超滤技术在生物分离中的应用

“超滤”、“膜分离”和我们日常生活,生产实践息息相关,如安全可用的净化水、血液透析或血液滤过人工肾、发酵产物的分离、牛奶浓缩等。超滤由于成本低．通量高．易于放大的优点,已得到广泛的应用。本文主要介绍了超滤技术在发酵液的澄清（固液分离）、生物产品的精制、药用蛋白的分离纯化以及质粒DNA的分离纯化中的研究进展和应用情况。     

含有赖氨酸溶液的浓缩方法

<正> 申请专利范围:把含有赖氨酸的溶液通过阳、阴离子交换膜之间,进行电透析的精制与浓缩。其特点是使上述含有赖氨酸的溶液温度维持在47或70℃范围内的浓缩方法。发明的详细说明: 本发明是有关含有赖氨酸溶液的高效率精制、浓缩方法。更详细地说明本发明是从含有赖氨酸与其他氨基酸共存的赖氨酸发酵液中,用离子交换膜电透析法高效率有选择性地浓缩赖氨酸的方法。     

应用超过滤膜浓缩发酵液

发酵液的浓缩是从发酵液中提取产品的过程中不可缺少的步骤之一。膜过滤是一种不经蒸发使发酵液直接过滤脱水,以使可溶性物质得到浓缩的过程,整个过程由压力推动,由于膜两面的压力差,迫使水穿过半渗透性膜,可溶性物质得到截留而被浓缩。在膜技术发展的早期阶段,膜通常是由醋酸纤维或聚胺组成。由于膜的容量小,低分子量的有机化合物截留量低,以及膜的清洗困难等因素,还不能大规模地应用。     

有机膜在谷氨酸发酵液分离中的应用研究

目的：采用有机微滤膜和超滤膜连续过滤谷氨酸发酵液,去除菌体蛋白,以利于后续的提取操作。方法：利用微滤膜去除菌体及大分子蛋白等杂质,微滤透过液进入超滤膜系统,进一步去除小分子蛋白及色素等,再利用浓缩连续等电法进行提取,得到谷氨酸。结果：发酵液经过滤后,可溶性蛋白、色素去除率分别可达到86.7%和63.2%,谷氨酸的损失率仅为0.6%;谷氨酸的提取收率和纯度分别可达到95%和99%。结论：利用有机膜系统处理谷氨酸发酵液,可高效去除发酵液中的菌体蛋白和色素等,明显地提高了谷氨酸的提取收率及纯度。     

纳滤技术在生物分离中的应用

纳滤技术近年来已成为生物分离领域中的研究热点之一。本文分别就纳滤技术在食品、医药等方面的应用及其膜污染控制进行较为详细的介绍,综述了近年来纳滤生物分离技术的研究与应用进展。     

浊度法测定发酵液中L－苯丙氨酸含量

本文介绍浊度测定发酵液中Ｌ－苯丙氨酸含量的方法。根据指示菌生长的细胞密度与生长培养基内所含的苯丙氨酸量在一定浓度范围内呈线性关系的原理,摸索了浊度测定的最适条件。结果表明浊度法测定发酵液中的苯丙氨酸含量,简便、准确性高。     

生物物理新技术新方法在LB 膜结构与功能研究中的应用

现代生物物理技术与方法的发展为LB膜科学的基础研究提供了丰富的手段,使单分子膜科学取得了令人瞩目的进步。文章就单分子膜研究领域中物理新技术与新方法如LB膜仪、表面粘度计、X射线与中子散射技术、布鲁斯特角光学显微镜、电子显徽镜、原子力显微镜等的基本实验原理及其在单分子膜物理特性与功能研究中的应用作简单介绍;并鉴于各种仪器的优缺点,作者提出了多仪器联合对单分子膜进行研究的方案和前景进行了探讨。     

厌氧膜生物反应器及其应用研究实例

厌氧膜生物反应器(anaerobic membrane bioreactor,AnMBR)是一种耦合了厌氧发酵和膜工艺的新型废水生物处理技术。其优点包括出水水质优良、有机负荷高、污泥产率低以及生物甲烷回收等。归纳了厌氧发酵的原理,常见厌氧膜生物反应器的结构和种类,以及厌氧膜生物反应器的发展历程及其应用。详细介绍了笔者的最新研究成果:厌氧膜生物反应器在高浓度厨余垃圾处理中的应用,及其在城市污水处理中的研究进展。最后总结了该工艺的应用及发展现状,指出其在高浓度有机废水处理中拥有广阔的应用前景。     

