多烯脂肪酸对磷脂胆固醇液晶态结构影响机理

用小角Ｘ 射线散射法、３１Ｐ 核磁共振技术和扫描隧道显微镜技术,对多烯脂肪酸多相脂质体的液晶态结构进行了研究。实验结果表明：胆固醇、多烯脂肪酸、非离子表面活性剂均对ＰＥ 脂质体的液晶态结构有明显的影响。在含有高效分散剂的磷酸缓冲溶液中制成的液晶态油酸多相脂质体和亚油酸多相脂质体均由片层六角相和片层立方相组成, 而蓖麻酸多相脂质体由立方六角相组成。     

生物表面活性剂及其应用

生物表面活性剂 (biosurfactant)是表面活性剂家族中的后起之秀 ,它是由微生物所产生的一类具有表面活性作用的物质。它具有减小表面张力、稳定乳化作用、增加泡沫等作用。它的表面活性作用以及对热、p H的稳定性均与化学合成的表面活性剂相当。但它具有一般的化学合成表面活性剂所无法篦美的优点——与环境的兼容性 ,即它没有毒性 ,并可被生物降解 ,因此它们不会对环境造成不利的影响。随着环保意识的不断增强 ,生物表面活性剂正愈来愈受到人们的关注。1　生物表面活性剂的结构特点生物表面活性剂通常是由微生物产生的 ,且多数是由细菌和…     

槐糖脂的生物合成、发酵及分离纯化

槐糖脂是一类由酵母菌代谢生产的糖脂型生物表面活性剂,具有优越的表/界面活性和稳定性,且无毒、无刺激、易被生物降解,在众多领域具有良好的应用前景,被认为是最具潜力替代化学表面活性剂的生物表面活性剂之一。历经50余年的研究,槐糖脂的发酵生产工艺日趋成熟,但产品的分离纯化仍是难点。本文系统地综述了槐糖脂的结构、生物合成途径,重点关注近年来槐糖脂的发酵生产研究现状,以及分离纯化研究现状。并对槐糖脂的发酵生产和分离研究进行了展望,旨在促进槐糖脂的规模化生产与市场化发展。     

表面活性剂对分枝杆菌KR2菌株降解菲的影响

采用同位素示踪方法,从表面活性剂的浓度、离子类型和直链长度三方面研究了表面活性剂对分枝杆菌KR2菌株降解菲的影响。结果表明,表面活性剂的存在不能促进KR2菌对菲的降解;高浓度表面活性剂(≥20mg·L^-1)的存在,使菲的降解出现延迟期,非离子表面活性剂Tween80在低浓度时(≤10mg·L^-1)可以优先作为营养基质被分枝杆菌KR2菌株利用,表面活性剂的离子类型对菲降解的抑制作用的顺序为阳离子表面活性剂TDTMA>阴离子表面活性剂LAS>非离子表面活性剂Tween80,表面活性剂的直链长度对菲降解的影响为直链越短,对微生物的毒性越大,菲降解得越不完全。     

表面活性剂的增溶作用及在土壤中的行为

表面活性剂胶束的存在是导致难溶有机化合物(HOCs)溶解度增加的主要原因,表面活性剂对土壤的影响很大,即使很低浓度的表面活性剂也会明显改变土壤的物理、化学和生物性质,其中表面活性剂的吸附过程起了主要作用,另外,表面活性剂的类型、结构和浓度以及所处环境条件和微生物种类都对土壤中植物、微生物生长和其本身的生物降解和去除有影响,这些都将导致土壤中原有污染物迁移转化的改变,应该引起人们的日益重视。     

生物表面活性剂合成的促产因子研究进展

生物表面活性剂是微生物产生的一类具有表面活性的代谢产物,与化学表面活性剂相比,具有高效、低毒、易降解的优点。本文综述了氨基酸、酵母提取物、金属离子和有机酸作为促产因子时,对生物表面活性剂产量、结构和同系物组成影响的研究进展,并对其促产机理进行了归纳总结,最后对此领域的研究进行了展望。     

