一株生物表面活性剂产生菌的分离及其特性研究

模炼油厂污泥中分离得到1株生物表面活性剂产生菌C-3, 根据其生理生化特性和16S rDNA序列相似性分析, 将其鉴定为铜绿假单胞菌(Pseudomonas aeruginosa)。初步研究了其产生物表面活性剂的最适条件, 在以植物油为碳源、30°C、初始pH 8、Ca2+浓度20 mg/L、250 mL三角瓶中装75 mL发酵液的条件下, 最利于菌株的生长和生物表面活性剂产生。它的成分为糖脂类物质, 临界胶束浓度(CMC)为50 mg/L, 具有很好的增溶效果。     

Targeting NADPH Oxidase and Phospholipases A2 in Alzheimer’s Disease

Alzheimer’s disease (AD) is marked by an increase in the production of extracellular beta amyloid plaques and intracellular neurofibrillary tangles associated with a decline in brain function. Increases in oxidative stress are regarded as an early sign of AD pathophysiology, although the source of reactive oxygen species (ROS) and the mechanism(s) whereby beta amyloid peptides (Aβ) impact oxidative stress have not been adequately investigated. Recent studies provide strong evidence for the involvement of NADPH oxidase and its downstream oxidative signaling pathways in the toxic effects elicited by Aβ. ROS produced by NADPH oxidase activate multiple signaling pathways leading to neuronal excitotoxicity and glial cell-mediated inflammation. This review describes recent studies demonstrating the neurotoxic effects of Aβ in conjunction with ROS produced by NADPH oxidase and the downstream pathways leading to activation of cytosolic phospholipase A₂ (PLA₂) and secretory PLA₂. In addition, this review also describes recent studies using botanical antioxidants to protect against oxidative damage associated with AD. Investigating the metabolic and signaling pathways involving Aβ NADPH oxidase and PLA₂ can help understand the mechanisms underlying the neurodegenerative effects of oxidative stress in AD. This information should provide new therapeutic approaches for prevention of this debilitating disease.     

长吻对鱼粉等六种商品饲料中粗蛋白和能量的消化率

本试验以三氧化二铬（Ｃｒ２Ｏ３）为指示物测定Ⅰ龄长吻对鱼粉、猪血粉、豆饼粉、菜籽饼粉、小麦粉和玉米蛋白粉等六种商品饲料原料粗蛋白和总能量的消化率。试验饲料由参考饲料和试验原料以７０∶３０的比例混合挤压制成颗粒饲料。试验鱼粪便样品用虹吸方式收集,并将测试的消化率结果经过降低１０％的校正。由此所得消化率值更能反应长吻的消化生理状况。另外本文对六种饲料的消化率结果进行了讨论。     

云南红豆杉紫杉烷2α-O-苯甲酰转运酶基因的克隆及定性

通过RACE技术,克隆了云南红豆杉紫杉烷2α-O-苯甲酰转运酶基因(TyTBT),该酶催化2-去苯甲酰-7,13-二乙酰巴卡亭Ⅲ生成7,13-二乙酰巴卡亭Ⅲ,是紫杉醇合成途径中的关键酶之一.TyTBT基因cDNA全长1481 bp,含有1 320 bp的开放读码框,编码440个氨基酸的多肽,分子量为50 050 Da,等电点为6.17.氨基酸序列比对表明TyTBT同植物酰化酶家族的其它成员有较高的相似性,超过67%,同东北红豆杉和曼地亚红豆杉的紫杉烷2α-O-苯甲酰转运酶氨基酸序列的一致性和相似性达到最高,分别为95%和96%.广泛地比对分析证明这种较高的相似性在红豆杉属的其它酶家族中具有普遍性,进化树分析表明同东北红豆杉和曼地亚红豆杉的紫杉烷2α-O-苯甲酰转运酶(TBT)的相似性高于紫杉醇合成途径中的其它酰化酶.     

A novel RGD-toxin protein,Lj-RGD3, from the buccal gland secretion of Lampetra japonica impacts diverse biological activities

RGD (Arg-Gly-Asp) motif toxin proteins from snake venoms, saliva glands secretion of leech or tick have typical characteristics of inhibiting platelet aggregation, angiogenesis, and tumor growth. Here we report cloning and characterization of a novel RGD-toxin protein from the buccal gland of Lampetra japonica. In an attempt to study the activities of anticoagulant in the buccal gland secretion of L. japonica, we established buccal gland cDNA library and identified a gene encoding a predicted protein of 118 amino acids with 3 RGD motifs. The predicted protein was named Lj-RGD3. We generated the cDNA of Lj-RGD3 and obtained the recombinant protein rLj-RGD3. The polyclonal antibodies against rLj-RGD3 recognized the native Lj-RGD3 protein in buccal gland secretion in Western blot analyses. The biological function studies reveal that rLj-RGD3 inhibited human platelet aggregation in a dose-dependent manner with IC₅₀ value at 5.277 μM. In addition, rLj-RGD3 repressed bFGF-induced angiogenesis in the chick chorioallantoic membrane model. rLj-RGD3 also inhibited the adhesion of ECV304 cells to vitronectin. Furthermore, rLj-RGD3 induced apoptosis and significantly inhibited proliferation, migration, and invasion evoked by bFGF in ECV304 cells. Taken together, these results suggested that rLj-RGD3 is a novel RGD-toxin protein possessing typical functions of the RGD-toxin protein.     

