豚鼠心脏CDP-乙醇胺磷酸转移酶的动力学研究

为探讨不同的二价基甘油对三种乙醇胺甘油磷脂生物合成能力的影响,通过对豚鼠乙醇胺磷酸转移酶动力学研究,发现磷脂酰乙醇胺的合成可被1-烷基-2-脂酰甘油和1-烯醚基-2-脂酰甘油抑制,而缩醛磷脂酰乙醇胺的生成不受1,2-二脂酰甘油影响,并提示不同的二价基甘油对乙醇胺磷酸转移酶的抑制作用呈非竞争性抑制,此有利于对三种乙醇胺磷脂酰甘油生物合成的相互协调作用。     

干旱条件下冬小麦幼苗根细胞膜脂组成的变化

干旱条件下小麦幼苗根膜脂总脂肪酸含量、磷脂含量及总脂肪酸双键指数均下降,而游离甾醇含量却明显增加,结果导致游离甾醇／磷脂比率上升。用薄层层析法测得小麦根细胞磷脂主要由磷脂酰胆碱（ＰＣ）、磷脂酰乙醇胺（ＰＥ）、磷脂酰肌醇（ＰＩ）及磷脂酸（ＰＡ）组成。干旱降低了各种磷脂的含量,但不改变其相对配比。文中讨论了膜脂代谢变化与植物抗旱性的关系     

干旱条件下冬小麦幼苗根细胞膜脂组成的变化

干旱条件下小麦幼苗根膜脂总脂肪酸含量、磷脂含量及总脂肪酸双键指数均下降,而游离甾醇含量却明显增加,结果导致游离甾醇／磷脂比率上升。用薄层层析法测得小麦根细胞磷脂主要由磷脂酰胆碱(PC)、磷脂酰乙醇胺(PE)、磷脂酰肌醇(PI)及磷脂酸(PA)组成。干旱降低了各种磷脂的含量,但不改变其相对配比。文中讨论了膜脂代谢变化与植物抗旱性的关系。     

豚鼠器官中缩醛磷脂公布的研究

豚鼠器官中缩醛磷脂公布的研究吕灿群，蔡镇潮（皖南医学院生物化学教研室，芜湖２４１００１）（加拿大玛尼托巴大学医学院生化与分子生物学系）关键词缩醛磷脂；缩醛磷脂酰乙醇胺；薄层层析；豚鼠缩醛磷脂（Ｐｌａｓｍａｌｏｇｅｎ）是存在于哺乳动物组织内含有烯醚键的...     

《要吃，就吃好的》———动物磷脂与植物磷脂的区别

《要吃,就吃好的》———动物磷脂与植物磷脂的区别磷脂（ｐｈｏｓｐｈｏｌｉｐｉｄ,ＰＬ）是一种脂类。脂类是不溶于水而溶于有机溶剂的天然产物。磷脂类主要包括：１．磷脂酰胆硷;２．磷脂酰乙醇胺;３．神经磷脂;４．缩醛磷脂。现在市面上出售的磷脂绝大多数是植物...     

去细胞外钙对培养鸡胚肌管磷脂酰肌醇水解的影响

本工作研究了去细胞外钙对取自９天来亨鸡胚的培养肌管磷脂酰肌醇水解的影响,８０ｍｍｏｌ／ＬＫ＾＋暴露可以显著和蔼同磷脂酰肌醇的水解。在暴露于无钙任氏液的肌管中,磷脂酰肌醇的水解随时间延长而呈指数递减。时间常数约为２６分钟。在胞外无钙时,８０ｍｍｏｌ／ＬＫ引起的磷胆酰肌醇水解与对照相比略有下降,结果表明,高钾暴露能够增强培养肌管的磷脂酰醇的水解;但与成熟的肌纤维不同,细胞外钙是必须的。     

酿酒酵母酚类抑制物耐受性脂质组学研究

通过脂质组学分析方法从细胞膜磷脂分布方面探究适应进化酿酒酵母酚酸耐受性机制。主要利用高效液相色谱-质谱(LC-MS)对酚酸胁迫下适应进化菌株和原始菌株脂质成分检测并进行统计学比较分析。检测出565种脂质代谢物,包含细胞膜磷脂185种。相比初始菌株,适应进化菌株细胞膜中磷脂酰胆碱(PC)、磷脂酰乙醇胺(PE)和磷脂酰肌醇(PI)类磷脂分子相对含量增加,含有长链(C32-C36)和双不饱和脂酰链的磷脂分子含量增加。统计学分析表明显著性差异磷脂分子主要为含有长链不饱和脂酰链的PC和PE类磷脂分子。推测适应进化菌株通过膜磷脂重塑提高细胞膜完整性,对酚类抑制物起到选择性屏障作用,从而保持细胞活性。     