酶法合成表面活性剂

用生物技术来研制表面活性剂是当前国际生物工程领域中发展起来的一个新课题。本文概述了用酶法合成五种类型的表面活性剂,单酰化甘油酯类表面活性剂,糖酯类表面活性剂,氨基酸类表面活性剂,磷酯类表面活性剂及氨基酸糖类表面活性剂。     

微生物表面活性剂应用新进展北大核心

表面活性剂是一种重要的工业原料,微生物表面活性剂是表面活性剂的一种,由于其来源于微生物,具有高效且低毒的特点,是一种环境友好型的表面活性剂,逐渐成为近年来表面活性剂研究的热点,并已在多个领域进行了应用尝试。该文从不同类型的微生物表面活性剂入手,阐述近几年来不同类型的微生物表面活性剂在不同领域的应用,比较了不同种类微生物表面活性剂的应用现状,同时对微生物表面活性剂应用存在的问题进行了分析,并对微生物表面活性剂未来的发展趋势进行了展望。     

光及光热纳米材料在生物传感、药物靶向运输和生物成像中的应用

纳米材料是纳米科学技术的重要发展方向之一。纳米材料的结构赋予了其独特的光学性质,纳米尺寸方便其经EPR效应或表面修饰靶向肿瘤组织,并且部分纳米材料可吸收外部光源能量,将其转化为热能。因此,纳米材料在光学传感器、生物成像、药物靶向运输及肿瘤光热治疗中的应用十分广泛。综述主要分类介绍了光学纳米材料和光热纳米材料的优异特性,阐述了其在以上领域中的应用;最后,对纳米材料未来的发展作出了展望。     

产业纵横

表面活性剂改善蛋白质鉴定 ProteasMAX表面活性剂增强了在制备质谱或液相色谱分析用的胰蛋白酶和其它蛋白酶的酶促性能。该表面活性剂使科学家能够在蛋白质分析以前获得更完全的消化,在更短的时间内获得更精确的数据,并且降低样品降解的风险。通过暴露由于二级或三级结构一般接触不到的酶切位点,该产品改善了蛋白质的消化作用;     

Yolk spherocrystal: the structure, composition and liquid crystal template

The structure and composition of the yolk spherocrystal, a biomineral developed in the egg yolk sac during the incubation of a chicken embryo, were investigated through various modern analytical methods. Additionally, inside the yolk sac, yolk liquid crystal, a liquid crystalline phase of lipid developed during the incubation of the embryo, was found and investigated. The spherocrystal was found to be a composite composed of calcium carbonate (vaterite and calcite, primarily the former) and the yolk liquid crystal, which is believed to act as an organic template for spherocrystals mineralization, in a concentric multi-layered sphere structure. Moreover, the yolk liquid crystal was found to have a concentric multi-layered spherical structure and a composition consistent with lecithin. We believed that the spherocrystals function as a reservoir for the storage of calcium in the egg yolk sac during the development of the embryo.     

Protein nucleation and crystallization by homologous protein thin film template

A new method of protein nucleation and crystallization based on Langmuir-Blodgett technology is here utilized for the template stimulation of crystal growth of so far non-crystallized proteins. Microcrystals (60-120 microm) of bovine cytochrome P450scc and human protein kinase CKII alpha subunit were obtained with use of the homologous protein thin film template by vapor diffusion modified hanging drop method. The induction of microcrystals nucleation by the thin template confirms in the two different important classes of proteins, until now never crystallized, the positive stimulatory influence for crystal formation of protein thin film template, which was observed in an earlier study with a model system (chicken egg white lysozyme) as an unexpected acceleration and enhancement in the crystal growth.     

Molecularly imprinted thin film self-assembled on piezoelectric quartz crystal surface by the sol-gel process for protein recognition

A novel method of combining sol-gel and self-assembly technology to prepare a human serum albumin (HSA)-imprinted film on the surface of piezoelectric quartz crystal (PQC) Au-electrode modified with thioglycolic acid was described in this paper. The imprinting process was characterized by using the piezoelectric quartz crystal impedance (PQCI) technique and electrochemical impedance technique. Scanning electron microscope (SEM) was employed to characterize the surface morphology of the resultant imprinted film. The piezoelectric technique and electrochemical impedance technique were also employed to investigate the binding performance of the sol-gel-imprinted film with the template protein. The results showed that the imprinted PQC film can give selective recognition to the template protein. The effects of salts and solvents on the binding capacity of the imprinted film with protein were discussed in detail. Other influencing factors (temperature and pH) have also been investigated. This self-assembly sol-gel imprinting technique was proved to be an alternative method for the preparation of biomacromolecule-imprinted thin film.     

