组成型表达粪产碱杆菌青霉素G酰化酶重组大肠杆菌发酵条件研究

目的:对重组大肠杆菌组成型表达粪产碱杆菌青霉素G酰化酶(AfPGA)进行了发酵条件研究。方法:在摇瓶和5L发酵罐中研究了(NH4)2SO4和葡萄糖浓度对质粒的分离稳定性及青霉素G酰化酶表达的影响。结果:该工程菌质粒具有分离不稳定性,培养基中无(NH4)2SO4时发酵过程中pH和糊精水解生成葡萄糖的浓度变化较小,细胞前期(0h-12h)的生长速率降低,质粒分离稳定性和青霉素G酰化酶的表达水平提高。发酵过程中维持低葡萄糖水平可以限制细胞的生长速率,提高质粒稳定性和促进青霉素G酰化酶的合成。采用混合碳源发酵,发酵培养基含糊精2g/L,12h后以1g/L.h恒速流加葡萄糖至35h,控制流加过程葡萄糖浓度0.1g/L左右,平均比生长速率为0.06h-1,发酵结束时质粒稳定性为86%,青霉素G酰化酶的表达水平达23 000U/L。结论:重组大肠杆菌组成型表达青霉素G酰化酶的研究对工业生产有一定指导意义。     

The Tetracycline Fermentation and Its Regulation

Abstract

The β-glucans different from cellulose are the most abundant class of polysaccharides. They are found in microorganisms and higher plants as structural entities of cell wall, as cytoplasmic and vacuolar reserve materials, and as extracellular substances. Enzyme systems capable to hydrolyze β-glucans are produced by different microorganisms. The occurrence and nature of β-glucanases and their substrates are reviewed. The regulation of biosynthesis of these enzymes, their properties, substrate and product specificities, mode of action and molecular cloning are described. The participation of β-glucanases in the morphogenetic events of yeast cell is presented. The role and synergism of different types of 1,3-β-glucanases in microbial cell wall lysis and the potential application for isolation of intracellular materials like proteins, carbohydrates, enzymes and as an analytical tool are discussed in the light of current knowledge.     

Neuronal Morphology: Shape Characteristics and Models

This paper is focused on quantification (morphometry) and modeling of neuronal morphological complexity. First, computer-aided methods for reconstruction, processing, and analysis of raw morphological data are reviewed. Then, topological and metrical measures are touched upon. Fractal measures (together with the extension of multiscale fractal dimension) are presented more explicitly. Models of neuronal arborizations are differentiated between reconstruction models and growth models (stochastic or mechanistic). The growth model approach is discussed in more detail. The methods presented are applied to several types of neurons and shown to have considerable discriminative power. Recent developments stress the importance of these methods for optimizing virtual neuronal trees in view of functional characteristics of the neurons. Neirofiziologiya/Neurophysiology, Vol. 40, No. 4, pp. 366–372, July–August, 2008.     

Penicillin binding proteins: key players in bacterial cell cycle and drug resistance processes

Bacterial cell division and daughter cell formation are complex mechanisms whose details are orchestrated by at least a dozen different proteins. Penicillin-binding proteins (PBPs), membrane-associated macromolecules which play key roles in the cell wall synthesis process, have been exploited for over 70 years as the targets of the highly successful beta-lactam antibiotics. The increasing incidence of beta-lactam resistant microorganisms, coupled to progress made in genomics, genetics and immunofluorescence microscopy techniques, have encouraged the intensive study of PBPs from a variety of bacterial species. In addition, the recent publication of high-resolution structures of PBPs from pathogenic organisms have shed light on the complex intertwining of drug resistance and cell division processes. In this review, we discuss structural, functional and biological features of such enzymes which, albeit having initially been identified several decades ago, are now being aggressively pursued as highly attractive targets for the development of novel antibiotherapies.     

Conceptual Site Models and Multimedia Modeling: Comparing MEPAS,MMSOILS, and RESRAD

The use of multimedia models to assess current and future human and ecological exposure to contamination at hazardous waste sites has become common practice in recent years. The U.S. Environmental Protection Agency has identified development of a conceptual site model as a critical part of the risk assessment process. This paper investigates the relationship between the choice of conceptual site models and application of multimedia models and the variation in risk estimates obtained when using data and default parameter values suggested by the individual model developers.     