冬虫夏草及杜仲磷脂成分的研究

本文对不同产地的冬虫夏草及杜仲磷脂成分进行了分析研究。以钼蓝比色法测定了它们的总磷脂含量,采用薄层色谱扫描和吸光度比例系数校正法测定了其磷脂组成及相对百分含量。冬虫夏草约含8种磷脂组分,主要成分为磷脂酰胆碱、磷脂酰叽醇、磷脂酰丝氨酸、磷脂酰乙醇胺和磷脂酸。杜仲约含6种磷脂组分,其中以溶血磷脂酰胆碱和磷脂酰胆碱为主。     

我国健康成人红细胞膜脂类的脂酸组成

 本文采用双向簿层层析分离红细胞膜脂类,继以毛细管气相色谱法分析其脂酸含量,检测了15名我国健康成人红细胞膜脂类的脂酸摩尔百分组成。结果表明:各脂类中,脂酸的类别基本相同,但其含量组成相差甚远。如磷脂酰乙醇胺(PE)富含C_(20:4);磷脂酰胆碱(PC)富含C_(18:2);神经鞘磷脂(SM)主要含C_(16:0);磷脂酰丝氨酸(PS)主要含C_(18:0);而以红细胞糖苷脂(GL)中脂酸含量最少。膜总脂中饱和脂酸与不饱和脂酸的含量大致相等,胆碱磷脂(PC+SM)的脂酸饱和度则明显高于氨基磷脂(PE+PS)。     

菜豆叶片衰老期间叶绿体被膜膜脂与脂肪酸组分的变化

菜豆叶片叶绿体总脂和被膜膜脂中均含有单半乳糖甘油二脂和双半乳糖甘油二脂,在整个衰老期间两种糖脂的比值变化不大。叶绿体总脂中含有5种磷脂,脂肪酸以不饱和的亚麻酸为主,而被膜膜脂中仅含磷脂酰胆碱和磷脂酰甘油,脂肪酸以饱和的棕榈酸为主,不饱和亚油酸为次。叶片衰老过程中被膜所含两种磷脂比值明显降低,脂肪酸的不饱和指数也因亚麻酸相对含量显著减少、棕榈酸相对含量增加而降低。     

Influence of plasmalogen deficiency on membrane fluidity of human skin fibroblasts: a fluorescence anisotropy study

The influence of plasmalogen deficiency on membrane lipid mobility was determined by measuring fluorescence anisotropy of trimethylammoniumdiphenylhexatriene (TMA-DPH) and diphenylhexatrienylpropanoylhydrazylstachyose (glyco-DPH) inserted in the plasma membranes of human skin fibroblasts deficient in plasmalogens. The cells used were from patients affected with cerebrohepatorenal (Zellweger) syndrome (CHRS) or rhizomelic chondrodysplasia punctata. Their plasmalogen content (0-5% of total phospholipid) is significantly reduced compared with that of control cells from healthy donors (13-15% of total phospholipid) or of CHRS fibroblasts supplemented with the plasmalogen precursor, hexadecylglycerol. Plasmalogen-deficient cells consistently showed lower fluorescence anisotropies of membrane-bound DPH fluorophores corresponding to higher membrane lipid mobilities as compared to controls. However, very similar lipid mobilities were found for sonicated aqueous dispersions of phospholipids extracted either from CHRS or control cells. Therefore, the differences observed with living cells are not due to differences in the overall physical properties of the membrane lipid constituents. Other phenomena such as lipid asymmetry and/or plasmalogen-protein interactions may be responsible for the effects observed in the biomembranes.     

Phospholipid aliphatic chain composition modulates lipid class composition, but not lipid asymmetry in Clostridium butyricum

