李斯特菌溶血素基因的原核表达及其生物学特性

李斯特菌溶血素(LLO)是产单核细胞李斯特菌的主要毒力因子,利用PCR技术从血清型4b的产单核细胞李斯特菌菌株中扩增出编码LLO的hly基因,经克隆筛选和测序鉴定后,构建成该基因的原核表达质粒pGEX6P1hly,SDSPAGE结果表明：LLO与谷胱甘肽在大肠杆菌中已融合表达,融合蛋白的分子量为82kD;溶血实验证明融合蛋白具有较强的裂解真核细胞膜的作用,表明表达产物LLO具有生物活性,其溶血效价达2.26×101.4 HU/mg,这为进一步研究其致病与免疫机理、单抗研制和疫苗设计提供了条件。     

SC9-4和棕榈油的比较分析研究

SC_9-4是以植物油为原料研制成功的一种高速冷轧工艺润滑油。经武钢高速冷轧机轧制性能鉴定,它的高速轧制性能优于棕榈油。我们使用高效液相色谱比较分析鉴定了SC_9-4和棕榈油的甘油三酯组成。分析结果:SC_9-4:的主要甘油三酯是POP(42.8％)和PPP(14.6％),而棕榈油的主要甘油三酯是POP(31.3％)、POO(20.1％)、PLP(12.2％)和PLO(11.6％)。我们初步探讨甘油三酯组成与轧制性能关系认为,SC_9-4的高速轧制性能优于棕榈油,可能是和SC_9-4含PPP、PPS和PSS的含量(18.8％)高于棕榈油(9.9％)有关。     

不同脂肪源对细鳞鲑生长、脂质代谢及抗氧化性能的影响

配制4种含有鱼油(FO)、菜籽油(CO)、棕榈油(PaO)和大豆油(SO)的饲料投喂初始体重为(22.19±1.10) g的细鳞鲑幼鱼52d, 探讨不同脂肪源对细鳞鲑(Brachymystax lenok)幼鱼生长、脂质代谢及抗氧化性能的影响。结果显示, 增重率、特定生长率和饲料效率均以鱼油和大豆油组最高, 棕榈油组次之, 菜籽油组最低, 其中大豆油组与鱼油组无显著差异(P>0.05)。不同脂肪源对细鳞鲑幼鱼肌肉粗成分无显著影响(P>0.05)。摄食大豆油的细鳞鲑可以明显提升肌肉中的22:6n-3含量, 表明细鳞鲑具有将18:3n-3转化为22:6n-3的能力, n-3 系列脂肪酸是细鳞鲑起主要作用的必需脂肪酸。与鱼油相比, 植物油脂导致了细鳞鲑血清中甘油三酯含量的显著升高以及胆固醇含量的显著降低, 肝脏中ACC1和FAS的mRNA表达量明显下调, 以及Δ6 Fad的mRNA表达量的明显上调(P<0.05)。投喂不同脂肪源饲料的细鳞鲑肝脏ROS、MDA、GPx含量以及TBARS值发生明显变化(P<0.05)。研究结果表明鱼油和大豆油可作为细鳞鲑幼鱼饲料的脂肪源, 而棕榈油和菜籽油不适宜作为细鳞鲑幼鱼的单一脂肪源。     

油瓜油的甘油三酯组成研究

油瓜的种仁油可供食用。应用高效液相色谱技术,从油中分离分析出12种甘油三酯,其主要甘油二醋是甘油二亚油酸—棕榈酸酯(PLL)、甘油二棕榈酸—亚油酸酯(PLP)、甘油—棕榈酸—亚油酸—油酸酯(PLO)和甘油二棕榈酸—油酸酯(POP)。     

