脂肪酶催化选择性开环聚合反应研究进展

对脂肪酶催化选择性开环聚合反应的研究历史和最新研究进展等进行综述。重点介绍在立体选择性、化学选择性和区位选择性开环聚合反应方面的研究进展,并对该技术的发展趋势和潜在应用进行了展望。     

Metal-ion catalyzed polymerization in the eutectic phase in water-ice: a possible approach to template-directed RNA polymerization

The emergence of an RNA world requires among other processes the non-enzymatic, template-directed replication of genetic polymers such as RNA or related nucleic acids, possibly catalyzed by metal-ions. The absence of uridilate derivative polymerization on adenine containing templates has been the main issue preventing an efficient template-directed RNA polymerization. We report here the investigation of template-directed RNA polymerization in the eutectic phase in water–ice. In particular, it was found that activated uridilate monomers in the presence of metal-ion catalysts could efficiently elongate RNA hairpins whose 5′-overhangs served as the templating sequence. The same applies for every other pyrimidine and purine nucleobase. Moreover, the initial elongation rates were always higher in the presence of a template complementary to the nucleotide than in systems without proper base-pairing opportunities. These results suggest that a template-directed RNA polymerization catalyzed by metal-ions could be carried out under eutectic phase in water–ice conditions.     

Asymmetric synthesis polymerization of meso oxiranes and thiiranes

The asymmetric synthesis polymerization or “enantiogenic” polymerization of some meso oxiranes, cis-dimethyloxirane (cis-DMO) and cyclohexene oxide (CHO), and thiiranes, cis-dimethylthiirane (cis-DMT) and cyclohexene sulfide (CHS), initiated with different chiral systems was examined. Strong differences in behaviour were observed between oxiranes and thiiranes depending on the initiator used. The initiators based on ZnEt₂ or CdMe₂ and a chiral diol give optically active polymers from meso thiiranes but fail to induce an asymmetric polymer synthesis with meso oxiranes. A new chiral initiator based on ZnEt₂ and (1S,2R)-ephedrine allowed us to prepare optically active poly CHOs, which can be fractionated into fractions exhibiting opposite optical activities. © 1992 Wiley-Liss, Inc.     

Helix-sense-selective polymerization of diphenyl-2-pyridylmethyl methacrylate with chiral anionic initiators

Polymerization of diphenyl-2-pyridylmethyl methacrylate was carried out with the complexes of organolithium compounds with 22 chiral ligands. Helix-sense-selectivity of the polymerization was largely affected by a slight structural difference of chiral ligands. (+)-(S)-2-(1-Pyrrolidinylmethyl)pyrrolidine was the most effective ligand in producing a one-handed helical polymer with narrow molecular weight distribution.     

pH敏感的荧光探针及在活细胞中的应用

细胞内的pH是细胞内多种酶活性和生理活动的重要调节因素,准确、动态的监测细胞内pH变化对研究细胞内的活动至关重要。一些荧光小分子可以感应pH的变化,同时具有较高的灵敏度和特异性,对细胞损伤较小且标记操作简单,已逐渐发展成为一种监测细胞内pH变化的有效方法。本文主要介绍目前常用pH敏感的荧光探针及其在活细胞研究中的进展。     

Microbial synthesis of semiconductor CdS nanoparticles,their characterization,and their use in the fabrication of an ideal diode

Cadmium sulfide nanoparticles were synthesized intracellularly by a Schizosaccharomyces pombe strain when challenged with 1 mM cadmium in solution. The nanoparticles, a known semiconducting material, exhibited an absorbance maximum at 305 nm. X-ray scattering data showed that the nanoparticles had a Wurtzite (Cd(16)S(20))-type hexagonal lattice structure and most of the nanoparicles were in the size range of 1-1.5 nm. The nanoparticles were used in the fabrication of a heterojunction with poly (p-phenylenevinylene). The diode exhibited approximately 75 mA/cm(2) current at 10 V when forward biased and the breakdown occurred at approximately 15 V in the reverse biased mode. These characteristics are considered ideal for a diode.     