The phospholipid composition of the butyric acid-producing clostridia is responsive to the degree of enrichment of the lipids with cis-unsaturated fatty acids. When Clostridium butyricum and Clostridium beijerinckii are grown on oleic acid in media devoid of biotin, the acyl and alk-1-enyl chains of the phospholipids become highly enriched with 18:1 and C19-cyclopropane. Under these conditions there is a marked increase in the glycerol acetals of the major plasmalogens of these organisms. We have grown both species on mixtures of palmitate and oleate in the absence of biotin. The alk-1-enyl chains were highly enriched with C18-unsaturated and C19-cyclopropane residues at all but the highest ratios of palmitate to oleate (80:20, w/w) added to the medium. At ratios of palmitate to oleate greater than or equal to 40:60, the saturated acid was incorporated predominantly into the phospholipid acyl chains in both organisms. The effects of increasing unsaturation of the acyl chains as the ratio of oleate to palmitate was increased was examined in C. butyricum. In cells grown on mixtures of palmitate and oleate equal to or exceeding 40% palmitate, the ratio of glycerol acetal lipid to total phosphatidylethanolamine (PE) was relatively constant. As the proportion of oleic acid added to the medium was increased, the ratio of glycerol acetal lipid to PE increased from 0.7 to 2.0. Thus the ratio of the polar lipids appears to respond to the content of phospholipids that contain two unsaturated chains. The fraction of PE present as plasmalogen remained relatively stable (0.82 +/- 0.05) at varying ratios of medium oleic and palmitic acids. Both the glycerol acetal of ethanolamine plasmalogen, and ethanolamine plasmalogen, are shown to be 80% or more in the outer monolayer of the cell membrane. These two polar lipids represent approx. 50% of the phospholipids in cells grown on exogenous fatty acid. The bulk of the remainder is polyglycerol phosphatides. We suggest that the ability of both species to grow with highly unsaturated membranes is related to their ability to modulate their polar lipid composition.     

营养盐初始组合浓度对类波氏真眼点藻生长和油脂积累的影响

为了研究培养基中主要营养元素氮、磷、硫初始组合浓度对类波氏真眼点藻(Eustigmatos cf.polyphem D.J.Hibberd)生长、油脂积累和脂肪酸组成的影响,分别以NaNO_3、K_2HPO_4、MgSO_4为氮、磷、硫源,设置不同的营养盐初始组合浓度对其进行培养,采用干重法、重量法、气相色谱分析法、元素分析仪等依次对生物量、总脂、脂肪酸组成和细胞内元素含量进行测定。结果显示,氮、磷、硫三者的初始组合浓度变化对类波氏真眼点藻的生长和油脂积累具有明显的影响,9 mmol/L NaNO_3、0.92 mmol/L K_2HPO_4、0.3 mmol/L MgSO_4营养条件下其生物量达到最大值,为9.19 g/L;低氮、低磷胁迫或二者共同胁迫均促进其油脂积累,但低硫胁迫对油脂的积累影响不大,最大油脂含量出现在3 mmol/L NaNO_3、0.029 mmol/L K_2HPO_4、0.3 mmol/L MgSO_4条件下,为68.7%(DW)。此外,类波氏真眼点藻富含棕榈油酸,占总脂肪酸含量的最大值达60.88%。选择合理的氮、磷、硫初始组合浓度能够有效促进其生长或油脂及棕榈油酸的积累。     

Lipid analysis of cells and chromatophores of Rhodopseudomonas sphaeroides

The phospholipids and fatty acid analysis of four strains of Rhodopseudomonas sphaeroides and of chromatophores from two strains show some differences and also show the presence of an unusual polar neutral lipid which is ninhydrin positive and which on acid hydrolysis yields ornithine and an unidentified amino compound. This lipid is called aminolipid-X and has a fatty acid composition very different from the phospholipids. Phosphatidylcholine (PC) and phosphatidylethanolamine (PE) contain a very small amount of plasmalogen forms as determined by combined mild alkaline hydrolysis, acetic acid hydrolysis and phospholipase A₂ hydrolysis.The reaction of intact cells and chromatophores with trinitrobenzenesulfonate (TNBS), fluorodinitrobenzene (FDNB) and isethionylacetimidate (IA) show that 78% of the total PE in chromatophores is localized on the outer membrane surface. In intact cells about 15–35% of the total PE is localized on the outer surface of the plasma membrane.     

Mature and immature synaptosomal membranes have a different lipid composition

Subfractionation of the optic tectum in chick embryos results in the isolation of two fractions enriched in synaptosomes (fraction A and fraction B). In chicks after hatching, this fractionation results in the isolation of a single synaptosomal fraction (fraction B) and of a fraction enriched in myelin membranes devoid of synaptosomes (fraction A). The lipid composition of synaptosomal fractions (A and B) and corresponding synaptosomal plasma membranes has been analyzed and compared to the lipid composition of similar fractions isolated from 2–3 day-old chicks. The phospholipid composition of fraction A in embryos was mainly represented by phosphatidylcholine (PC) and phosphatidylethanolamine (PE). The PE content was significantly lower than that of PC, which accounted for by approximately 50%. Sphingomyelin (SP) and phosphatidylinositol (PI) accounted for by only 6% of the total membrane phopsholipids. Fraction A isolated from the young chicks showed many significant changes. PC accounted for by approximately 40% and PE made up 35%. The amount of phosphatidylserine (PS) and SP increased. These data parallel our previous morphological observations, which showed that fraction A contains immature synaptosomes in embryos but myelin membranes and no synaptosomes in the young chicks. Fraction B has been shown to contain synaptosomes at all stages considered. It possessed in embryos a lipid composition similar to fraction A, except that PC content was higher in young embryos. The analyses on membrane fractions confirmed these results. On the contrary, this fraction showed many significant changes after hatching. The content of PC was significantly reduced, PE/PC ratio was significantly increased as well as ethanolamine plasmalogen (PLE) content. The percentage of PS, PI and SP were increased. The composition of fatty acids of the total fraction of phospholipids was also examined. The results parallel the observations on phospholipid classes.     