Atomic structure of a CK2alpha human kinase by microfocus diffraction of extra-small microcrystals grown with nanobiofilm template

Extra-small microcrystals of a human kinase CK2alpha were obtained for the first time by the optimization of a recent protein crystallization method based on highly packed protein nanofilm template. Protein crystal induction and growth appear indeed optimal at high surface pressure of the film template yielding high protein orientation and packing. The resulting extra-small CK2alpha microcrystals (of about 20 microm in diameter) was subsequently used for synchrotron radiation diffraction data collection, which proves possible by means of the Microfocus Beamline at the ESRF Synchrotron in Grenoble. The quality of the resulting crystal diffraction patterns and of its resulting atomic structure at 2.4 A resolution proves the unique validity of the above two combined frontier technologies in defining a new approach to structural proteomics capable to solve the atomic structure of proteins so far never been crystallized and of pharmaceutical relevance. Physical explanation in terms of template dipole moments and possibility of generalization of this method to the wide class of proteins not yet crystallized are finally discussed. The structure of our CK2alpha mutant is in the Protein Data Bank (PDB ID Code 1NA7, deposited on 27 November 2002).     

Efficient conformational sampling of local side-chain flexibility

Side-chain flexibility of ligand-binding sites needs to be considered in the rational design of novel inhibitors. We have developed a method to generate conformational ensembles that efficiently sample local side-chain flexibility from a single crystal structure. The rotamer-based approach is tested here for the S1' pocket of human collagenase-1 (MMP-1), which is known to undergo conformational changes in multiple side-chains upon binding of certain inhibitors. First, a raw ensemble consisting of a large number of conformers of the S1' pocket was generated using an exhaustive search of rotamer combinations on a template crystal structure. A combination of principal component analysis and fuzzy clustering was then employed to successfully identify a core ensemble consisting of a low number of representatives from the raw ensemble. The core ensemble contained geometrically diverse conformers of stable nature, as indicated in several cases by a relative energy lower than that of the minimised template crystal structure. Through comparisons with X-ray crystallography and NMR structural data we show that the core ensemble occupied a conformational space similar to that observed under experimental conditions. The synthetic inhibitor RS-104966 is known to induce a conformational change in the side-chains of the S1' pocket of MMP-1 and could not be docked in the template crystal structure. However, the experimental binding mode was reproduced successfully using members of the core ensemble as the docking target, establishing the usefulness of the method in drug design.     

Using a combined transcription-translation system for determining the role of cryptic plasmids of Bacillus thuringiensis

The possibility of entomocyde crystal protein synthesis was studied using a heterological cell-free system with Bacillus thuringiensis plasmid DNA as template. The high level of template activity is usual for Bac. thuringiensis plasmid DNA. Immunochemical studies of the in vitro synthesized polypeptides showed that Bac. thuringiensis plasmid DNA does not direct crystal protein synthesis.     

鸡胚肝脏液晶类脂滴冰冻蚀刻结构研究

鸡胚肝脏组织中液晶类脂滴在不同温度及不同降温速度等条件下,呈现出液晶、结晶及各向同性态,并且三相之间可以互相转变.以冰冻蚀刻复型的方法,证实鸡胚发育中肝脏类滴液晶态为迟晶型液晶,并对0℃处理后的类脂满结晶进行了观察.最后对鸡胚肝脏液晶类脂滴与低密度脂蛋白的相似性进行了讨论.     