不测土条件下半定量施肥原理和模型评述

1　不测土条件下半定量推荐施肥的需求矿质营养学说是施肥最基本原理 ,它的诞生标志着人类主动施肥时期的开始。从早期的定性施肥阶段 ,发展到今天的定量施肥阶段 ,如何确定施肥量一直是施肥技术的核心和难点。测土是定量施肥或精确施肥的前提。然而 ,测土本身存在各种偏差或精度、测土需要费用 ,本文仅对代表性的不测土施肥模型及其原理进行评述。即使有了土壤有效养分的测定结果 ,如何将其转化为一季内土壤提供的养分量一直是个理论难题 ,特别是Ｎ素土壤供应量更为难测或难以估算。因此 ,如何在使用定量施肥模型的同时 ,而又将土壤养分因…     

Coupling of Fermentation and Esterification: Microbial Esterification of Decanoic Acid with Ethanol Produced via Fermentation

Two different kinds of bioprocess, ethanol fermentation and subsequent microbial esterification, were coupled using Issatchenkia terricola IFO 0933 in an interface bioreactor. The strain produced ethyl decanoate (Et-DA) by esterification of exogenous decanoic acid (DA) with ethanol produced via fermentation. The efficiency of the new coupling system depended on the concentration of glucose in a carrier and DA in an organic phase (decane) in an agar plate interface bioreactor. Optimum glucose content and DA concentration were 4% and 29 mM, respectively.     

抗真菌拮抗放线菌的筛选及摇床发酵条件的优化

从陕西渭北旱原的土壤中分离到 1株对水稻稻瘟、小麦赤霉、棉花枯萎、小麦根腐、小麦全蚀、梨黑星等多种植物病原真菌拮抗性好的放线菌。对其进一步纯化后 ,进行发酵条件的优化 ,认为选用 1 2号培养基 ,pH 8.0 ,培养 72h ,3 0～ 3 4℃ ,1 5 0r/min条件为最佳。     

产L—肉碱的菌种筛选及发酵条件优化

将D,L-肉碱(carnitine,β-羟基-γ-三甲胺丁酸)用浓硫酸脱水获得反-巴豆甜菜碱(trans-crotonobetaine),从本室保藏的菌株中筛选出1株能将反-巴豆甜菜碱非对称合成L-肉碱的菌株E.coliK74.利用它的休止细胞立体选择性地水合反-巴豆甜菜碱产生L-肉碱,起催化作用的酶是L-肉碱脱水酶,是一种可诱导的胞内酶,当培养基中加入反-巴豆甜菜碱并在部分厌氧条件下可诱导产生.如培养基中含有葡萄糖、硝酸盐或氧时,酶的合成受到抑制,在磷酸缓冲液中,E.coliK74休止细胞的最适反应条件是pH为7.8,温度为37～42℃.     

海洋细菌LU—B02生物活性物质发酵条件及理化性质研究

从辽宁渤海水域分离得到的多株细菌中筛选出1株能产生广谱抗菌物质的海洋细菌LU-B02,该菌对白色念珠菌、Y76等酵母状真菌具有较强的抗性,对一些海洋生物体病原菌或寄生菌以及农作物病原性丝状真菌也具有一定的抗性.在此基础上采用均匀设计对该菌株产生生物活性物质的发酵培养基进行选优;比较了不同菌龄、不同发酵时间LU-B02产生生物活性物质的浓度大小,考察了发酵液中生物活性物质对温度、pH值的稳定性;同时对生物活性物质的水溶性、离子特性等性质进行了研究.结果表明LU-B02菌龄16～24h、28℃下发酵26h产生的生物活性物质浓度最高,该物质为强碱性,水溶性强,对温度、pH值较稳定,为该活性物质的进一步提取精制和结构鉴定提供了依据.     

竹黄菌发酵天然色素及其结构的初步研究

对竹黄菌发酵天然色素的条件进行了研究,确定培养基的碳源、氮源、温度和装液量分别为:木糖、硝酸钠、28℃和60mL/250mL。通过利用化学试剂显色反应、紫外光谱(UV)和液质连用(LCMS)等手段初步确定该天然色素为蒽醌类化合物。     