氯化钠处理对原壳小球藻生长及油脂积累的影响

采用Basal培养基,通过光学显微镜、电子显微镜、激光共聚焦显微镜以及尼罗红染色定量等方法研究了不同浓度氯化钠(0、150、300、600 mmol/L)对小球藻属原壳小球藻的生长状态、脂滴分布、总脂含量的影响。结果表明,添加不同浓度的氯化钠对原壳小球藻的生长有明显的影响,随着氯化钠浓度的增加,小球藻的生长速度受到明显的抑制,600 mmol/L氯化钠处理时生长几乎完全被抑制。在显微镜下观察,可见氯化钠浓度的增加会导致小球藻聚集成团,这种现象在150 mmol/L和300 mmol/L氯化钠培养下比较明显;通过电子显微镜下观察,可以发现培养初期,随着氯化钠浓度的增加,小球藻细胞壁增厚,脂滴增多。通过尼罗红染色对脂含量进行定量,处理初期脂滴的合成量在600 mmol/L时最高,但到后期,随着藻生物量的增加,150 mmol/L和300 mmol/L处理下脂合成量逐渐升高,而对照小球藻脂合成量基本不变。稳定期后,从生物量(干重)和脂总量来看,300 mmol/L氯化钠培养处理的小球藻虽然生物量只有对照的73.55%,但是总脂含量却是对照的2.22倍,可见一定浓度的氯化钠处理一定时间可显著提高原壳小球藻的油脂含量。     

利用平板反应器大量培养高产油绿藻——尖状栅藻的生长和油脂积累规律

以新近分离的淡水绿藻--尖状栅藻(Scenedesmus acuminatus)为研究对象,将改良的BG-11培养基中的初始NaNO₃浓度降低为6.0mmol/L和3.6mmol/L,利用新设计的内置拉筋平板式光生物反应器对尖状栅藻(S. acuminatus)进行大量培养。测定不同时相的生物量、总脂含量、脂组分含量及脂肪酸组成和含量,分析尖状栅藻(S. acuminatus)大量培养时的生长和油脂积累规律。当初始NaNO₃浓度为6mmol/L时其最高生物量(6.27g/L)明显高于初始NaNO₃浓度为3.6mmol/L时的生物量(5.30g/L);而最高的总脂含量在初始NaNO₃浓度为3.6mmol/L时获得为干重的56.6%,高于初始NaNO₃浓度为6mmol/L时的总脂含量(51.6%)。总脂经硅胶柱层析分级后得到三种类型的脂组分:中性脂、糖脂和磷脂,随着培养时间的延长中性脂含量逐渐增加,培养至18d后,中性脂的含量分别达到总脂的 90.9%(6 mmol/L NaNO₃)和 92.0%(3.6 mmol/L NaNO₃)及干重的 47.5%(6.0 mmol/L NaNO₃)和 51.4%(3.6 mmol/L NaNO₃)。主要脂肪酸组成为棕榈酸、棕榈油酸、硬脂酸、油酸、亚麻油酸和亚麻酸,这六种脂肪酸在不同时相的含量变化范围分别为89.92%~96.18%(占总脂肪酸)和12.5%~50.7%(占细胞干重)。总脂、中性脂及总脂肪酸单位体积产率分别为:0.18 g/L/d,0.16 g/L/d和0.15 g/L/d(6.0 mmol/L NaNO₃)及0.16 g/L/d,0.15 g/L/d和0.15 g/L/d(3.6 mmol/L NaNO₃)。研究结果表明,尖状栅藻(S. acuminatus)是一株易于规模化培养、脂肪酸组成适合于生物柴油生产的高产油微藻。     

Serine and ethanolamine incorporation into different plasmalogen pools: Subcellular analyses of phosphoglyceride synthesis in cultured glioma cells

