菠萝叶膜脂脂肪酸组成的温度效应及其与抗寒性的关系

生长在不同季节的菠萝叶膜脂脂肪酸的配比存在着明显差异;随着大气温度的下降,18:1含量显著减少,18:2和18:3含量增加。不同品种均表现出一致的变化趋势。致害低温破坏了膜脂,使较不抗寒品种的16:0含量增加,18:2和18:3含量减少;较抗寒品种这种变化则较不显著。适当低温锻炼能改变膜脂脂肪酸的代谢过程,16:0和18:1含量减少,18:3含量增加。当处于更低温度时,除了16:0和18:1继续减少外,有一部分18:2也脱饱和而转变为18:3。因之明显地增加了膜脂中18:3的含量和脂肪酸的不饱和度,从而有利于抗寒性的提高。而品种间的抗寒性差异亦是在此低温期间表现出来。     

生物科学中一个崭露头角的领域：高静压力研究

通过介绍高静压力作用于生物大分子的机制,提出一些有关高压力生物学的基本概念。此外,还介绍了高静压力在研究蛋白质构象,蛋白质－蛋白质、蛋白质－核酸、蛋白质－配基等相互作用,酶活性的调制以及在生物技术中的应用,表明高压力是一很好的研究手段。     

基于膜压力应答途径的大肠杆菌丁醇耐受性

【目的】筛选丁醇压力下Escherichia coli中参与溶剂压力应答的细胞信号传导途径,并从应答途径出发,提高E.coli丁醇耐受性。【方法】在丁醇压力下,利用RT-PCR分析大肠杆菌内膜压力应答途径中反应调节因子(response regulator,RR)的表达水平,通过Red同源重组以及一步克隆的方法分别构建外膜脂蛋白Nlp E和分子伴侣蛋白Spy的敲除菌株E.coli JM109(Δnlp E)和E.coli JM109(Δspy)及重组菌株E.coli JM109/p QE80L-nlp E和E.coli JM109/p QE80L-spy,并测定其溶剂耐受性和细胞膜疏水性。【结果】0.8%(V/V)丁醇处理10 h后,Cpx和Bae双组分压力应答途径中的cpx R和bae R基因的表达水平分别提高了8.3和3.3倍;分别在含0.6%(V/V)四氢呋喃、0.1%(V/V)甲苯和0.6%(V/V)环己烷的培养基中培养10 h后,重组菌株E.coli JM109/p QE80L-spy和E.coli JM109/p QE80L-nlp E的OD600相比对照组(OD600增长0.02-0.04)分别增长了0.13-0.17和0.05-0.13,重组菌的溶剂耐受性得到了显著提高。【结论】Cpx和Bae系统参与大肠杆菌丁醇压力应答,分子伴侣蛋白Spy的过表达能够有效提高大肠杆菌对有机溶剂的耐受性,本研究为阐明微生物有机溶剂耐受性机制提供了理论依据。     

温度、光照和盐度对2株曼氏骨条藻生长及脂肪酸组成的影响

研究温度、光照、盐度对2株曼氏骨条藻（Skeletonema munzelii）SM-1、SM-2生长、总脂含量及脂肪酸组成的影响,以确定其生长及油脂、多不饱和脂肪酸积累的最适生态条件。在实验室智能光照培养箱内不充气培养控制条件下,采用单因子试验分别研究了不同温度（10、15、20、25和30℃）、光照强度（20、40、60、80、100和120μmol/m2·s）、盐度（10、15、20、25、30、35和40）对2株藻的生长、总脂含量及脂肪酸组成的影响。结果表明：不同温度、光照强度及盐度对2株藻的生长、总脂及脂肪酸含量影响均有显著影响（P〈0.05）。藻株SM-1生长的最适温度为25℃,最适光强60μmol/m2·s,最适盐度30,而低温（10~15℃）,低光照（20μmol/m2·s）,低盐度（盐度15）更有利于总脂及PUFA的积累。SM-2生长的最适温度为20℃,最适光强60μmol/m2·s,最适盐度30,而低温（10~15℃）,低光照（20μmol/m2·s）更有利于其总脂及PUFA的积累,低盐（盐度15）则更有利于PUFA的积累。因此在实际生产中,2株藻可先在最适条件下培养以增加生物量,后转至利于PUFA积累的条件下提高PUFA产量。     

