海因酶的催化特性及其反应机理*

目前,已发现多种水解海因类的酰胺酶,如D-海因酶、二氢嘧啶酶、尿囊素酶、亚酰胺酶等海因酶。海因酶具有底物依赖的对映体选择性。在金属离子的参与下,它催化海因、5’-单替代海因及其衍生物发生水解。海因酶是有重要应用价值的工业酶,被广泛应用于生产具光学活性的D-和L-型氨基酸。综述了海因酶的催化特性、反应机理及应用前景。     

海因酶研究新进展

海因酶在生产手性药物中间体D-氨基酸上具有广泛的应用价值。全面综述了海因酶的研究历史、分类、进化和酶学性质;总结了产海因酶的茵种筛选和产酶条件、D-海因酶的纯化、微生物海因酶基因的克隆及其在大肠杆茵中的表达等技术;阐述了酶法合成D-(-)-对羟基苯甘氨酸的工艺条件。     

海因酶热稳定性及底物特异性研究进展

海因酶是在微生物中广泛分布的能水解5-取代海因衍生物制备光学纯氨基酸的关键生物催化剂,在各种氨基酸的酶法生产中具有良好的应用前景。着重概述了海因酶的热稳定性、底物特异性研究及应用,并讨论了其发展方向。     

L-海因酶与D-海因酶的序列及结构比较研究

运用生物信息学的研究方法,从序列及结构上对L型及D型海因酶进行了初步的比较。研究了两种类型的海因酶在序列、骨架结构及活性中心的区别,并探讨了产生这些差异的理论基础,为海因酶进一步的理论及应用研究提供一定的指导。     

嗜热菌Bacillus fordii 3-2海因酶的纯化及性质

对自行筛选的海因酶法L-苯丙氨酸生产菌株Bacillus fordii 3-2中的海因酶进行了分离纯化及相关性质研究.该海因酶协同L-氨甲酰水解酶是海因酶法生产L-氨基酸的关键酶.B.fordii 3-2菌悬液经压力破碎离心后取上清液为粗酶液,粗酶液通过硫酸铵分级沉淀、Phenyl FF(high sub) 疏水层析以及Source 15Q离子交换层析,经SDS-PAGE分析达到电泳纯,亚基相对分子质量为55×103,海因酶的纯化回收率为20.5%,纯化倍数为149.23.该海因酶在pH 8.0～10.0的范围内具有较高的活性,在45～70℃具有很高的酶活力, 最适反应pH和温度分别为10.0 ℃和65 ℃.纯酶易氧化失活,DTT对该酶有一定的保护作用.二价金属离子,Mn2 、Co2 及Fe2 对酶活性有显著的促进作用,而Ca2 、Cu2 等对酶活性有抑制作用.相关研究可为该菌株及海因酶的进一步开发与应用提供理论指导.     

假单胞菌海因酶基因在大肠杆菌中的高效表达(英文)

为实现利用生物酶转化法进行D 对羟基苯甘氨酸的工业化生产 ,构建了 3株海因酶基因工程菌 .利用PCR技术从恶臭假单胞菌 (Pseudomonasputida)CPU 980 1染色体DNA中扩增得到长约1.8kb的含编码区和自身启动子的海因酶全基因 .通过将海因酶全基因插入pMD18 T质粒、海因酶基因的编码区与pET 17 b质粒重组、海因酶基因编码区和T7强启动子一起插入pMD18 T质粒分别得到重组质粒pMD dht、pET dht和pMD T7 dht.将上述重组质粒分别转化大肠杆菌 (Escherichiacoli) ,通过地高辛标记菌落原位杂交和海因酶活力测定两种方法 ,筛选出具有海因酶活力的阳性转化子 .结果表明 ,大肠杆菌的RNA聚合酶能够识别和结合来自恶臭假单胞菌海因酶基因的自身启动子 ,该启动子在大肠杆菌中能够工作 .基因工程菌E .coliBL2 1 pMD dht、E .coliBL2 1 pET dht和E .coliBL2 1 pMD T7 dht的海因酶活力分别为 170 0U L、190 0U L和 2 5 0 0U L ,比野生菌P .putidaCPU 980 1的海因酶活力分别提高了 8倍、9倍和 12倍 .薄层扫描结果显示 ,这些工程菌的海因酶表达量分别约占菌体总可溶性蛋白质的 2 0 %、31%和 5 7%.SDS PAGE显示 ,海因酶的单体分子量约为 5 0kD .经工程菌E .coliBL2 1 pMD T7 dht催化 ,底物对羟基苯海因的转化率在 13h内可达到 9     