单增李斯特菌溶血素O的PEST序列激活ERK1/2磷酸化的作用

[目的]本研究旨在构建单核细胞增多性李斯特菌(Listeria monocytogenes,简称单增李斯特菌)溶血素O(Listeriolysin O,LLO)的关键结构域PEST序列(包含S44、S48和T51关键磷酸化位点)突变体,并针对其生物学功能展开研究。[方法]以李斯特菌参考菌株EGD-e为模板扩增编码LLO的hly基因,克隆至pET30a(+)原核表达载体,在此基础上利用氨基酸突变技术获得表达PEST突变体(LLO△PEST、LLOS44A、LLOS48A和LLOT51A)的重组质粒,转入E.coli Rosetta感受态细胞中,诱导表达重组蛋白经镍离子亲和层析纯化后进行SDS-PAGE分析。利用红细胞裂解试验检测重组蛋白的溶血活性,并通过Western blotting检测重组突变蛋白刺激Caco-2细胞后对MAPK关键信号分子ERK1/2磷酸化水平变化的影响。[结果]结果显示,本研究成功获得重组LLO及其突变体蛋白LLO△PEST、LLOS44A、LLOS48A和LLOT51A。在pH5.5和7.4条件下,LLO△PEST、LLOS44A、LLOS48A和LLOT51A均具有和LLO相当的溶血活性,说明PEST序列缺失或突变并不影响LLO的膜裂解活性。研究进一步发现,重组LLO及其突变蛋白刺激Caco-2细胞后均能激活ERK1/2的磷酸化。[结论]研究表明LLO的关键结构域PEST序列对于维持该蛋白的膜裂解能力及穿孔活性并非必需,且该结构域的缺失不影响李斯特菌在感染宿主时依赖LLO介导ERK1/2磷酸化的生物学过程。本研究将为进一步探索细菌感染过程中PEST序列对于LLO发挥生物学功能的潜在作用及分子机制奠定基础。     

在5种食用油中野菊花挥发油的抗氧化能力研究

采用水蒸气蒸馏法提取野菊[Dendranthema indicum (L. ) Des Moul. ]干燥头状花序的挥发油,以常用抗氧化剂二丁基羟基甲苯(BHT)为对照,并以油脂的过氧化值(POV)为指标,对野菊花挥发油在猪油、花生油、葵花籽油、玉米油和大豆油中的抗氧化能力进行了比较分析.结果表明,在供试的5种食用油中,各处理组(50.0 g食用油中野菊花挥发油添加量分别为0.005、0.025、0.050、0.075和0.100 g)的POV值均随贮藏时间(猪油和大豆油为28 d,花生油、葵花籽油和玉米油为35 d)的延长逐渐增大;随挥发油添加量的增加,5种食用油的POV值总体上呈逐渐减小的趋势.在28 d的贮藏期内,仅添加了0.075～0.100 g挥发油的玉米油和大豆油的POV值极显著低于对照(0.010 g BHT)和空白;在贮藏的前中期,仅添加了0.075～0.100 g挥发油的花生油和葵花籽油的POV值极显著低于对照和空白,但在贮藏的中后期花生油和葵花籽油各处理组的POV值则极显著高于对照;添加了挥发油的猪油在28 d的贮藏期内其POV值基本上极显著高于对照但低于空白.说明在玉米油和大豆油中添加适量的野菊花挥发油具有较强抗氧化能力,且抗氧化效果总体上高于BHT;而添加适量的野菊花挥发油也能在一定程度上延缓花生油和葵花籽油的氧化过程,但抗氧化效果低于BHT;而添加适量的野菊花挥发油仅能部分缓解猪油的氧化过程,其抗氧化效果明显低于BHT.研究结果显示,在不同的食用油中野菊花挥发油均有一定的抗氧化作用,但其抗氧化能力有一定的差异,这种差异与食用油种类、野菊花挥发油的添加量及食用油的贮藏时间均有一定的关系.     