Preparation of cadmium sulfide nanoparticles and their application for improving the properties of the electrochemical sensor for the determination of enrofloxacin in real samples

An advanced electrochemical sensor for the detection of enrofloxacin (ENR) based on the use of a modified electrode containing cadmium sulfide (CdS) nanoparticles (NPs) is reported. The CdS NPs were synthesized and characterized and then coated onto the electrode to fabricate a modified electrode that exhibited a lower limit of detection of 9.5 × 10^?8 mol·L^?1. This detection limit compares with a traditional electrode that exhibited a concentration detection range of 1.0 × 10^?2 to 1.0 × 10^?7 mol·L^?1. This modified electrode demonstrated good selectivity, reproducibility, response time (<40 s), lifetime (up to 12 wk), and pH range (3.3‐7.2) for the determination of ENR in real samples (eg, pig urine).     

Kinetics of phenolic polymerization catalyzed by peroxidase in organic media

Phenolic polymerization was carried out by enzymatic catalysis in organic media, and its kinetics was studied by using high-pressure liquid chromatography (HPLC). Phenols and aromatic amines with electron-withdrawing groups could hardly be polymerized by HRP catalysis, but phenols and aromatic amines with electron-donating groups could easily be polymerized. The reaction rate of either the para-substituted substrate or meta-substituted substrate was higher than that of ortho-substituted substrate. When ortho-position of hydroxy group of phenols was occupied by an electron-donating group and if another electron-donating group occupied para-position of hydroxy group, the reaction rate increased. Horseradish peroxidase and lactoperoxidase could easily catalyze the polymerization, but chloroperoxidase and laccase failed to yield polymers. Metallic ions such as Mn(2+), Fe(2+), or Fe(3+), and Cu(2+) could poison horseradish peroxidase to various extents, but ions such as Co(2+), Cd(2+), Zn(2+), and K(+) were not found to inhibit the reaction. (c) 1995 John Wiley & Sons, Inc.     

动物活体内细胞光操控的研究进展

动物活体环境下单细胞的光操控对于研究细胞的结构和功能,细胞与组织之间的相互作用,以及细胞病变机理、血栓形成机制和肿瘤细胞迁移等生物医学问题具有重要意义。2013年,光镊技术首次应用于活体动物内单细胞的捕获和操控,开辟了活体动物内光学操控新领域。本文就该领域涉及的活体操控技术及近来取得的重要研究进展进行概述,简要分析了实现深度组织内细胞操控所遇到的技术瓶颈并讨论了解决方案。     

Cysteine-independent polymerization of metallothioneins in solutions and in crystals

Polymerization of metallothioneins is one of the usually encountered puzzles during the research process of metallothioneins' structure and function. Our work focuses on the cysteine independently occurred polymerization from metallothioneins monomers in different milieus, while it leaves out the aggregation caused by the oxidation of cysteine, because the latter circumstance is the result of purification lapsus. After the purification of metallothioneins monomers, a dynamic light-scattering technique is used to detect the polymerized states of rabbit liver metallothionein I and II in different buffers, which is the first systematical detection of polymerized states of metallothioneins in solutions. The effects of different compositions of each buffer are discussed in details. Steric complementarity, hydrophobic, and electrostatic interaction characteristics are studied, following the modeling of monomers and relevant polymers of rat metallothionein II, rabbit liver metallothionein I and II. These theoretical calculations are the first complete computer simulations on different factors affecting metallothioneins' polymerization. A molecular recognition mechanism of metallothioneins' polymerization in solutions is proposed on the bases of experimental results and theoretical calculations. Preliminary X-ray studies of two crystal forms of rabbit liver metallothionein II are compared with the crystal structure of rat metallothionein II, and the polymerized states in crystal packing are discussed with the knowledge of polymerization of metallothioneins in solutions. The hypothesis, which is consistent with theoretical calculations and experimental results, is expected to construct a connection between the biochemical characteristics and physiological functions of metallothioneins, and this research may give some enlightenment to the topics of protein polymerizations.     