Eupergit C250L固定化D-海因酶及其催化性质

D-海因酶是海因酶法制备D-氨基酸的关键酶。利用Burkholderic cepecia1003菌发酵产酶,所得海因酶纯化后,以Eupergit C250L为载体进行共价固定化。分别考察了酶液蛋白浓度、固定化时间对蛋白固定量和酶活回收率的影响以及固定化前后海因酶催化性质的变化。结果表明:较高的酶液蛋白浓度和较长的固定化时间均有助于改善海因酶的固定化效果;固定化可显著提高海因酶的最适作用温度,但对其最适作用pH影响不大;固定化后海因酶对D,L-BH和MH的米氏常数均有较大幅度的降低。固定化酶反应器的实验表明:40℃下,底物(D,L-BH)1.0 g.L-1,体积流速1.0 mL.min-1,经21 h转化,产物N-Phe质量浓度可达0.47 g.L-1,转化率达43.21%。     

海因酶与氨甲酰水解酶产生菌的鉴定及分布

利用Biolog微生物鉴定系统和16S rDNA序列分析相结合的方法对自行筛选的24株海因酶和氨甲酰水解酶产生菌进行了鉴定。海因酶与氨甲酰水解酶产生菌主要分布在Bacillus,Geobacillus,Brevibacillus,Aneurinibacillus,Microbacterium,Pseudomonas,Kurthia和Empedobacter等菌属,特别的是,Kurthia和Empedobacter是新的海因酶和氨甲酰水解酶产生菌属,说明海因酶和氨甲酰水解酶产生菌在细菌中分布较广泛。进一步分析比较发现,D-海因酶产生菌主要分布于Pseudomonas,Agrobacterium等菌属中,而大部分L-海因酶产生菌分布于Bacillus,Geobacillus,Microbacterium等菌属中,说明D-海因酶产生菌与L-海因酶产生菌分布特点具有一定的种属倾向性。这些工作不仅提供了多种海因酶和N-氨甲酰水解酶的生物材料,而且对双酶的结构与功能及分子进化研究等具有重要意义。     

海因酶的研究进展

海历酶在生产手性药物中间体上具有广泛的应用价值,本文综述了海因酶的研究历史、分类与进化、酶学性质、工业应用及发展方向。     

聚苯乙烯树脂固定化D-海因酶的初步研究

D-海因酶广泛用于D-氨基酸的制备研究和生产中,目前已有许多固定化D-海因酶及含D-海因酶细胞的研究报道。尝试了不同功能基团的聚苯乙烯树脂进行D-海因酶的固定化,结果表明功能基为伯氨基和仲氨基效果较好,并选取聚苯乙烯树脂D92进行了固定化D-海因酶的研究。采用该树脂制备固定化酶的最优条件是：酶质量浓度6mg／mL、温度25℃、固化时间12h。所得固定化酶的最适作用温度45℃,最适作用pH为8.5,且作用温度及适宜pH较广,Km为游离酶的1,8倍,且储存稳定性、操作稳定性较好。45℃下半衰期为11d。     

略论棕榈科与新分出的省藤科的系统分类、演化、区系地理及主要的经济用途

棕榈科原省藤亚科因其子房壁及外果皮被倒生、螺旋状排列的鳞片所覆盖,而区别于其他亚科,因而独立分出成一新科--省藤科。作者讨论了棕榈科的祖先种可能在石炭纪时,自原始裸子植物开以顿目在分化、衍生出苏铁目祖先种的进化干上,于白垩纪时分化出的一个分支。在棕榈科的祖先种出现不久后,在其进化的分支上,于白垩纪后期又分化出一旁支,成为棕榈科的姊妹科--省藤科的祖先种。从两祖先种分别再分化、衍生出现今分布地球上该二科的属与种。两科、尤其前者是被子植物、尤其是单子叶植物中最原始的类群之一。作者还提出棕榈科象牙椰亚科与贝叶棕亚科是该科最原始或较原始的两类群;槟榔亚科和腊材榈亚科是较进化的两类群;而水椰亚科祖先种可能源于象牙椰亚科的祖先种,但又演化为该科最进化与特化的类群。省藤科省藤亚科略比鳞果榈亚科原始。作者讨论了两科为泛热带分布的科,指出两科的"现代分布区"在南北两半球热带地区,少数种还延伸分布到两半球暖亚热带、甚至达中亚热带地区,分布区边缘最北达日本中部、中国长江流域及黄河下游的南部,美国加利佛尼亚州与佛罗里达州和地中海北部;最南达智利中部和新西兰南部;而"现代分布中心"在热带与暖亚热带的亚洲,中、南美洲,大洋洲及非洲的东、南、西部;但分布区的"密集中心"则在热带亚洲、热带中及南美洲、南太平洋群岛及非洲东南部。作者还介绍了近50年我国南方引种驯化成功的两科植物近400种(见^*图谱),其中少数为耐寒的种类,有的种已引种到长江流域或更北的地区。引种的大部分种都有其重要的经济用途,包括:1. 食用,如淀粉和树液可制"西米"或制糖,酿酒、醋或作饮料;果或种子榨油,供食用或工业用;某些种的嫩芽作蔬菜,甚至种子代咖啡饮用;2. 药用,有消炎、止血、活血、驱虫、抗癌等用;3. 建筑、工艺与日用品,包括不少种的树干供建普通房子、桥梁、小船;少数种可提制工业用蜡;许多种的纤维制高级缆绳和编织品;还制工艺品与日用品等;4. 代表热带景观的园林工程、绿化及美化环境的观赏树和人行道树及建造园林景观生态类型的树种等。     