植物油中酸价的测定及结果分析

植物油是人们生活中不可缺少的物品,油质的好坏直接影响着饮食口感和身体健康。生活中常见的植物油有大豆油、花生油、菜籽油等,其中酸价指标是植物油各项指标中最为重要的一项卫生指标,通过植物油中酸价大小的测定,可以分析出植物油品质的好坏。     

饲料中不同脂肪源对黄鳝生长和组织中脂肪酸含量的影响

用6种不同脂肪源:鱼油(FO)、亚麻油(LO)、大豆油(SO)、二十碳四稀酸(ARA)+花生油(APO)、花生油(PO)和猪油(PL),配制了6组含脂量为8%的等氮(44%粗蛋白)、等能(19.4 MJ/kg)饲料,喂养黄鳝10周后,其结果显示:饲料中不同脂肪源对黄鳝的生长有影响。PO组的特定生长率(SGR)最低,显著低于SO、FO组(P<0.05);SO组的SGR显著高于除FO组之外的其他各组(P<0.05)。黄鳝组织中的18:3n-3和18:2n-6含量与饲料中的对应脂肪酸含量呈线性相关,缺乏ARA的试验组黄鳝各组织中ARA的含量与饲料中的18:2n-6含量存在弱的线性相关;但在各组织中ARA的含量均显著低于APO组(P<0.05)。缺乏二十碳五烯酸(EPA)、二十二碳六烯酸(DHA)的LO、SO、PL、PO各组,卵巢中的EPA和DHA含量与饲料中的18:3n-3含量呈对数函数相关;但在各组织中EPA、DHA的含量均显著低于FO组(P<0.05)。结果表明n-3多不饱和脂肪酸(PUFA)是黄鳝生长重要的脂肪酸,黄鳝生长需要一个适宜的n-6/n-3比值;大豆油、亚麻油、猪油替代鱼油可基本满足黄鳝生长对脂肪酸的需要,其中以大豆油最佳。黄鳝可合成长链多不饱和脂肪酸(LC-PUFA),但合成LC-PUFA的量有限。     

单核增生性李氏杆菌溶血素的原核表达及其单克隆抗体的制备

为了深入研究单核增生性李氏杆菌(LM)致病机理,从其基因组中克隆李氏杆菌溶血素基因hly,并将其与原核表达载体连接在大肠杆菌BL21中表达携带His标签的李氏杆菌溶血素(LLO)融合蛋白,经镍柱纯化得到重组LLO蛋白作为免疫原并免疫小鼠。取免疫小鼠的脾细胞与骨髓瘤细胞(Sp2/0)进行融合,经过3次亚克隆后获得3株稳定分泌针对LLO蛋白单抗的杂交瘤细胞株,分别命名为Anti-LLO1、Anti-LLO2、Anti-LLO3;经ELISA测定其细胞培养上清效价分别为1:3.6×104、1:6.4×104、1:1.6×104,腹水效价分别为1:2×107、1:2×107、1:1×107;亲和力解离常数(Kd)分别为6.18×10-11、7.50×10-11、6.27×10-11;3株单抗的IgG亚类均为IgG1。经Westernblotting鉴定证明,该3株抗体均能特异地识别李氏杆菌LLO蛋白,该单抗的制备为深入研究LM的致病机理奠定了基础。     

转基因与非转基因大豆油主要营养成分比较分析

为了解消费者对转基因食品的认知消费情况,在河南地区消费者调研的基础上,对转基因和非转基因大豆油的主要营养成分进行了比较分析。结果表明：63.41%的被调查者听说过转基因食品,其中认为转基因食品对人体健康和环境影响长期短期均有害的占36.22%、自己不会购买转基因食品的占64.4%。转基因和非转基因大豆油在脂肪酸、维生素E和甾醇的种类上一致,其中非转基因大豆油中α-亚麻酸的含量显著高于转基因大豆油,其余各组分含量在两者中差异不显著;两者脂肪酸总量中不饱和脂肪酸占比超过80%,β-谷甾醇在甾醇中的含量最高。综上,消费者对转基因食品认知度、关注度较高,转基因和非转基因大豆油在主要营养成分上差异不明显。本研究为转基因食品的安全性评价和政府制定监管政策提供科学依据。     

Continuous enzymatic transesterification of sesame oil and a fully hydrogenated fat: effects of reaction conditions on product characteristics

