Properties of enzymes from bacteria grown in the 70–100°C range

In chemostat cultures of Bacillus caldolyticus, adaptation in a single step from 70–100°C was followed under aerobic and oxygen-limited conditions and was found to proceed more smoothly under the latter circumstances. Variations of the medium (e.g. yeast extract or silicate concentrations) showed that growth at 100°C is in all respects similar to that of cultures at moderate temperatures.Enzyme preparations derived from cultures at 5°C intervals between 70 and 100°C were used to determine the temperature range. For all nine enzymes tested, the optimum temperature was found to be 67°C; the latter was independent of the growth temperature. Differences were found, however, with respect to the maximum temperature of individual enzymes, and three groups, with maxima between 70 and 80°C, 80 and 90°C and 90 and 100°C can be distinguished. Again, there was no correlation with the growth temperature.Stability experiments also revealed that enzymes from the same organism can have different thermal properties: Some were found to be quite thermolabile (e.g. the pyruvate kinase), while others (e.g. hexokinase and glutamate-pyruvate transaminase) exhibited a high thermostability. These properties were not related to the growth temperature within the 70–100°C range, too.Six of the enzymes tested could be stabilized by their respective substrates, but the degree of protection varied for individual enzymes. Three enzymes (acetate kinase, glutamate dehydrogenase and myokinase) could not be stabilized by their substrates.Comparative experiments with the hexokinase suggested, that the thermal integrity of the enzymes is better protected within the cell as compared to the stability of the enzyme preparations.Abbreviations used AK acetate kinase - Ala-DH alanine dehydrogenase - Ald aldolase - GIDH glutamate dehydrogenase - G6P-DH glucose-6-phosphate dehydrogenase - GTP glutamate-pyruvate transaminase - HK hexokinase - ICDH isocitrate dehydrogenase - MK myokinase - PK pyruvate kinase     

乙型脑炎病毒NS2A-C60A位点突变对其生物学特性影响研究*

目的:旨在探索Ⅰ型日本乙型脑炎病毒传代致弱后基因组突变NS2A-C60A对乙脑病毒生物学特性的影响。方法:首先通过对传代致弱及原始乙脑毒株基因组序列进行测序比对、结构预测分析并利用Western blotting(WB)确定了目标研究位点NS2A-C60A;然后使用反向遗传定点突变技术构建拯救了包含NS2A-C60A单点突变的病毒株;最后利用噬斑形态观察、生长曲线、双萤光素酶分析,WB以及炎性因子检测和动物实验研究了该单点突变对于乙脑病毒生物学特性的影响。结果:首次研究发现Ⅰ型乙脑病毒传代致弱会导致NS1'蛋白表达的显著下降以及可能的相关位点NS2A-C60A,并成功拯救获得了NS2A-C60A单点突变毒株rJEV-C60A,研究发现NS2A-C60A突变对乙脑病毒的生长特性及噬斑形成没有显著影响,但是能够显著降低乙脑病毒NS1'蛋白的表达,并且该位点突变能够轻微阻碍乙脑病毒对细胞炎性因子表达的抑制,动物实验结果显示NS2A-C60A点突变病毒与原毒株具有相似的神经毒力,说明该位点突变不是影响乙脑病毒毒力致弱的关键位点。结论:新发现的NS2A-C60A位点突变能够显著减少乙脑病毒NS1'蛋白的表达,但是对其增殖、诱导炎症及神经毒力等生物学特性没有显著影响。     

A proposed model for rhodopsin in photoreceptor membranes

Transient electric birefringence studies have been made on bovine rhodopsin solubilized in the detergent lauryldimethylamine oxide from glutaraldehyde fixed rod outer segment (ROS) membranes. It was found that fixation caused no appreciable differences in the measured relaxation times when compared with unfixed ROS. On the basis of these findings a model for the orientation of rhodopsin in photoreceptor membranes is proposed which accounts for translational diffusion and two modes of rotational diffusion. The proposed model is related to a number of experimentally determined biophysical properties reported in the literature.     

