无机离子和有机溶质对α-淀粉酶热稳定性的影响

长期以来,如何提高酶蛋白的热稳定性是分子生物学、生物工程学、化学工业等所关注的重要研究课题之一。分析了多种无机离子、糖和氨基酸对枯草杆菌液化型α-淀粉酶热稳定性的影响以及它们的共存效应,获取了一些对相关研究领域具有理论参考和实际应用价值的实验结果。在无机盐中,1mmol/L的钙离子或50mmol/L的钠离子能显著地提高该酶的热稳定性;酸性氨基酸和碱性氨基酸表现出相反的结果：酸性氨基酸具有明显的增强作用,碱性氨基酸却使之降低;随着糖浓度的增加（0～1000mmol/L）,该淀粉酶的热稳定性呈线性增高;当钠离子或钾离子与某些氨基酸或糖类共同存在时,对该淀粉酶的热稳定性表现出了明显的协同作用。试图通过检测酶蛋白分子荧光强度改变来反映该酶的热稳定性变化,其结果是：随着温度的改变,酶蛋白的荧光强度的衰减与残余酶活性之间显示了良好的相关性。从而说明热环境使酶蛋白分子的螺旋结构发生变化而失活,某些溶质的存在可能是通过作用于蛋白质分子的立体结构而影响该酶的热稳定性。     

含度量误差的黑龙江省主要树种生物量相容性模型

基于516株样木的生物量数据,采用非线性度量误差模型理论和方法,构建了黑龙江省15个主要树种(组)总生物量与地上、地下、树干、树冠、树枝、树叶6个分项生物量以及分项生物量间的相容性生物量模型,分别选出各树种总生物量和各分项生物量的最优模型,采用比值函数分级联合控制方程组构建了以总生物量为基础的相容性模型,并采用对数变换对总生物量模型消除异方差,采用加权回归对各分项生物量模型消除异方差.结果表明:本文所建的15个树种(组)相容性生物量模型中,总生物量的预估精度最高,达到90％以上;其次是地上部分生物量和树干生物量,预估精度在87.5％以上;地下部分、树冠、树枝和树叶生物量的预估精度相对较低,但绝大多数树种(组)的预估精度在80％以上;所有树种(组)总生物量、地上部分生物量、树干生物量模型的模拟效率(EF)值达0.9以上,绝大多数树种(组)的地下部分、树冠、树枝、树叶生物量模型的EF值在0.8以上.     

Differential regulation in tobacco cell suspensions of genes involved in plant-bacteria interactions by pathogen-related signals

Six cDNA clones whose corresponding mRNAs accumulate early during the hypersensitive reaction in tobacco leaves have been classified into 2 groups according to their maximum levels of accumulation in an incompatible versus a compatible interaction withPseudomonas solanacearum. We present evidence that, at least in the first stages of the interaction, tobacco cell suspensions retain the ability to respond differentially to compatible and incompatible isolates ofP. solanacearum.In addition, studies on the effect of a fungal elicitor on the accumulation of the mRNAs corresponding to the cDNA clones in cell suspensions indicate that only one group of genes responds to this treatment.     

Osmotic relations of the drought-tolerant shrub Artemisia tridentata in response to water stress

Turgor maintenance, solute content and recovery from water stress were examined in the drought-tolerant shrub Artemisia tridentata. Predawn water potentials of shrubs receiving supplemental water remained above ?2 MPa throughout summer, while predawn water potentials of untreated shrubs decreased to ?5 MPa. Osmotic potentials decreased in conjunction with water potentials maintaining turgor pressures above 0 MPa. The decreases in osmotic potentials were not the result of osmotic adjustment (i.e. solute accumulation). Leaf solute contents decreased during drought, but leaf water volumes decreased more than 75% from spring to summer, thereby passively concentrating solutes within the leaves. The maintenance of positive turgor pressures despite decreases in leaf water volumes is consistent with other studies of species with elastic cell walls. Inorganic ion, organic acid, and carbohydrate contents of leaves declined during drought. The only solutes accumulating in leaves of A. tridentata with water stress were proline and a cyclitol, both considered compatible solutes. Total and osmotic potentials recovered rapidly following rewatering of shrubs; solute contents did not change except for a decrease in proline. Maintaining turgor through the passive concentration of solutes may be advantageous compared to synthesis of new solutes for osmotic adjustment in arid environments.     

Osmolytes modulate polyglutamine aggregation in a sequence dependent manner

Osmolytes stabilize protein structure and suppress protein aggregation. The mechanism of how osmolytes impact polyglutamine (polyQ) aggregation implicated in Huntington's disease was studied. By using a reverse‐phase chromatography assay, we show that methylamines‐trimethylamine N‐oxide and betaine are generic in enhancing polyQ aggregation, while a disaccharide trehalose and an amino acid citrulline moderately retard polyQ aggregation in a sequence specific manner. Despite the altered kinetics, the fundamental nucleation mechanism of polyQ aggregation and the nature of end stage aggregates remains unaffected. These results highlight the importance of using osmolytes as modulatory agents of polyQ aggregation.     