发菜膜脂及其脂肪酸组成

对野生发菜(Nostocflagelliforme Bom.et Flab)的膜脂(主要成分为类囊体膜脂)及其脂肪酸组成进行了测定分析.发菜的膜脂由单半乳糖甘油二酯(MGDG)、双半乳糖甘油二酯(DGDG)、磷酯酰甘油(PG)和硫代异鼠李糖甘油二酯(SQDG)组成,其酯酰基连接有棕榈酸(16:0)、十六碳烯酸(16:1)、硬脂酸(18:0)、油酸(18:1)、亚油酸(18:2)和亚麻酸(18:3)6种脂肪酸.发菜的不饱和脂肪酸含量可达总脂的73%,特别是16:1和18:3分别高达29%和34%,远远高于已报道的其他蓝藻,说明了发菜类囊体膜具有较强的抗逆性特点.同时还对复水30 min和复水后生长24 h的发菜膜脂及其脂肪酸组成进行了分析.结果表明,复水对野生发菜的膜脂及其脂肪酸组成没有显著影响,说明发菜的膜脂和脂肪酸组成在干燥-吸水过程中能保持很高的稳定性.     

优化硫酯酶表达提高大肠杆菌脂肪酸乙酯的生物合成效率

利用基因工程大肠杆菌直接从头生物合成脂肪酸乙酯 (生物柴油) 的相关研究引起了国内外研究人员的广泛关注。在本课题组已经构建的能够从头合成脂肪酸乙酯的大肠杆菌菌株KC3的基础上,通过替换表达不同来源的硫酯酶,发现表达来源于香樟树的硫酯酶Cc FatB1基因能够提高脂肪酸乙酯产量。进一步通过共表达Cc FatB1和大肠杆菌硫酯酶tesA’基因,以及启动子优化,获得了高产脂肪酸乙酯工程菌株KC4。KC4菌株在摇瓶条件和发酵条件下的单位生物量脂肪酸乙酯产率分别为21.4 mg/ (L?OD600)和31.16 mg/ (L?OD600)。该工程菌株的构建进一步提高了脂肪酸乙酯产量,显示了通过基因工程改造大肠杆菌从头合成生物柴油的应用潜力。     

不同脂肪源对异育银鲫的生长、体组成和肌肉脂肪酸的影响

配制了十种等氮等能的饲料饲喂3.53 g的异育银鲫幼鱼12周, 探讨异育银鲫对不同脂肪源的利用效果。十种饲料中分别添加8%的鱼油(FO)、椰子油(CNO)、玉米油(CO)、亚麻油(LO)、大豆油(SO)、菜籽油(RO)、1∶1鱼油-椰子油(FCNO)、1∶1鱼油-玉米油(FCO)、1∶1鱼油-亚麻油(FLO)和1∶1∶1∶1鱼油-椰子油-玉米油-亚麻油混合油(MIX)。每组饲料三个平行, 每个平行30尾。实验在循环水养殖系统中进行, 水温控制在(241)℃。结果表明, 在单一脂肪源中, 豆油组和椰子油组的增重率最高, 其次是菜籽油组, 鱼油、玉米油和亚麻油组的增重率最低。与相应的单一脂肪源相比, 饲料中鱼油与椰子油、玉米油或亚麻油1∶1混合后使用提高了异育银鲫的生长。摄食不同脂肪源饲料的异育银鲫血清生化指标、各组织的水分和脂肪含量差异不明显(P0.05)。肌肉脂肪酸与饲料脂肪源呈明显正相关。摄食豆油和菜籽油饲料的鱼体肌肉中20:4n-6较高, 而摄食亚麻油饲料的鱼则含有较高的20:5n-3和22:6n-3, 表明异育银鲫具有转化18:2n-6和 18:3n-3为高不饱和脂肪酸的能力。从实验可以看出, 豆油、椰子油和菜籽油是异育银鲫饲料中良好的脂肪源。       

