嗜极菌的极端酶的若干研究进展

综述了近年来国内外关于嗜极菌的极端酶研究的若干新进展，初步分析了极端酶在极端环境中保持稳定性及活性的结构特性，并介绍了当前极端酶的生物技术进展及其广阔的应用前景。     

在极端条件下具有活力的酶

概述了在极端环境条件下酶的性质,环境对酶的影响,介绍了几种在极端条件下具有活力的酶及其应用。     

极端微生物的功能利用

主要介绍了极端微生物的种类及其功能利用。极端微生物是一些在高温、低温、高压、高盐、强酸、强碱、强辐射等极端环境中生存、繁殖的生命体，具有独特的分子组成、物理化学性能、酶特性及代谢功能等，它们的存在不仅挑战传统生物学理论，而且具有很高的实际应用价值。     

极端微生物在环境保护中的应用

本文介绍了极端微生物的类型及其特点,综述了极端微生物及其产生的极端酶在清洁生产、环保型生物材料的生产及环境污染治理中的应用前景及对环境保护的意义。     

极端微生物在环境保护中的应用

本文介绍了极端微生物的类型及其特点,综述了极端微生物及其产生的极端酶在清洁生产、环保型生物材料的生产及环境污染治理中的应用前景及对环境保护的意义.     

极端微生物的多样性及其应用

极端微生物通常分为五个类群：嗜热微生物、嗜冷微生物、嗜酸微生物、嗜碱微生物、嗜盐微生物．极端环境中的微生物为了适应生存，逐步形成了独特的结构和生理机能，以适应环境．因此，研究适应机理并利用其特殊生理机能具有重要的理论和实际意义，极端微生物能产生多种极端酶和其他生物活性物质，极端微生物资源的开发利用有着广阔的前景．     

极端微生物研究进展

介绍极端微生物的分类学地位和生物类群的概念、发现与分布,综述各种极端微生物的生理机制,阐述各种极端微生物的应用前景和研究意义.     

真核生物DNA聚合酶δ的研究现状

DNA聚合酶δ(DNA polymerase δ)是真核生物DNA复制的主要复制酶,同时还参与DNA修复,对保持真核生物基因组的结构完整性和遗传稳定性具有重要作用.对DNA聚合酶δ蛋白功能活性及其基因表达机制的研究因受技术上的限制而未能深入研究.由于其重要的生物学功能,目前引起人们的更多关注和重视.文中就该酶的生物学功能、亚基组成、核心酶的分子表达调控以及与其他蛋白相互作用等方面对国内外DNA聚合酶δ的研究进行简要综述.     

日本极端民族主义及其教育根源

日本最近的许多政治活动表明了极端民族主义抬头的倾向。日本的极端民族主义有着深刻社会根源和思想根源，但历史上日本政府对教育的指导思想也极大促进了极端民族主义的发展。近代以来的历次教育改革证实了这一点。日本历史教科书事件反映出日本对待历史和历史教育的一贯不诚实态度，正是这种态度为极端民族主义 的诞生和发展提供了温床。极端民族主义是军国主义的前奏，其抬头的倾向足以引起亚洲其他国家的警惕。     

经典力学在极端情况下的应用

给出了经典力学的适用范围，通过计算第3宇宙速度和对原子核半径进行估算，分析了经典力学在极端条件下的应用。所举的2个例子具有一定的代表性，对学习物理学理论、了解物理学思想有一定的意义。     

人工酶与定向进化的前沿与挑战

 定向进化是在实验室环境中，在分子水平上模拟进化过程，得到具有期望特征的蛋白质的方法，目前已成为蛋白质设计改造的重要方法。定向进化不仅可以用于天然蛋白质的改造，也可以通过改造现有的酶，使其具有新的催化活性，从而构建人工酶。本文重点介绍工业生物催化、纳米酶设计和光催化3个方向的前沿成果，并讨论人工酶与定向进化领域存在的挑战和问题。     

Mandelate racemase and muconate lactonizing enzyme are mechanistically distinct and structurally homologous

Mandelate racemase (MR) and muconate lactonizing enzyme (MLE) catalyse separate and mechanistically distinct reactions necessary for the catabolism of aromatic acids by Pseudomonas putida. The X-ray crystal structure of MR, solved at 2.5 A resolution, reveals that the secondary, tertiary and quaternary structures of MR and MLE are remarkably similar; also, MR and MLE are about 26% identical in primary structure. However, MR has no detectable MLE activity and vice versa. Thus, MR and MLE constitute the first example of enzymes that catalyse different reactions, as opposed to mechanistically identical reactions on different substrates, yet possess sufficient structural and sequence identity that they are likely to have evolved from a common ancestor. The discovery that MR and MLE catalyse different reactions but share a common structural framework has broad implications for the natural evolution of enzymes and metabolic pathways, as well as for the rational modification of enzyme activities.     