Spatial patterns in coupled biochemical oscillators

Summary The effects of diffusion on the dynamics of biochemical oscillators are investigated for general kinetic mechanisms and for a simplified model of glycolysis. When diffusion is sufficiently rapid a population of oscillators relaxes to a globally-synchronized oscillation, but when diffusion of one or more species is slow enough, the synchronized oscillation can be unstable and a nonuniform steady state or an asynchronous oscillation can arise. The significance of these results vis-a-vis models of contact inhibition and zonation patterns is discussed.     

木犀科植物叶绿体基因组结构特征和系统发育关系

木犀科11属19个种叶绿体基因组的一般特征和变异特征的比较分析显示, 结果表明, 该科叶绿体基因组大小为154-165 kb, 其差异主要是大单拷贝(LSC)长度的差异所致。Jasminum属3个物种的叶绿体基因组长度与其余物种有较大差异, 该属clpP基因内含子和accD基因丢失。共线性分析表明, Jasminum属3个物种多个基因出现基因重排现象, 倒位可能是重排的主要原因。Jasminum属在IRb/SSC和SSC/IRa边界的基因均与其它物种不同; 重复序列与SSR数量检测结果表明, Jasminum属与其余物种在数量及重复长度上差异较大。基于CDS数据构建的系统发育树表明, Abeliophyllum distichum和Forsythia suspensa为木犀科中较早分化的类群。     

黄腐酸增强小麦抗旱能力的生理生化机制初探

叶面喷施黄腐酸可显著提高小麦幼苗的保水能力,表现为降低叶面蒸腾强度,增加气孔扩散阻力,提高幼苗的生物量,在干旱条件下尤为明显。喷施黄腐酸可使干旱条件下叶片内脯氨酸含量提高近一倍,并在水分充足时,也能使叶片脯氨酸含量增加78%。     

新型隐球菌L型生物学性状的研究

本文制备了新型隐球菌L型高渗培养基。新型隐球菌L型的特征为:革兰氏染色性改变;大小不一;能通过孔径0.8μm的滤器;电镜检查细胞壁有不同程度的缺损。在高渗液体培养基中,新型隐球菌L型缓慢生长、不易回复;在高渗平板上则易回复。稳定、不稳定隐球菌L型的生化反映与其酵母型相似,但稳定L型的同化能力及尿素分解能力减弱。新型隐球菌形成L型后,其形态、培养特性及生化反映发生变异,这在菌种鉴定与临床检验时要注意。     

Spoil characteristics and vegetation development of an age series of mine spoils in a dry tropical environment

A series of coal mine spoils (5, 10, 12, 16 and 20-yr old) in a dry tropical environment was sampled to assess the changes with time in spoil characteristics, species composition and plant biomass. Coarse fragments (>2 mm) decreased with age of mine spoil while the proportion of 0.2–0.1 mm particles increased. Total soil N, mineral N, NaHCO₃-extractable Pi, and exchangeable K increased with age of mine spoil and these parameters were lower in mine spoils than native forest soil even after 20 years of succession. Exchangeable Na decreased with age of mine spoil and in 20-yr old spoil it was higher than native forest soil. Plant community composition changed with age. Only a few species participated in community formation. Species richness increased with age, while evenness and species diversity declined from 5-yr old to 16-yr old community with an increase in the 20-yr old community. A reverse trend occurred for concentration of dominance. Area-weighted shoot and root biomass of other species increased with the age of the mine spoil while that of Xanthium strumarium patches declined with age. Data collected on spoil features, microbial C, N and P, and shoot and root biomass when subjected to Discriminant Analysis indicated a continued profound effect of age. 10 and 12-yr old mine spoils were closer to each other, and 5 and 20-yr old spoils were farthest apart.     