细菌显微追踪技术在生物被膜中的应用

细菌生物被膜是粘附于物体表面的由细菌细胞及其胞外物质组成的复杂膜样物聚集体,具有很强的耐药性和免疫逃逸能力。生物被膜内细菌的代谢活性、运动状态等与浮游细菌有明显区别。近年来,先进的显微成像技术结合新型图像处理方法,在研究细菌的运动、生理等方面发挥了重要作用。本文围绕生物被膜,概述了细菌显微追踪技术在其研究中的应用。主要从细菌的运动方式和生物被膜形成过程的调控两方面出发,介绍了在单细胞水平上利用该技术研究生物被膜的进展,包括细菌的游泳、蹭行、群集运动和多种信号通路调控下生物被膜的形成过程等,并展望了该技术在生物被膜其他相关研究领域的应用前景。     

浊度法测定发酵液中L—苯丙氨酸含量

本介绍浊度测定发酵液中Ｌ－苯丙氨酸含量的方法,根据指示菌生长的细胞密度与生长培养基内所含的苯丙氨酸量在一定浓度范围内呈线性关系的原理,摸索了浊度最和达条件,结果表明浊度法测定发酵液中的苯丙氨酸含量,简便,准确性高。     

Probing the Huntingtin 1-17 Membrane Anchor on a Phospholipid Bilayer by Using All-Atom Simulations

Mislocalization and aggregation of the huntingtin protein are related to Huntington’s disease. Its first exon—more specifically the first 17 amino acids (Htt17)—is crucial for the physiological and pathological functions of huntingtin. It regulates huntingtin’s activity through posttranslational modifications and serves as an anchor to membrane-containing organelles of the cell. Recently, structure and orientation of the Htt17 membrane anchor were determined using a combined solution and solid-state NMR approach. This prompted us to refine this model by investigating the dynamics and thermodynamics of this membrane anchor on a POPC bilayer using all-atom, explicit solvent molecular dynamics and Hamiltonian replica exchange. Our simulations are combined with various experimental measurements to generate a high-resolution atomistic model for the huntingtin Htt17 membrane anchor on a POPC bilayer. More precisely, we observe that the single α-helix structure is more stable in the phospholipid membrane than the NMR model obtained in the presence of dodecylphosphocholine detergent micelles. The resulting Htt17 monomer has its hydrophobic plane oriented parallel to the bilayer surface. Our results further unveil the key residues interacting with the membrane in terms of hydrogen bonds, salt-bridges, and nonpolar contributions. We also observe that Htt17 equilibrates at a well-defined insertion depth and that it perturbs the physical properties—order parameter, thickness, and area per lipid—of the bilayer in a manner that could favor its dimerization. Overall, our observations reinforce and refine the NMR measurements on the Htt17 membrane anchor segment of huntingtin that is of fundamental importance to its biological functions.     

Electrostatics of the FeS clusters in respiratory complex I

Respiratory complex I couples the transfer of electrons from NADH to ubiquinone and the translocation of protons across the mitochondrial membrane. A detailed understanding of the midpoint reduction potentials (E_m) of each redox center and the factors which influence those potentials are critical in the elucidation of the mechanism of electron transfer in this enzyme. We present accurate electrostatic interaction energies for the iron-sulfur (FeS) clusters of complex I to facilitate the development of models and the interpretation of experiments in connection to electron transfer (ET) in this enzyme. To calculate redox titration curves for the FeS clusters it is necessary to include interactions between clusters, which in turn can be used to refine E_m values and validate spectroscopic assignments of each cluster. Calculated titration curves for clusters N4, N5, and N6a are discussed. Furthermore, we present some initial findings on the electrostatics of the redox centers of complex I under the influence of externally applied membrane potentials. A means of determining the location of the FeS cofactors within the holo-complex based on electrostatic arguments is proposed. A simple electrostatic model of the protein/membrane system is examined to illustrate the viability of our hypothesis.     

Analysis of the Role of Vaccinia Virus H7 in Virion Membrane Biogenesis with an H7-Deletion Mutant

Essential vaccinia virus genes are often studied with conditional-lethal inducible mutants. Here, we constructed a deletion mutant lacking the essential H7R gene (the ΔH7 mutant) with an H7-expressing cell line. Compared to an inducible H7 mutant, the ΔH7 mutant showed a defect at an earlier step of virion membrane biogenesis, before the development of short crescent-shaped precursors of the viral envelope. Our studies refine the role of H7 in virion membrane biogenesis and highlight the values of analyzing deletion mutants.     

Transmembrane helix-helix interactions: comparative simulations of the glycophorin a dimer

The glycophorin helix dimer is a paradigm for the exploration of helix-helix interactions in integral membrane proteins. Two NMR structures of the dimer are known, one in a detergent micelle and one in a lipid bilayer. Multiple (4 x 50 ns) molecular dynamics simulations starting from each of the two NMR structures, with each structure in either a dodecyl phosphocholine (DPC) micelle or a dimyristoyl phosphatidylcholine (DMPC) bilayer, have been used to explore the conformational dynamics of the helix dimer. Analysis of the helix-helix interaction, mediated by the GxxxG sequence motif, suggests convergence of the simulations to a common model. This is closer to the NMR structure determined in a bilayer than to micelle structure. The stable dimer interface in the final simulation model is characterized by (i) Gly/Gly packing and (ii) Thr/Thr interhelix H-bonds. These results demonstrate the ability of extended molecular dynamics simulations in a lipid bilayer environment to refine membrane protein structures or models derived from experimental data obtained in protein/detergent micelles.     