海藻糖TPS／TPP生物合成途径的进化研究

海藻糖是生物体内一种典型的应激代谢物,在干旱、缺氧等逆境环境下对生物体蛋白质具有特殊的保护作用,大多数生物均采用TPS/TPP途径合成自身所需要的海藻糖。采用生物信息学的方法,对GenBank数据库中收录的TPS/TPP途径中所有TPS和TPP合成酶的蛋白序列进行进化研究,发现该合成途径中的两个海藻糖合成酶具有相当高的进化保守性。     

Selection in vitro of an RNA enzyme that specifically cleaves single-stranded DNA.

The discovery of RNA enzymes has, for the first time, provided a single molecule that has both genetic and catalytic properties. We have devised techniques for the mutation, selection and amplification of catalytic RNA, all of which can be performed rapidly in vitro. Here we describe how these techniques can be integrated and performed repeatedly within a single reaction vessel. This allows evolution experiments to be carried out in response to artificially imposed selection constraints. We worked with the Tetrahymena ribozyme, a self-splicing group I intron derived from the large ribosomal RNA precursor of Tetrahymena thermophila that catalyses sequence-specific phosphoester transfer reactions involving RNA substrates. It consists of 413 nucleotides, and assumes a well-defined secondary and tertiary structure responsible for its catalytic activity. We selected for variant forms of the enzyme that could best react with a DNA substrate. This led to the recovery of a mutant form of the enzyme that cleaves DNA more efficiently than the wild-type enzyme. The selected molecule represents the discovery of the first RNA enzyme known to cleave single-stranded DNA specifically.     

New strategies of protein engineering——directed evolution of enzyme in vitro

New and highly effective strategies for directed enzyme evolution in vitro have been developed in the protein engineering field. They allow engineering all kinds of enzymes in vitro so that new ones with novel functions and features can be obtained by the methods of error-prone PCR, DNA shuffling (exon shuffling), hybrid enzyme, random-priming in vitro recombination (RPR), stagger extension process (StEP), random-directed mutagenesis in vitro, etc., even with little knowlodge of spatial structure and catalytic mechanism of enzymes in advance. The process that would take millions of years in nature can in principle be accomplished in the test within several years.     

Kemp elimination catalysts by computational enzyme design

The design of new enzymes for reactions not catalysed by naturally occurring biocatalysts is a challenge for protein engineering and is a critical test of our understanding of enzyme catalysis. Here we describe the computational design of eight enzymes that use two different catalytic motifs to catalyse the Kemp elimination-a model reaction for proton transfer from carbon-with measured rate enhancements of up to 10(5) and multiple turnovers. Mutational analysis confirms that catalysis depends on the computationally designed active sites, and a high-resolution crystal structure suggests that the designs have close to atomic accuracy. Application of in vitro evolution to enhance the computational designs produced a >200-fold increase in k(cat)/K(m) (k(cat)/K(m) of 2,600 M(-1)s(-1) and k(cat)/k(uncat) of >10(6)). These results demonstrate the power of combining computational protein design with directed evolution for creating new enzymes, and we anticipate the creation of a wide range of useful new catalysts in the future.     

Directed evolution of new catalytic activity using the alpha/beta-barrel scaffold

In biological systems, enzymes catalyse the efficient synthesis of complex molecules under benign conditions, but widespread industrial use of these biocatalysts depends crucially on the development of new enzymes with useful catalytic functions. The evolution of enzymes in biological systems often involves the acquisition of new catalytic or binding properties by an existing protein scaffold. Here we mimic this strategy using the most common fold in enzymes, the alpha/beta-barrel, as the scaffold. By combining an existing binding site for structural elements of phosphoribosylanthranilate with a catalytic template required for isomerase activity, we are able to evolve phosphoribosylanthranilate isomerase activity from the scaffold of indole-3-glycerol-phosphate synthase. We find that targeting the catalytic template for in vitro mutagenesis and recombination, followed by in vivo selection, results in a new phosphoribosylanthranilate isomerase that has catalytic properties similar to those of the natural enzyme, with an even higher specificity constant. Our demonstration of divergent evolution and the widespread occurrence of the alpha/beta-barrel suggest that this scaffold may be a fold of choice for the directed evolution of new biocatalysts.