A leucine-rich repeat assembly approach for homology modeling of the human TLR5-10 and mouse TLR11-13 ectodomains

So far, 13 groups of mammalian Toll-like receptors (TLRs) have been identified. Most TLRs have been shown to recognize pathogen-associated molecular patterns from a wide range of invading agents and initiate both innate and adaptive immune responses. The TLR ectodomains are composed of varying numbers and types of leucine-rich repeats (LRRs). As the crystal structures are currently missing for most TLR ligand-binding ectodomains, homology modeling enables first predictions of their three-dimensional structures on the basis of the determined crystal structures of TLR ectodomains. However, the quality of the predicted models that are generated from full-length templates can be limited due to low sequence identity between the target and templates. To obtain better templates for modeling, we have developed an LRR template assembly approach. Individual LRR templates that are locally optimal for the target sequence are assembled into multiple templates. This method was validated through the comparison of a predicted model with the crystal structure of mouse TLR3. With this method, we also constructed ectodomain models of human TLR5, TLR6, TLR7, TLR8, TLR9, and TLR10 and mouse TLR11, TLR12, and TLR13 that can be used as first passes for a computational simulation of ligand docking or to design mutation experiments. This template assembly approach can be extended to other repetitive proteins.     

固氮酶CrFe蛋白和MnFe蛋白的空间晶体生长

从分别生长于含Mn和Cr培养基中的棕色固氮菌(Azotobacter vinelandii Lipmann)突变种UW3分离纯化出MnFo和CrFe蛋白.为适应包括固氮酶在内的氧敏感蛋白的空间晶体生长的要求,应用简易而适用的厌氧加样装置代替固氮酶实验室所用的笨重厌氧箱(dry box),在地面进行厌氧加样.在充满氮气的简便有机玻璃箱内厌氧加样的所有样品中,分别用液/液扩散法和汽相扩散的坐滴法都可在一周内使MnFe和CrFe蛋白在宇宙飞船上从溶液中结晶出来.在所用的数种蛋白沉淀剂中,飞船上形成的所有晶体都为单晶,而地面上在多数沉淀剂中部生成大量孪晶.在相同沉淀剂中用液/液扩散法,飞船上生成CrFe蛋白的最大晶体比地面生成的最大晶体大1倍.而在相同沉淀剂中用汽相扩散的坐滴法,飞船上生成的MnFe蛋白最大晶体却没有地面生成的最大晶体大.这种差异也许是由不同结晶方法而不是不同蛋白所引起的.     

DNA adduct bypass polymerization by Sulfolobus solfataricus DNA polymerase Dpo4: analysis and crystal structures of multiple base pair substitution and frameshift products with the adduct 1,N2-ethenoguanine

1,N(2)-Etheno(epsilon)guanine is a mutagenic DNA lesion derived from lipid oxidation products and also from some chemical carcinogens. Gel electrophoretic analysis of the products of primer extension by Sulfolobus solfataricus P2 DNA polymerase IV (Dpo4) indicated preferential incorporation of A opposite 3'-(1,N(2)-epsilon-G)TACT-5', among the four dNTPs tested individually. With the template 3'-(1,N(2)-epsilon-G)CACT-5', both G and A were incorporated. When primer extension was done in the presence of a mixture of all four dNTPs, high pressure liquid chromatography-mass spectrometry analysis of the products indicated that (opposite 3'-(1,N(2)-epsilon-G)CACT-5') the major product was 5'-GTGA-3' and the minor product was 5'-AGTGA-3'. With the template 3'-(1,N(2)-epsilon-G)TACT-5', the following four products were identified by high pressure liquid chromatography-mass spectrometry: 5'-AATGA-3', 5'-ATTGA-3', 5'-ATGA-3', and 5'-TGA-3'. An x-ray crystal structure of Dpo4 was solved (2.1 A) with a primer-template and A placed in the primer to be opposite the 1,N(2)-epsilon-G in the template 3'-(1,N(2)-epsilon-G)TACT 5'. The added A in the primer was paired across the template T with classic Watson-Crick geometry. Similar structures were observed in a ternary Dpo4-DNA-dATP complex and a ternary Dpo4-DNA-ddATP complex, with d(d)ATP opposite the template T. A similar structure was observed with a ddGTP adjacent to the primer and opposite the C next to 1,N(2)-epsilon-G in 3'-(1,N(2)-epsilon-G)CACT-5'. We concluded that Dpo4 uses several mechanisms, including A incorporation opposite 1,N(2)-epsilon-G and also a variation of dNTP-stabilized misalignment, to generate both base pair and frameshift mutations.