An immobilized lipase from Thermomyces lanuginosus (TL IM) was employed to mediate the continuous transesterification of sesame oil and fully hydrogenated soybean oil (FHSBO) in a packed-bed reactor operating at 70 degrees C. Reactions between sesame oil (rich in LLL (15.97%), LOL (31.56%), and OLO (21.15%) [L = linoleic; O = oleic]) and the fully hydrogenated fat ((73.7% SSS, 26.3% SPS) [S = stearic; P = palmitic]) produced semi-solid fats. These products are complex mixtures of triacylglycerol (TAG) species whose compositions depend on reaction conditions. The dependence of the steady state product TAG profile on space time was determined for four initial weight ratios of sesame oil to hydrogenated fat (90:10, 80:20, 70:30, and 60:40). Except for the trial involving a weight ratio of sesame oil to FHSBO of 60:40, near equilibrium conditions were achieved at space times of 30 min-1 h. The chemical, physical, and functional properties of the product semi-solid fats were characterized. The predominant TAG species in the quasi-equilibrium products obtained from the mixture initially containing 90% (w/w) sesame oil and 10% FHSBO were LOL (26.22%) and OLO (21.92%). For transesterification of 80% sesame oil and 20% FHSBO, the major product species were OOP (21.27%), LOL (17.46%), and OLO (13.93%). OOP (24.38%) was the major product for reaction of 70% sesame oil with 30% FHSBO. Appropriate choices of reaction conditions and initial ratios of sesame oil to FHSBO lead to TAG with melting profiles and solid fat contents (SFC) similar to those of a variety of commercial products.     

桃、李、杏种仁油的甘油三酯组成研究

李(Prunus salicina L.)、桃(Amygdalus persica L.)、杏(Armeniaca vulgarisLam.)的种仁富含油脂,主要脂肪酸成分为油酸和亚油酸。近年 Geeraert 和 Sandra报道了杏仁油的甘油三酯组成的气相色谱图。桃、李仁油的甘油三酯,以及甜杏仁油与苦杏仁油的比较,均未见报道。本文报道这几种水果的种仁成分。1.试验材料和油样提取苦杏仁和甜杏仁为商品;李仁和桃仁来源于广东地区栽培品种。除甜杏仁外,其余种仁均有苦味。种仁捣碎,石油醚(30—60℃)室温浸提,减压蒸除溶剂得分析油样。除甜杏仁油外,其余均有苦杏仁甙的气味。     

Triglyceride metabolism in germinating Andropogon gayanus seeds

Seed triglycerides of Andropogon gayanus contained 17 fatty acid moieties, principally palmitic, oleic and linoleic acids. These were distributed in an essentially random manner amongst the triglycerides to form the following main types: POL, PLL, OOL, LLO and LLL. Triglycerides decreased during both light and dark germination but there was no evidence for selective hydrolysis. Free fatty acids appear to be derived from triglyceride hydrolysis but the free and triglyceride fatty acid composition differed. Less palmitic, oleic and linoleic acids and more stearic, linolenic and C₂₀-acids were found in the free state than combined in the triglycerides. Free fatty acids did not accumulate during germination.     

Effects of Triterpenes on the Mutagenicities of Various Mutagens toward Salmonella

The effects of different dietary levels of gamma-linolenic acid (GLA) on lipid metabolism was studied in rats using a combination of evening primrose oil (EPO) and palm oil (PLO). EPO compared to PLO significantly reduced liver cholesterol and triglyceride after 4 weeks of feeding, and the effect remained even when EPO was mixed with PLO at the same weight basis. The serum triglyceride level also tended to be low on feeding EPO. Neither liver Δ6-desaturase and phospholipase A₂ activities nor aortic production of prostacyclin and thromboxane A₂ production by platelets were influenced significantly by the fat type, suggesting a peculiar effect of PLO. The percentage of arachidonic acid in liver, serum, and aortic phosphatidylcholine depended on the dietary level of GLA. A more distinct increase in arachidonic acid was observed in tissue triglycerides of rats fed EPO. GLA appears to exert favorable effects on lipid metabolism even when the P/S ratio was lowered from 13.7 of EPO to 1.8 of the 1:1 mixture of EPO and PLO.     