Lipase-catalyzed polymerization of lactones and linear hydroxyesters

Ring-opening polymerization of six lactones and condensation polymerization of six linear hydroxyesters were carried out at 45°C using a lipase from Pseudomonas sp. Ring-opening polymerization of the lactones gave both higher molecular weight and higher monomer conversion than condensation of corresponding linear hydroxyesters. Lipase-catalyzed copolymerization of -caprolactone with lactide and cyclopentadecanolide gave higher molecular weight compared with copolymerization of -caprolactone with corresponding linear hydroxyesters. © Rapid Science Ltd. 1998     

准噶尔盆地荒漠生物类群与环境的关系

对荒漠生态系统的研究,近期在我国乃至全球均极为活跃。由于开发的需要和全球沙漠化威胁日趋严重,引起有关人士的关注。就新疆而言,解放前虽有一些学者进行过研究,但仅限于某些方面,直到50年代后期展开大规模综合考察以来,对新疆的荒漠自然条件才有较全面认识。作者近年的研究过程,注意到此区生物类群与环境存在较特殊关系,     

关于生命系统熵势函数的建立及应用

依据非平衡非线性系统理论的广义势函数,建立了可描述生命系统的熵势及其表达式．作为应用,分析了生命系统的相变和生命机体内部的熵力．     

单个活细胞中生物分子动态行为的FRET实时检测

分别采用两种不同绿色荧光蛋白(green fluorescent prote in,GFP)突变体作为荧光共振能量转移(fluo-rescence resonance energy transfer,FRET)对的供体和受体,并利用分子生物学技术将供体和受体分子分别与特定的生物分子融合,这种技术已经成为在单个活细胞中实时长时间检测蛋白质间的动态相互作用的主要技术。主要介绍了基于GFPs的FRET技术在单个活细胞中实时长时间研究生物分子动态行为的应用。     

Molecular mechanisms of the polymerization of fibrin and the formation of its three-dimensional network

The results of biochemical, immunochemical, and X-ray studies of the structures of fibrinogen and fibrin molecules were analyzed. The mechanisms of the successive formation of the fibrin three-dimensional network were described: the polymerization of monomeric molecules with the formation of bifilar protofibrils, the lateral association of protofibrils, and the embranchment of the forming fibrils. Data on the electron and confocal microscopy of the polymeric fibrin were considered. The role of the known polymerization centers of fibrin which participated in the formation of protofibrils and their lateral association was discussed. Data on the existence of the previously unknown polymerization centers were given. In particular, the experimental results demonstrated that one of such centers which participated in the formation of protofibrils was located in the Bβ12–46 fragment, and did not require the cleavage of fibrinopeptide B for its functioning. The results of the computer modeling of the spatial structure of the fibrin(ogen) molecule and the intermolecular interactions in the course of the fibrin polymerization were presented. The location of the αC domains in the fibrin(ogen) molecule and their role in the polymerization process were discussed. Information on the structure of the calciumbinding sites of fibrin(ogen) and the functional role of Ca²⁺ in fibrin polymerization was published. The structure of factor XIII(a) and the mechanisms of fibrin stabilization by this factor were briefly described.     

Immobilized lipase on porous silica particles: Preparation and application for biodegradable polymer syntheses in ionic liquid at higher temperature