人类口唇部形态变异的研究进展

口唇部是主要的面部形态区域之一,具有重要的生理功能。嘴唇保护口腔免受外部物质的渗透,维持口腔内部的湿度和温度,帮助咀嚼,同时也是外科美容整形手术中的主要部位之一,且在法医物证研究中唇纹也具有重要的鉴别价值。目前关于口唇部形态的研究主要包括以下几个方面：口唇部形态变异的性别、年龄、族群差异;影响口唇部形态变异的因素,如遗传基因、饮食结构、牙齿及颌骨形态和呼吸方式等;口唇部形态研究在医学、个人身份识别及法医刑侦领域的相关应用等。国内口唇部形态研究相对较少,尤其缺乏较为系统的口唇部形态生长发育及变异研究。本文通过梳理和归纳国内外相关文献的研究数据和结论,对口唇部形态相关研究作简要概述,并对国内口唇部生长发育及形态变异研究作了简要回顾和展望。     

Sugars in crop plants

We review current knowledge of the most abundant sugars, sucrose, maltose, glucose and fructose, in the world's major crop plants. The sucrose‐accumulating crops, sugar beet and sugar cane, are included, but the main focus of the review is potato and the major cereal crops. The production of sucrose in photosynthesis and the inter‐relationships of sucrose, glucose, fructose and other metabolites in primary carbon metabolism are described, as well as the synthesis of starch, fructan and cell wall polysaccharides and the breakdown of starch to produce maltose. The importance of sugars as hormone‐like signalling molecules is discussed, including the role of another sugar, trehalose, and the trehalose biosynthetic pathway. The Maillard reaction, which occurs between reducing sugars and amino acids during thermal processing, is described because of its importance for colour and flavour in cooked foods. This reaction also leads to the formation of potentially harmful compounds, such as acrylamide, and is attracting increasing attention as food producers and regulators seek to reduce the levels of acrylamide in cooked food. Genetic and environmental factors affecting sugar concentrations are described.     

The role of the microbiome in the neurobiology of social behaviour

Microbes colonise all multicellular life, and the gut microbiome has been shown to influence a range of host physiological and behavioural phenotypes. One of the most intriguing and least understood of these influences lies in the domain of the microbiome's interactions with host social behaviour, with new evidence revealing that the gut microbiome makes important contributions to animal sociality. However, little is known about the biological processes through which the microbiome might influence host social behaviour. Here, we synthesise evidence of the gut microbiome's interactions with various aspects of host sociality, including sociability, social cognition, social stress, and autism. We discuss evidence of microbial associations with the most likely physiological mediators of animal social interaction. These include the structure and function of regions of the ‘social' brain (the amygdala, the prefrontal cortex, and the hippocampus) and the regulation of ‘social’ signalling molecules (glucocorticoids including corticosterone and cortisol, sex hormones including testosterone, oestrogens, and progestogens, neuropeptide hormones such as oxytocin and arginine vasopressin, and monoamine neurotransmitters such as serotonin and dopamine). We also discuss microbiome‐associated host genetic and epigenetic processes relevant to social behaviour. We then review research on microbial interactions with olfaction in insects and mammals, which contribute to social signalling and communication. Following these discussions, we examine evidence of microbial associations with emotion and social behaviour in humans, focussing on psychobiotic studies, microbe–depression correlations, early human development, autism, and issues of statistical power, replication, and causality. We analyse how the putative physiological mediators of the microbiome–sociality connection may be investigated, and discuss issues relating to the interpretation of results. We also suggest that other candidate molecules should be studied, insofar as they exert effects on social behaviour and are known to interact with the microbiome. Finally, we consider different models of the sequence of microbial effects on host physiological development, and how these may contribute to host social behaviour.     

