosphatidylcholine, distearoyl. Without Tween 80, little, if any, cell growth or cholesterol uptake was observed. In the presence of 0.05% Tween 80, cholesterol uptake increased dramatically as did growth. However, as the amount of Tween 80 increased beyond 0.05%, cholesterol uptake decreased while the amount of growth remained the same.

• Bulletin of the Korean Chemical Society
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• 제18권7호
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• pp.761-766
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• 1997

Phospholipase C from Clostridium perfringens is known to catalyze the hydrolysis of phospholipids in biological membranes. In this study, a simple and sensitive method for assaying phospholipase C was developed by using liposomes entrapping calcein as a fluorescent marker. Phospholipase C-induced lysis of liposomes was determined by measuring the fluorescence intensity of calcein released out from liposomes, Various liposomes with different compositions were prepared by reverse-phase evaporation method to investigate the effect of liposomal composition on the lytic activity of phospholipase C. The calcein-entrapping efficiency of liposomes was affected by the chain length of fatty acid in phosphatidylcholine constituting liposomes. The lytic activity of phospholipase C was the highest against liposomes prepared with eggPC. The lytic activity decreased with increasing chain length of fatty acid in phosphatidylcholine. Incorporation of cholesterol more than 20% into the liposomal bilayer inhibited the phospholipase C-induced lysis. The lysis of liposomes was more greatly increased by the addition of 10 mM of calcium. The lytic activity of phospholipase C was also affected by the surface charge of liposomes. Taken together, it was concluded that reverse-phase evaporation vesicles composed of dipalmitoylphosphatidylcholine and cholesterol in the molar ratio of 9 : 1 allowed to detect the lowest concentration of phospholipase C (0.10 μg/assay volume). This study suggested that the use of liposomes can provide a simple, sensitive and inexpensive method for assaying phospholipase C.