特定电磁波对小麦幼苗膜脂肪酸组成的影响

本文研究了特定电磁波处理小麦种子对其幼苗叶片的总膜脂肪酸组成及线粒体膜脂肪酸组成的影响。在正常情况下,ＴＤＰ处理的总膜脂肪酸组成及线粒体膜脂肪酸组成与对照的区不大。     

Differential inactivation of glucose- and glutamate dependent acid resistance of Escherichia coli TMW 2.497 by high-pressure treatments

The inactivation by 200–400 MPa and post-pressure survival at acid conditions of E. coli TMW 2.497 was characterized by the measurement of intracellular pH (pH_in), viable cell counts, glutamate (Glu) and arginine (Arg) consumption, and the influence of mild adaptation to mild acid stress prior to pressure treatment. Glutamate and arginine did not affect viable cell counts or the pH_in during pressure application but improved the ability to maintain a high pH_in after pressure treatment. In pH 4.0 buffer without arg and glu, a 3 log reduction of cell counts occurred after 24 h of incubation, whereas little or no loss of viability was observed after 24 h incubation in the presence of glu and arg. During post-pressure incubation at pH 4.0, 10 mM glutamate were metabolized but only 2 mM arginine were used, indicating that glutamate rather than arginine was responsible for the protective effect on pH_in and survival. In conclusion, the pressure induced, irreversible loss of the transmembrane ΔpH correlates to cell death and glu stabilizes the pH_in of E. coli during post-pressure incubation.     

Involvement of phospholipids in resistance and adaptation of Escherichia coli to acid conditions and to long-term survival

In Escherichia coli membranes, three major phospholipids are formed: phosphatidylethanolamine (PE), phosphatidylglycerol (PG) and cardiolipin (CL). We report here the survival of mutants lacking either PE or both PG and CL at an acid pHo and during long-term survival experiments. Stationary phase cultures of E. coli lacking PE are much more sensitive to acid shock (pHo 3) than the wild-type strain. Moreover, in the strain lacking PE, long-term survival in stationary phase is impaired and after 5 days no viable cells are recovered. The survival of an exponential phase culture to acid shock is known to be increased if the culture is exposed to moderately acid conditions (pHo 5) prior to a shift to pHo 3. If either PE or both PG and CL are missing, the exposure to pHo 5 does not increase the survival at pHo 3.     

On-line fluorescence determination of pressure mediated outer membrane damage in Escherichia coli

The outer membrane (OM) of Gram-negative bacteria provides a protective barrier for natural occurring inhibitors. Pressure mediated OM permeabilisation therefore contributes to the elimination of Escherichia coli and Salmonella by pressure preservation processes. Pressure mediated inactivation, sublethal injury, and membrane permeabilisation of E. coli were determined using two strains differing in their barotolerance. Pressure treatment of E. coli TMW 2.427 at 300, 500 and 600 MPa for 40 min resulted in a 0, 1, and greater 6 log decrease of viable cell counts, respectively. The kinetics of OM and cytoplasmic membrane permeabilisation after pressure treatment were determined by staining of pressure treated cells with the fluorescent dyes propidium iodide (PI) and 1-N-phenylnaphtylamine (NPN), respectively. A slight increase of PI fluorescence was observed at conditions resulting in a greater 6 log decrease of viable cell counts only. In contrast, increased NPN fluorescence indicating OM permeabilisation was observed prior to cell death and sublethal injury. An on-line assay for determination of pressure mediated OM damage based on NPN fluorescence was established to distinguish between reversible and irreversible OM damage. Generally, the same degree of outer membrane damage was observed by either on line or off line determinations. However, whereas reversible membrane damage occurred fast and in thermodynamic equilibrium with pressure conditions, irreversible outer membrane damage was a time dependent process.     