半导体矿物介导非光合微生物利用光电子新途径

自然界中微生物按其能量代谢途径主要分为两种:光能营养微生物和化能营养微生物.化能营养微生物作为非光能营养微生物长期被排除在以日光为能量来源的能量利用途径之外.本文介绍了一种新的微生物能量利用途径,即非光能营养微生物通过半导体矿物光催化作用来利用太阳能进行生长.实验室模拟体系中,金属氧化物、金属硫化物等天然半导体矿物在模拟日光激发下产生的光电子促进了化能自养与异养微生物的生长.研究结果表明微生物的生长与光子能量和光子数量密切相关,同时不同波长光辐照下的微生物生长情况与矿物的光吸收谱相吻合.这一能量利用途径的光能-生物能转化效率为0.13‰-1.90‰.在含有天然半导体矿物与天然微生物的红壤体系中,进一步发现半导体矿物光催化能够明显改变环境微生物的种群结构.已有的研究揭示了一种新发现并极有可能长期在地球上存在的微生物能量利用途径,即通过自然界中半导体矿物日光催化作用产生的光电子能够促进非光能营养微生物的生长代谢活动.     

Apple Sucrose Transporter SUT1 and Sorbitol Transporter SOT6 Interact with Cytochrome b5 to Regulate Their Affinity for Substrate Sugars

Sugar transporters are central machineries to mediate cross-membrane transport of sugars into the cells, and sugar availability may serve as a signal to regulate the sugar transporters. However, the mechanisms of sugar transport regulation by signal sugar availability remain unclear in plant and animal cells. Here, we report that a sucrose transporter, MdSUT1, and a sorbitol transporter, MdSOT6, both localized to plasma membrane, were identified from apple (Malus domestica) fruit. Using a combination of the split-ubiquitin yeast two-hybrid, immunocoprecipitation, and bimolecular fluorescence complementation assays, the two distinct sugar transporters were shown to interact physically with an apple endoplasmic reticulum-anchored cytochrome b5 MdCYB5 in vitro and in vivo. In the yeast systems, the two different interaction complexes function to up-regulate the affinity of the sugar transporters, allowing cells to adapt to sugar starvation. An Arabidopsis (Arabidopsis thaliana) homolog of MdCYB5, AtCYB5-A, also interacts with the two sugar transporters and functions similarly. The point mutations leucine-73 → proline in MdSUT1 and leucine-117 → proline in MdSOT6, disrupting the bimolecular interactions but without significantly affecting the transporter activities, abolish the stimulating effects of the sugar transporter-cytochrome b5 complex on the affinity of the sugar transporters. However, the yeast (Saccharomyces cerevisiae) cytochrome b5 ScCYB5, an additional interacting partner of the two plant sugar transporters, has no function in the regulation of the sugar transporters, indicating that the observed biological functions in the yeast systems are specific to plant cytochrome b5s. These findings suggest a novel mechanism by which the plant cells tailor sugar uptake to the surrounding sugar availability.     

Phosphorylation of cardiac troponin I by mammalian sterile 20-like kinase 1

Mst1 (mammalian sterile 20-like kinase 1) is a ubiquitously expressed serine/threonine kinase and its activation in the heart causes cardiomyocyte apoptosis and dilated cardiomyopathy. Its myocardial substrates, however, remain unknown. In a yeast two-hybrid screen of a human heart cDNA library with a dominant-negative Mst1 (K59R) mutant used as bait, cTn [cardiac Tn (troponin)] I was identified as an Mst1-interacting protein. The interaction of cTnI with Mst1 was confirmed by co-immunoprecipitation in both co-transfected HEK-293 cells (human embryonic kidney cells) and native cardiomyocytes, in which cTnI interacted with full-length Mst1, but not with its N-terminal kinase fragment. in vitro phosphorylation assays demonstrated that cTnI is a sensitive substrate for Mst1. In contrast, cTnT was phosphorylated by Mst1 only when it was incorporated into the Tn complex. MS analysis indicated that Mst1 phosphorylates cTnI at Thr(31), Thr(51), Thr(129) and Thr(143). Substitution of Thr(31) with an alanine residue reduced Mst1-mediated cTnI phosphorylation by 90%, whereas replacement of Thr(51), Thr(129) or Thr(143) with alanine residues reduced Mst1-catalysed cTnI phosphorylation by approx. 60%, suggesting that Thr(31) is a preferential phosphorylation site for Mst1. Furthermore, treatment of cardiomyocytes with hydrogen peroxide rapidly induced Mst1-dependent phosphorylation of cTnI at Thr(31). Protein epitope analysis and binding assays showed that Mst1-mediated phosphorylation modulates the molecular conformation of cTnI and its binding affinity to TnT and TnC, thus indicating functional significances. The results of the present study suggest that Mst1 is a novel mediator of cTnI phosphorylation in the heart and may contribute to the modulation of myofilament function under a variety of physiological and pathophysiological conditions.