Osmoregulatory responses of bacteria isolated from fresh or composted,olive-mill waste-waters

Halotolerant bacteria isolated from raw, olive-mill waste-waters (alpechin) and from composted alpechin could grow on solid medium containing up to 10% (w/v) NaCl. Most (70%) of these halotolerant isolates could also grow in liquid minimal medium with the same NaCl concentration and three isolates from this group were chosen for further study. When grown in tryptic soy broth (TSB), two isolates responded to lowering of water activity (a _w) by addition of NaCl or sucrose in the expected manner: by increasing the proportion of membrane anionic lipids diphosphatidylglycerol or phosphatidylglycerol. Some solute-specific differences were observed. In contrast, the third isolate did not alter its membrane phospholipid composition significantly in response to growth in NaCl, whereas in sucrose there was an increase in phosphatidylethanolamine. This response is contrary to the accepted interpretation of the function of such a _w-dependent changes, as being a mechanism for preserving the membrane lipid-bilayer phase. When all three isolates were grown in the presence of alpechin, there was a decrease in the proportion of phosphatidylglycerol and a rise in the level of phosphatidylethanolamine. Quantitative and qualitative differences in compatible solute composition were observed when the three isolates were grown in TSB with NaCl or sucrose added to lower a _w. The major compatible solutes in two of the isolates were proline and betaine, whereas in the third they were proline, betaine and ectoine; one isolate also contained some trehalose when NaCl but not sucrose was the osmolyte.     

Proline and glycine betaine accumulation in two succulent halophytes under natural and experimental conditions

Proline (Pro) and glycine betaine (GB) contents were determined in two Mediterranean halophytes, Plantago crassifolia and Inula crithmoides, to assess their possible role in salt tolerance of both taxa. Plant material was collected in a littoral salt marsh under different environmental conditions, and from plants subjected to salt treatments in a growth chamber. Relative growth inhibition by NaCl indicated that I. crithmoides is more salt-tolerant than P. crassifolia, in agreement with the distribution of the two species in nature. Field and laboratory data confirmed GB as the major osmolyte responsible for osmotic adjustment in I. crithmoides, but with only a minor role – if any – as “osmoprotectant” in the salt tolerance of P. crassifolia. Under natural conditions, Pro contents were very low in both taxa, but increased to levels high enough to contribute significantly to osmotic balance when plants were artificially treated with 450–600 mM NaCl – higher salt concentrations than those they would normally encounter in their natural habitats. These data suggest that halophytes possess built-in mechanisms, such as accumulation of additional osmolytes, to rapidly adapt to increasing salinity levels in their natural ecosystems; for example, those expected to be caused by climate change in salt marshes in the Mediterranean region.     

Compatible solutes and their effects on phosphoenolpyruvate carboxylase of C4-halophytes

Abstract Phosphoenolpyruvate carboxylase (PEPCase), extracted from two Poaceae (Cynodon dactylon and Sporobolus pungens) grown on saline soil, was affected physiologically by betaine and proline. Its affinity for phosphoenolpyruvate (PEP) was increased and full protection against NaCl inhibition was observed; enzymic activity was also stabilized when assayed at low PEP levels. Betaine has the same effects on PEPCase extracted from a Chenopodiaceae (Salsola soda), whereas proline behaves as a competitive inhibitor, i.e. it does not protect the enzyme against NaCl and it accelerates inactivation at low PEP levels. Betaine was only compatible with PEPCase extracted from Saisola kali, without any effect on activity, protection or stabilization, but proline was again inhibitory. The levels of free proline in the two salt-stressed Poaceae were high, whereas in the Chenopodiaceae the free proline was low, as in non-stressed plants. The above data indicate that osmoregulators could not only be compatible with cytoplasmic enzymes, but they could either promote or inhibit enzyme activity, depending on the source of enzyme. Coevolution of PEPCase with the osmolyte selected for, could also be inferred.     

Definition of physical integrity of synaptosomes and myelosomes by enzyme osmometry

Isotonic requirements for synaptosomes were shown to vary with the concentration of sucrose or mannitol in the isolation medium, as well as with their differential permeability to polyols and ions. The technique of enzyme osmometry, which permits quantitation of the osmotic integrity in a heterogeneous population, was used to defined the osmotic requirements for synaptosomes and myelosomes in a variety of ionic and non-electrolyte media. Important differences, observed in the rank order of permeability of synaptosomal and myelosomal membranes to electrolyte media, were consistent with the known channel density/electrical activity of the corresponding plasma membranes.     

The compatibility of osmotica in cyanobacteria

Abstract. The solutes accumulated by cyanobacteria in response to hyper-osmotic stress include Na⁺, K⁺, sucrose, trehalose, glucosyl-glycerol, glyeine betaine and glutamate betaine. The compatibility of several of these solutes with glutamine synthetase activity has been examined using cell-free extracts from a range of freshwater, marine and halotolerant cyanobacteria. All of the solutes tested were compatible with (i.e. non-inhibitory to) enzymic activity at physiological concentrations and the results demonstrate a rank order of compatibility which correlates with the concentrations at which the organic solutes occur in cyanobacteria, i.e. glycine betaine > polyol-derivatives > disaccharides and with the upper salinity limit for growth. The protection against inhibition by NaCl (halo-protection) afforded by these solutes to enzymic activity was also examined. Only glycine betaine was found to exert a significant halo-protective effect and this may be explained by differences in the mechanism of compatible solute function between small charged molecules and sugars/polyols.