The acquisition of inorganic carbon by four red macroalgae

Photosynthesis was studied in four species of red marine macroalgae: Palmaria palmata, Laurencia pinnatifida, Lomentaria articulata and Delesseria sanguinea. The rate of O₂ evolution for submersed photosynthesis was measured as a function of incident photon flux density at normal pH and inorganic carbon concentration (pH 8.0, 2 mol m^–3), and as a function of inorganic carbon concentration at pH 8.0 at saturating and at limiting photon flux density. The rate of CO₂ uptake was measured for emersed photosynthesis as a function of CO₂ partial pressure at saturating photon flux density. Previous pH-drift results suggest that Palmaria and Laurencia are able to use HCO _inf3 ^sup– as well as CO₂ whereas Lomentaria and Delesseria are restricted to CO₂. None of the algae are saturated by 2 mol m^–3 inorganic carbon at high light (400 mol m^–2 s^–1) but are saturated at low light (35 mol m^–2 s^–1). The inorganic C concentration at which half the light-saturated rate of O₂ evolution is achieved is higher for Palmaria and Laurencia (1.51 and 1.85 mol m^–3) than for Lomentaria and Delesseria (0.772 and 0.841 mol m^–3). The lower values for the latter two species could reflect their putative restriction to CO₂. If expressed in terms of CO₂, the half-saturation values yield 7.2 and 7.8 mmol m^–3 respectively, which are very similar to values obtained previously during pH-drift experiments but at lower concentrations of HCO _inf3 ^sup– , consistent with restriction to CO₂. The photosynthetic conductance (m s^–1), calculated from the initial slope for photosynthesis at low concentrations of inorganic carbon, correlates with the suggested ability to extract inorganic carbon based on pH-drift results. Calculations made assuming that CO₂ is the only species diffusing across the boundary layer are consistent with boundary layer thicknesses of 20 and 19 m for Lomentaria and Delesseria respectively, which is feasible given the rapid water movement in the experiments. For Laurencia however, an unreasonably small boundary layer thickness of 6 m is necessary to explain the flux, which indicates co-diffusion by HCO _inf3 ^sup– . In the apparent absence of external carbonic anhydrase, direct uptake of HCO _inf3 ^sup– , rather than external conversion to CO₂ is indicated in this species. In air, the CO₂ concentration at which photosynthesis is half-maximal increases in the same order as the ability to raise pH in drift experiments. At 21 kPa the CO₂ compensation partial pressures for Palmaria and Laurencia at 0.56 and 1.3 Pa are low enough to suggest a carbon-concentrating mechanism is operating, while those of Lomentaria at 1.8 Pa and particularly that of Delesseria at 4.5 Pa could be explained without a carbon-concentrating mechanism. The algae tested (all except Delesseria) showed more O₂ evolution than could be accounted for with a photosynthetic quotient of 1.0 and uncatalysed conversion of HCO _inf3 ^sup– to CO₂ outside the cell in high light at pH 8.0 when high algal fresh weight per unit medium was used. These results are concordant with other data suggesting use of HCO _inf3 ^sup– by Palmaria and Laurencia, but discordant with the rest of the available information in indicating use of HCO _inf3 ^sup– by Lomentaria. The reason for this is unclear. The lightsaturated rate of O₂ evolution on an algal area basis and the photon flux density needed to saturate photosynthesis were related partly to the habitat from which the seaweeds were collected, but more strongly to the ability to use HCO _inf3 ^sup– . Values for the two users of HCO _inf3 ^sup– , Palmaria (population used was intertidal; also occurs subtidally) and Laurencia (intertidal/shaded intertidal), were greater than for Lomentaria (shaded intertidal), which was greater than Delesseria (subtidal), both of which are believed to be restricted to CO₂. In accordance with earlier ¹³C data and, for Delesseria, estimates of the achieved growth rates in situ, carbon is likely to be saturating and use of HCO _inf3 ^sup– is unlikely to occur in the normal low-light habitats of Lomentaria and Delesseria. Analysis of N-use efficiencies show that they are closer to the low-CO₂-affinity Laminariales than the high-CO₂-affinity Fucaceae.