大肠杆菌酸性磷酸酶基因克隆表达及应用

将大肠杆菌K-12的酸性磷酸酶(AphA)完整基因和去信号肽基因分别克隆到pET-28a(+)栽体上,并转化入大肠杆菌BL21( DE3)中.经诱导检测,重组菌均能表达出高活性的可溶性酶蛋白,去信号肽表达更稳定.对重组菌的活性研究表明,相对于野生菌,重组菌酶活力得到大幅度提高,同时,以pNPP、肌苷为底物进行磷酸转移催化反应,在pH4.0-6.0、反应温度37℃条件下,约有30％的肌苷可转化为IMP,但随着反应的进行所形成的IMP又被该酶降解,向反应液中加入EDTA即可明显抑制酶的水解活性,减缓IMP的降解速率.     

Effect of growth temperature on the fatty acid composition of Mycobacterium phlei

In Mycobacterium phlei, fatty acid unsaturation increased with decreasing temperature. The 10-hexadecenoic acid content increased as the temperature was reduced from 35°C to 26–20°C. At lower temperatures tuberculostearic acid decreased while oleic and linoleic acids increased, the latter being found in M. phlei for the first time. Concomitantly palmitic acid content decreased, and the 6- and 9-hexadecenoic acids increased slightly on reducing the temperature from 35 to 10°C. Thus, down to 26–20°C palmitic acid was mainly replaced by 10-hexadecenoic acid. From this range down to 10°C, palmitic and tuberculostearic acids were replaced by oleic and linoleic acids. Consequently, fatty acid branching decreased and mean chain length increased, as the temperature was reduced. These observations support the view that regulation of membrane fatty acid composition is part of microbial temperature adaptation, and that themechanism behind the responses might be more complex than generally believed.Abbreviations ACP acyl carrier protein - FAS I (Type I) fatty acid synthetase I - FAS II (Type II) fatty acid synthetase II - MGLP methylglucose containing lipopolysaccharide - MMP methylmannose containning polysaccharide     

The glutamate-dependent acid resistance system of Escherichia coli and Shigella flexneri is inhibited in vitro by L-trans-pyrrolidine-2,4-dicarboxylic acid

Strains of Escherichia coli K-12, O157:H7, and Shigella flexneri grown to stationary phase in complex unbuffered media can survive for several hours at pH 2.5. This stationary-phase acid resistance phenotype is dependent upon the alternate sigma factor sigmas and the supplementation of either glutamate or glutamine in the acidified media used for acid challenge. Acid resistance under these defined conditions can be inhibited by the glutamate analog L-trans-pyrrolidine-2,4-dicarboxylic acid which blocks uptake of glutamate/glutamine by selective inhibition. The gadC gene, encoding an inner membrane antiporter essential for the expression of acid resistance, could not be detected in other family members of the Enterobacteriacae.     

Effect of temperature on the ATP pool and adenylate energy charge in Escherichia coli

The effect of cultivation temperature on the ATP pool and adenylate energy charge (EC) in Escherichia coli has been studied in both batch and continuous cultures. In batch culture, μ_max and the ATP pool increased with increasing growth temperatures between 27–42°C (from 0.26 to 0.62 h⁻¹, and from 5.1 to 8.2 nmol/mg dry wt., respectively). In continuous culture at a constant dilution rate (D = 0.2 h⁻¹), with increasing growth temperatures between 28–43°C, the ATP pool increased about 2-fold (from 4.2 to 8.1 nmol/mg dry wt) and the EC from 0.80 to 0.99.     

Cell density dependent acid sensitivity in stationary phase cultures of enterohemorrhagic Escherichia coli O157:H7

Escherichia coli O157:H7, the causative agent of hemorrhagic colitis and hemolytic uremic syndrome, can survive in a highly acidic environment. The acid resistance of this organism, as measured by its ability to survive in low pH, depended on the density of the cells present during the assay. At low cell densities (相似文献