Site-directed solid-state NMR measurement of a ligand-induced conformational change in the serine bacterial chemoreceptor

The challenging nature of studies of membrane proteins has made it difficult to determine the molecular mechanism of transmembrane signaling. For the bacterial chemoreceptor family, there are crystal structures of the internal and external domains, structural models of the transmembrane domain, and evidence for subtle ligand-induced conformational changes, but the signaling mechanism remains controversial. We have used a novel site-directed solid-state NMR distance measurement approach, using (13)C(19)F REDOR, to measure a ligand-induced change of 1.0 +/- 0.3 A in the distance between helices alpha 1 and alpha 4 of the ligand-binding domain in the intact, membrane-bound serine receptor. This distance change is shown not to be due to motion of the side chain and thus is due to motion of either the alpha 1 or the alpha 4 helix. Additional distance measurements can be used to determine the type of backbone motion and to follow it to the cytoplasm, to test and refine current proposals for the mechanism of transmembrane signaling. This is a promising general method for high-resolution measurements of local structure in intact, membrane-bound proteins.     

In search of a consensus model of the resting state of a voltage-sensing domain

Voltage-sensing domains (VSDs) undergo conformational changes in response to the membrane potential and are the critical structural modules responsible for the activation of voltage-gated channels. Structural information about the key conformational states underlying voltage activation is currently incomplete. Through the use of experimentally determined residue-residue interactions as structural constraints, we determine and refine a model of the Kv channel VSD in the resting conformation. The resulting structural model is in broad agreement with results that originate from various labs using different techniques, indicating the emergence of a consensus for the structural basis of voltage sensing.     

An experimentally constrained computational model of NMDA oscillations in lamprey CPG neurons

Rhythmicity is a characteristic of neural networks responsible for locomotion. In many organisms, activation of N-methyl-D: -aspartate (NMDA) receptors leads to generation of rhythmic locomotor patterns. In addition, single neurons can display intrinsic, NMDA-dependent membrane potential oscillations when pharmacologically isolated from each other by tetrodotoxin (TTX) application. Such NMDA-TTX oscillations have been characterized, for instance, in lamprey locomotor network neurons. Conceptual and computational models have been put forward to explain the appearance and characteristics of these oscillations. Here, we seek to refine the understanding of NMDA-TTX oscillations by combining new experimental evidence with computational modelling. We find that, in contrast to previous computational predictions, the oscillation frequency tends to increase when the NMDA concentration is increased. We develop a new, minimal computational model which can incorporate this new information. This model is further constrained by another new piece of experimental evidence: that regular-looking NMDA-TTX oscillations can be obtained even after voltage-dependent potassium and high-voltage-activated calcium channels have been pharmacologically blocked. Our model conforms to several experimentally derived criteria that we have set up and is robust to parameter changes, as evaluated through sensitivity analysis. We use the model to re-analyze an old NMDA-TTX oscillation model, and suggest an explanation of why it failed to reproduce the new experimental data that we present here.     

Effect of fatty acids on the membrane fluidity of cultured chick dorsal root ganglion measured by fluorescence photobleaching recovery

The effect of fatty acids on the membrane fluidity in tissue cultured chick embryo dorsal root ganglion was studied by fluorescence recovery method. Lateral motion of the lipid was measured by observing the fluorescent probe, 5-(octadecylthiocarbamoylamino) fluorescence, F18. The effective lateral diffusion coefficient of the membrane was around 0.30 X 10(-8) cm2/sec in control cells, 0.42 X 10(-8) cm2/sec in 2-decenoic acid treated cells, and 0.35 X 10(-8) cm2/sec in valeric acid treated cells. From these results it is concluded that effective mobilities of the membrane complex increased about 40% by the external application of 2-decenoic acid, while valeric acid increased it only 12%. From the physiological results that 2-decenoic acid inhibits the Na-channel, it is suggested that this increase in the membrane fluidity might affect the Na-channel.     

PLD基因的基本功能及在植物中的利用研究现状

磷脂酶D(PLD)是一种重要的磷脂水解酶,它广泛分布于各种植物、动物和微生物(细菌、真菌)中。近几年的研究发现它在植物细胞信号转导、膜运输及降解、脂降解、细胞分化和生殖、细胞防御反应中均发挥重要作用。本文从PLD的基本特性、功能、应用等方面综述了其最新研究进展并讨论了其在植物中的应用前景。