K88大肠杆菌菌毛蛋白发酵工艺条件优化研究

分别采用LB培养基、牛肉膏蛋白胨培养基、强化营养培养基、玉米浆培养基对大肠杆菌K88进行发酵培养,选出最适于大肠杆菌K88生长的玉米浆培养基;采用正交实验对玉米浆培养基的C／N、K2HPO4／KH2PO4、Mg^2＋的配比进行优化,筛选最适于大肠杆菌K88生长的营养配比;研究生长曲线、接种量以及菌体和菌毛生产量的相关性,根据实验结果优化发酵培养条件,确定菌种的最佳发酵工艺,以收获最多的K88菌毛蛋白。研究表明,K88大肠杆菌在玉米浆培养基C／N、K2HP04／KH2PO4的配比分别为5／11、1／1,Mg^2＋为0．1g/mL,pH值为7．2,转速为200r／min,接种量为4．5％的条件下发酵26个小时,菌体和菌毛生产量均达到高峰,同时得出菌毛蛋白产生量和菌体量成正相关。     

巨大芽孢杆菌（Bacillus megaterium）突变株Bn,B5的生物学特性及在Vc二步发酵中的伴生作用

对选出的巨大芽孢杆菌突变株Bn,B5进行了生物学特性及发酵条件的研究,发现它们具耐低pH和抗高浓2KGA特性.可促进氧化葡萄糖酸杆菌生长,使其延迟期缩短,产酸增加.适宜的通气量下,摇瓶糖酸转化率提高10%～14%;当发酵pH为6.2～6.6时,转化率提高20%～30%.     

灵芝液体发酵过程中菌体形态与胞外多糖产量的关系

灵芝菌体形态在发酵过程存在由丝状体（０－４８ｈ）到球状体（４８ｈ以后）,再到部分丝状体的转变（７２ｈ开始）。以葡萄糖,玉米粉、酒糟、酵母膏和麸皮为碳源和氮源时的实验结果表明：菌球直径小,数量多,松散和丝状体比例高,有利于胞外多糖的形成。较高的摇瓶转变速和接种量有利于多糖的形式。     

Sperm competition mechanisms in birds: models and data

This study presents three models to explain the mechanism oflast male sperm precedence in birds. Because passive loss ofsperm from the female reproductive tract occurs, all modelsincorporate this process. The three models are passive spermloss alone, stratification with passive sperm loss, and displacementwith passive sperm loss. With two inseminations containing thesame number of sperm, the models make the following predictions.For passive sperm loss alone, (1) differential paternity ispositively and linearly related to the time interval betweeninseminations, (2) with a slope that is equal to rate of lossof sperm from the female reproductive tract, (3) with an interceptthat is the same as the differential fertilizing capacity betweenthe semen of the two inseminations, and (4) the ratio of offspringfrom two inseminations remains constant over time. For stratification,(1) the relationship between differential paternity and theinterval between inseminations is nonlinear and exhibits a "brokenstick" pattern, with a substantial first-insemination precedencefor short intervals, and (2) the proportion of offspring fatheredby the first insemination increases over time. For displacement,the relationship between differential paternity and the intervalbetween inseminations is nonlinear and also exhibits a "brokenstick" pattern, but in contrast to the stratification model,sperm from the last insemination have precedence. Data fromthree experimental studies of the domestic fowl and one forthe turkey provide the opportunity to test these models, albeitto different extents. The data from all studies are consistentwith the passive sperm-loss model, except that one aspect ofone data set provided ambiguous support for stratification.None of the data provided any support for the displacement model.     

Mechanisms Prevalent during Bioremediation of Wastewaters from the Pulp and Paper Industry

Bioremediation of wastewaters represents an important treatment methodology, especially when examined against the backdrop of ever-stricter legislation that is evolving in order to regulate effluent release into the environment. It has been reported that bioremediation specifically holds promise in solving environmental problems. Crucial questions surrounding the treatment of effluents include: efficiency of the process, economic feasibility, legal requirements, and the mechanisms involved in the remediation process. Of all these issues mentioned, the last requires special attention. This paper investigates these matters and focuses on techniques that are currently employed to determine the efficiency of bioremediation and mechanisms involved therein. The physiological significance of biosorption is also examined, as this subject has not been fully addressed in previous publications.     

沙地云杉小树高生长与环境条件关系的数量化模型

沙地云杉小树高生长与环境条件关系的数量化模型郑元润徐文铎（中国科学院植物研究所,北京１０００９３）（中国科学院沈阳应用生态研究所,１１００１５）ＴｈｅＭｏｄｅｌｓｏｆＲｅｌａｔｉｏｎｓｈｉｐｂｅｔｗｅｅｎＨｅｉｇｈｔＧｒｏｗｔｈｏｆＰｉｃｅａｍｏｎｇ...