云南干热河谷植被与环境研究进展

特殊的地理位置和独特的地形地貌特征组合形成了典型的干热气候环境, 在云南省亚热带高原山地河谷下部发育了一类特有的植被类型, 即干热河谷植被。干热河谷植被具有非地带性和稀有性, 以及由土地利用变化为主的人为活动干扰导致的脆弱性。本文回顾了干热河谷植被的研究历史, 分别从干热河谷的植物群落学和植物区系学、干热河谷植被与土地的关系以及干热河谷植被保护与恢复三个方面进行了总结。植物群落与区系研究主要集中于群落分类、植被分类、群落特征、人为干扰影响、区系特征、性质和起源; 植被与土地关系研究侧重于土壤特性、土地利用/覆盖变化、土地退化及水土流失状况; 植被保护与恢复的热点在植被恢复目标、植被恢复功能区划、植被恢复引种及筛选及植被恢复效益评价研究。未来在这些区域应注重自然灾害及预防、水电工程建设对植被的影响及其响应等方面的研究, 深入开展大尺度植被时空格局的监测和动态服务功能分析。     

Physico-chemical environment in Tjeukemeer with special reference to speciation of algal nutrients

This paper attempts to generalize the major controlling factors and their impacts on the physico-chemical environment in Tjeukemeer in relation to the speciation of algal nutrients.Morphometry, man-made hydrology, climate, chemistry of drainage area and biology of the lake appear to control its physico-chemistry the most. Thus the shallow Tieukemeer is vulnerable to wind and therefore almost always completely mixed and often turbid owing to resuspended sediments. In normal summers the lake is oligohaline, hard, eutrophic and alkaline, and in winter it is hard, eutrophic, alkaline and humus-rich. The implications of these properties on the following physico-chemical components and their relevance for the speciation of algal nutrients are discussed: turbidity, colour, halinity, pH, alkalinity, salinity, conductivity, ionic composition, ionic balance and organic matter.Because of the relationships between the physico-chemical environment on the one hand and the speciation and bio-availability of nutrients on the other hand, special attention is given to the speciation of some major and minor elements as derived from ultra filtration experiments. In this context, the metal binding capacity of the organic matter (mainly fulvic acids) is also considered.     

周期性气候变化与人类适应

从古至今,气候变化特别是周期性气候变化,一直深刻影响着人类社会的变革和发展,从旧石器时代人类起源迁移、新石器时代文化文明演变、历史时期王朝兴衰更替,到工业化以来社会经济发展动荡等,无不留下周期性气候变化影响的烙印。本文依据近年来古气候、古人类、环境考古等研究的新证据、新进展,从周期性气候变化的角度审视人类社会各个发展阶段、关键节点的气候特征;通过典型案例,介绍和分析旧石器、新石器、历史时期不同时空尺度周期性气候变化和人类活动之间复杂的相互作用关系,讨论自然科学和人文社会科学对气候变化与人类活动关系认识的异同,阐述在学科交叉背景下研究气候与人类活动关系的新范式。     

How Romanowsky stains work and why they remain valuable — including a proposed universal Romanowsky staining mechanism and a rational troubleshooting scheme

Abstract

An introduction to the nomenclature and concept of “Romanowsky stains” is followed by a brief account of the dyes involved and especially the crucial role of azure B and of the impurity of most commercial dye lots. Technical features of standardized and traditional Romanowsky stains are outlined, e.g., number and ratio of the acidic and basic dyes used, solvent effects, staining times, and fixation effects. The peculiar advantages of Romanowsky staining are noted, namely, the polychromasia achieved in a technically simple manner with the potential for stain intensification of “the color purple.” Accounts are provided of a variety of physicochemically relevant topics, namely, acidic and basic dyeing, peculiarities of acidic and basic dye mixtures, consequences of differential staining rates of different cell and tissue components and of different dyes, the chemical significance of “the color purple,” the substrate selectivity for purple color formation and its intensification in situ due to a template effect, effects of resin embedding and prior fixation. Based on these physicochemical phenomena, mechanisms for the various Romanowsky staining applications are outlined including for blood, marrow and cytological smears; G-bands of chromosomes; microorganisms and other single-cell entities; and paraffin and resin tissue sections. The common factors involved in these specific mechanisms are pulled together to generate a “universal” generic mechanism for these stains. Certain generic problems of Romanowsky stains are discussed including the instability of solutions of acidic dye–basic dye mixtures, the inherent heterogeneity of polychrome methylene blue, and the resulting problems of standardization. Finally, a rational trouble-shooting scheme is appended.     

农林业系统沙兰杨-小麦、玉米群落类型氮、磷、钾营养元素循环的研究

农林业系统沙兰杨-小麦、玉米群落类型N、P、K营养元素循环研究结果表明:在整个群落中,元素N和P发生损亏,K基本平衡。在植物组分库和枯落物库中,元素N、P、K均发生累积;在土壤库中,元素N、P和K均发生亏损。土壤表层(0—20cm)N的含量是限制农作物正常生长发育的主要环境因子。土壤库中有效N和速效P的含量是限制沙兰杨正常生长发育的主要环境因子。群落的年输入输出量大,枯落物少,营养元素在各组分间的循环量大等是该群落类型营养元素循环的重要特性。     

中国昆虫染色体研究现状与展望

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