In cultured glioma cells, plasma membrane (PM) is enriched in phosphatidylserine (PtdSer) and plasmalogens (1-O-alk-1-enyl-2-acyl-sn-glycero-3-phosphoethanolamine). Serine can be a precursor of headgroups of both ptdSer and ethanolamine phosphoglycerides (PE) including plasmalogens and non-plasmalogen PE (NP-PE). Synthesis of phospholipids was investigated at the subcellular level using established fractionation procedures and incorporation of [³H(G)]L-serine and [1,2-¹⁴C]ethanolamine. Specific radioactivity of PtdSer from [³H]serine was 2-fold greater in PM than in microsomes, reaching maximum by 2–4 h. Labeled plasmalogen from [³H]serine appeared in PM by 4 h and increased to 48 h, whereas almost no plasmalogen accumulated in microsomes within 12 h. In contrast, labeled plasmalogen from [1,2-¹⁴C]ethanolamine appeared in both PM and microsomes at early incubation times and became enriched in PM beyond 12 h. Thus, in glioma cells: (1) greater and faster accumulation of labeled PtdSer in PM may reflect direct synthesis from serine within PM; (2) PM is a major source of PtdSer for decarboxylation and PE synthesis; (3) NP-PE in both PM and microsome provides headgroup for synthesis of plasmalogen; and, (4) plasmalogen synthesis may involve different intracellular pools depending on headgroup origin.Abbreviations NP-PE nonplasmenylethanolamine phosphoglycerides including both diacyl and alkylacyl species - PE total ethanolamine phosphoglycerides: plasmalogen-plasmenylethanolamine or alkenylacyl ethanolamine phosphoglyceride (1-O-alk-1-enyl-2-acyl-sn-glycero-3-phosphoethanolamine) - PL phospholipid - PM plasma membrane - PtdCho phosphatidylcholine - PtdSer phosphatidylserine     

光径及氮浓度对缺刻缘绿藻生长、油脂和花生四烯酸积累的影响

以缺刻缘绿藻(Parietochloris incisa)为实验材料, 采用BG-11培养基, 分别在2种氮浓度和3种不同光径(LP)的柱状和平板光生物反应器中进行培养, 并探究其生长、油脂和花生四烯酸(AA)的积累规律。结果显示: 在两种光生物反应器中, 光径越小, 越有利于缺刻缘绿藻的生长。其中, 最大生物量均在17.6 mmol/L氮浓度时获得, 分别为5.09 g/L(2.5 cm-柱状)和2.98 g/L(3.0 cm-平板); 而最高油脂和AA绝对含量则均在1.0 mmol/L氮浓度和最大光径处获得, 分别为39.23%、13.21%(6.0 cm-柱状)和40.74%、11.33%(5.0 cm-平板); 另外, 两种光生物反应器中的最大油脂单位体积产率分别可以达到216.39 mg/(L·d)(17.6 mmol/L; 2.5 cm-柱状)和135.93 mg/(L·d)(1.0 mmol/L; 1.5 cm-平板); 而最高的AA单位体积产率均在1.0 mmol/L低氮条件, 最大光径处达到最大, 分别为21.65 mg/(L·d)(6.0 cm-柱状)和19.42 mg/(L·d)(5.0 cm-平板)。因此, 根据实际生产需要, 在1.0 mmol/L低氮条件下, 选择6.0 cm光径的柱状光生物反应器或5.0 cm光径的平板光生物反应器, 培养缺刻缘绿藻生产AA, 能有效降低生产成本。     

有机碳源对三角褐指藻生长、胞内物质和脂肪酸组分的影响

研究了3种有机碳对三角褐指藻生长、胞内物质和脂肪酸组分的影响。结果表明, 三角褐指藻具有利用有机碳进行兼养生长的能力, 生长速率加快, 倍增时间缩短, 生物量显著提高, 100 mmol/L甘油兼养的生物量最高(713 mg/L), 是自养(460 mg/L)的1.60倍, 乙酸钠和葡萄糖兼养的生物量分别是自养的1.28倍和1.21倍。兼养下蛋白质含量较自养明显下降, 碳水化合物和总脂含量高于自养, 乙酸钠和甘油兼养的总脂含量分别是自养的1.43倍和1.20倍, 葡萄糖兼养的总脂含量与自养无明显差异。3种有机碳兼养的饱和脂肪酸和单不饱和脂肪酸占总脂肪酸的比例增大, 多不饱和脂肪酸比例降低, EPA(eicosapentaenoic acid)比例降低, 乙酸钠兼养的胞内EPA含量(6.23%)和产量(36.59 mg/L)均高于自养, 分别是自养的1.10倍和1.40倍, 甘油和葡萄糖兼养的EPA含量和产量均低于自养。