Porous silica particles (PSP) modified with different surface active groups were prepared for covalent immobilization of porcine pancreas lipase (PPL). Organosilanes combined with reactive end amino-group or epoxy-group were employed for the modification through silanization process. Polyethylenimine and long chain alkyl silane coupling agent were also used in the modification process. Several modification-immobilization strategies were performed, while good coupling yield could be achieved within the range of 86.2–158.2 mg of native PPL per gram of the carrier. Furthermore, at higher temperature, the resulting immobilized PPL (IPPL) could successfully perform the syntheses of polycaprolactone (PCL) and poly(5,5-dimethyl-1,3-dioxan-2-one) (PDTC) in ionic liquid medium. No polymers could be obtained catalyzed by native PPL, suggesting that IPPL showed much higher catalytic activity than native PPL. Effect of different treatments on the activity of IPPL also showed the long time high temperature stability in ionic liquid medium, contributing to a good combination of immobilization and ionic liquids effect. The catalytic activity of IPPL for polymerization was closely related to both the properties of immobilized enzyme and cyclic monomer. This work would be expected to highlight further careful design of immobilized enzyme for a wide range of application, especially in biodegradable polymers syntheses.     

Geogenically Acidified Mining Lakes — Living Conditions and Possibilities of Restoration

Pyrite and marcasite oxidation in the consequence of lignite surface mining creates lakes with pH as low as 2 to 3, buffered by high contents of iron and aluminium. Living conditions in this extreme habitat for plants and animals are described as well as the characteristics of the pioneer settlement. The utilization of these fish-free lakes is very limited. As possibilities for a water quality improvement special recultivation methods of overburden, chemical neutralization and biological ecotechnologies are recommended.     

Estimation of polymerization efficiency in the formation of polyacrylamide gel,using continuous optical scanning during polymerization

The Ferguson plot and ‘quantitative’ gel electrophoresis (based on the Ferguson plot) depend on a knowledge of accurate gel concentrations. The easiest way to estimate accuracy of gel concentrations, in terms of the degree of completion of the polymerization reaction which gives rise to a gel, is by spectrophotometry. Making use of the apparatus for continuous optical scanning of polyacrylamide gels, the extent and rate of polymerization of cross-linked polyacrylamide were estimated by measuring the absorbance at 275 mm of the reaction mixture subsequent to free radical initiation of polymerization. Under appropriate conditions of monomer concentration, initiator levels and temperature, absorbance decreased monotonically after alag period of 10 min, and after 20–30 min of reaction the absorbance reached a plateau value which provided a measure of polymerization efficiency. Application of a standard curve of absorbance vs. monomer concentration allowed one to quantitate concentrations of residual monomer throughout the course of polymerization. Under a set of arbitrary polymerization conditions (e.g. 6–20% total gel concentration), the reaction went to 63–96% completion. The rate of polymerization was approximately proportional to the square of the monomer concentration (2nd-order reaction kinetics). Absorbance decrease subsequent to the initiation of the polymerization reaction appeared suitable as a measure of efficiency of polymerization since: (1) absorbance spectra of monomers at 0.5%T and residual monomers in a 10%T gel, at a time when polymerization seemed terminated, coincided; (b) values of residual monomer obtained were reasonable (10–30%); (c) bimolecular reaction kinetics were found, in agreement with expectation; and (c) absorbance of incomplete polymerization mixtures, deficient in either initiators or monomers, was constant with time.     

Conformational and functional studies of three gelsolin subdomain-1 synthetic peptides and their implication in actin polymerization

Gelsolin, a calcium and inositol phospholipid-sensitive protein, regulates actin filament length. Its activity is complex (capping, severing, etc.) and is supported by several functional domains. The N-terminal domain alone (S1), in particular, is able to impede actin polymerization. Our investigations were attempted to precise this inhibitory process by using synthetic peptides as models mimicking gelsolin S1 activity. Three peptides issued from S1 and located in gelsolin—actin interfaces were synthesized. The peptides (15–28, 42–55, and 96–114 sequences) were tested for their conformational and actin binding properties. Although the three peptides interact well with actin, only peptide 42–55 affects actin polymerization. A detailed kinetic study shows that the latter peptide essentially inhibits the nucleation step during actin polymerization. In conclusion, the present work shows that the binding of a synthetic peptide to a small sequence located outside the actin—actin interface is essential in the actin polymerization process. © 1997 John Wiley & Sons, Inc. Biopoly 41: 647–655, 1997