A Study on the Triglyceride Composition in Apricot Kernel Oil,Peach Kernel Oil and Plum Kernel Oil

The triglyceride composition of kernel oils from apricot, peach and plum was separated and analysed by reverse phase high performance liquid chromatography (RP-HPLC). The results showed that their oils predominantly contained OOO and OLO among 11 triglycerides.     

Triacylglycerols composition, oxidation and oxidation compounds in camellia oil using liquid chromatography-mass spectrometry

Camellia seed oil is one of most important edible oil, rich in oleic acid and contains many natural antioxidants with various biological activities. During preparation of foods or storage camellia oil oxidizes by the auto-oxidation and produce oxidized compounds. Traditional analytical techniques like FFA, POV are used for the determination of oxidation and adulteration of oils and fats. These methods were rarely able to detect the oxidized compounds produced and extent of oxidation. This paper presents the uses of liquid chromatography coupled to electrospray ionization mass spectrometry (HPLC-ESI-MS) for the analysis of triacylglycerols (TAGs) composition and evaluation of auto-oxidation and oxidation products of camellia seed oil. The camellia oil was auto-oxidized for 12 months at room temperature. The TAGs were identified from their characteristics fragmentations such as protonated molecular ion, ammonium and sodium adducts, diacylglycerols, epoxy-diacylglycerols fragments and mono-acylglycerol fragments in ESI-MS mass spectra. HPLC-ESI-MS data revealed the separation and identification of 15 TAGs. The major TAGs separated and identified in camellia seed oil were POO, OOO, OLO, PLO/POL, OLL, SOO, ALO and OLLn. The auto-oxidation studies revealed a total loss of LnLLn, LnOLn, LLLn and OLLn amounting about 13.5% total oxidation. The auto-oxidation products were epoxy hydroperoxides, epoxy epidioxides, and mono-epoxides. It was observed that these were characteristic compounds produced in high oleic oils.     

Differences in the Rates of Metabolism of Various Triacylglycerols during Seed Germination and the Subsequent Growth of Seedlings of Dioscorea tokoro, a Perennial Herb

Differences in the rates of metabolism of various triacylglycerols(TAGs) in the reserve oil of seeds during the course of germinationand seedling growth were studied in Dioscorea tokoro, an EastAsian perennial herb. Eleven molecular species of TAG were determinedin the reserve oil of seeds. These TAGs contained five fattyacids: palmitic acid (P); stearic acid (S); oleic acid (O);linoleic acid (L); and linolenic acid (Ln). At 20°C, levelsof TAGs with one or less than one L moiety, such as OLO, OOO,PLO, POO, OLS, and OLnO, decreased rapidly during germinationand the initial elongation of seedlings, whereas levels of otherTAGs decreased slowly during the subsequent development of seedlings.Even though seeds were unable to germinate at these temperatures,levels of some TAGs decreased during incubation at 5°C andat 30°C. Such changes might be related to the metabolismrequired for the survival of seeds at these temperatures. Itis also possible that they might be involved in the breakingof dormancy at low temperatures or in the induction of secondarydormancy at high temperatures. The present results provide anexample of the selective utilization of the TAGs in the reserveoil of seeds during germination and the subsequent growth ofseedlings. (Received October 3, 1995; Accepted January 31, 1996)     

A Study on the Main Fatty Acids and Triglyceride Composition of Kernel Oil of Iodes vitiginea

The kernel of lodes vitiginea (Hance) Hemsl. (Icacinaceae) contains 39---50% of oil. Analysing with GC, the main fatty acids are hexadecenoic and palmitic acid. By means of ozonization, GC/MS, IR, etc., the hexadecenoic acid was identified as cis- 7-hexadecenoic acid. Thirteen triglycerides (peak) have been separated by HPLC from oil, among them, eight triglycerides were identified by C,C with HHH, 18.4; HHP, 32.3%; PHP, 16.8% as the main composition.