离子注入选育高产木聚糖酶黑曲霉及其发酵条件研究

以黑曲霉A3为出发菌,利用离子注入技术选育出一株遗传性状稳定的木聚糖酶高产突变株AN497,其产酶水平较出发菌从野生型A3菌株的405.6IU/ml提高到586.2IU/ml,即酶产量增加了44.5%;对高产菌进行发酵条件优化,发现以玉米芯粉为主要碳源、用蔗糖代替葡萄糖作为附加碳源,对木聚糖酶的发酵具有明显的促进作用;采用复合的无机氮源 (NH4)2SO4和NaNO3,(1: 2)浓度以10g/L为宜;菌株对发酵通氧量具有较高的要求,摇瓶转速在230r/min时的产酶水平较200r/min要高;通过发酵条件的优化,高产菌株的产酶活力最高可达671.1IU/mL,比出发菌株的产酶量提高了65.5%。     

盐胁迫下不同基因型番茄可溶性物质含量的变化

以两种野生番茄秘鲁番茄(Lycopersicon peruvianum LAlll)、潘那利番茄(Lycopersicon pennellii LA716)和两种栽培番茄"鲜丰98-7"、"交农1号"为材料,用NaCl 150 mmol/L进行盐胁迫,测定及分析了植株体内Na 、K 、Ca2 等无机离子和某些有机溶质游离脯氨酸、可溶性糖的含量.结果表明,盐胁迫条件下,野生番茄在新叶中积累了大量的Na 等无机离子,而栽培番茄新叶内脯氨酸和可溶性糖的含量却显著提高,且它具有将吸收的Na 区域化到老叶中的能力.说明野生番茄和栽培番茄在可溶性物质的合成和积累能力上有明显差异,它们是通过不同代谢途径和机理来适应盐环境的.     

Using lanthanide ions to align troponin complexes in solution: order of lanthanide occupancy in cardiac troponin C

The potential for using paramagnetic lanthanide ions to partially align troponin C in solution as a tool for the structure determination of bound troponin I peptides has been investigated. A prerequisite for these studies is an understanding of the order of lanthanide ion occupancy in the metal binding sites of the protein. Two-dimensional [(1)H, (15)N] HSQC NMR spectroscopy has been used to examine the binding order of Ce(3+), Tb(3+), and Yb(3+) to both apo- and holo-forms of human cardiac troponin C (cTnC) and of Ce(3+) to holo-chicken skeletal troponin C (sTnC). The disappearance of cross-peak resonances in the HSQC spectrum was used to determine the order of occupation of the binding sites in both cTnC and sTnC by each lanthanide. For the lanthanides tested, the binding order follows that of the net charge of the binding site residues from most to least negative; the N-domain calcium binding sites are the first to be filled followed by the C-domain sites. Given this binding order for lanthanide ions, it was demonstrated that it is possible to create a cTnC species with one lanthanide in the N-domain site and two Ca(2+) ions in the C-domain binding sites. By using the species cTnC.Yb(3+).2 Ca(2+) it was possible to confer partial alignment on a bound human cardiac troponin I (cTnI) peptide. Residual dipolar couplings (RDCs) were measured for the resonances in the bound (15)N-labeled cTnI(129-148) by using two-dimensional [(1)H, (15)N] inphase antiphase (IPAP) NMR spectroscopy.     

燃烧植物产生烟与热的生理生态效应

燃烧植物产生的烟与热对植物的生理生态功能有重要的影响,相关研究已成为生态学研究的热点之一。植物源烟对一些植物种子的萌发和幼苗生长有促进作用,这种促进作用与GA和细胞分裂素的作用相似。在植物烟水溶液中分离得到了具有促进植物种子萌发作用的化合物丁烯羟酸内酯,该物质具有热稳定性、挥发性和有效浓度范围广等特点。丁烯羟酸内酯可以通过纤维素加热产生,因而几乎所有的植物燃烧产生的烟中都可以产生丁烯羟酸内酯。热因子对植物种子萌发有利作用表现为打破种子休眠、清除限制种子萌发的物理、化学因素和减轻种子病原体等方面。大量研究显示,不同植物对烟与热的响应机理存在显著的差异,这是植物群落过火后物种组成改变的重要原因之一。烟与热因子对植物生理生态作用的研究我国开展较少．这与我国是一个森林、草原火灾频繁的国家是不相称的,加强这方面的研究很有必要。另外,今后我国可以在烟与热因子对植物作用的机理,揭示传统用烟火处理土壤促进农林业植物生长的物理和化学本质,以及这些机理在发展有机农业中运用等方面开展深入的研究。     

Effects of Arachidonic Acid on the Lysosomal Ion Permeability and Osmotic Stability

In this study, we investigated the effects of arachidonic acid, a PLA₂-produced lipid metabolite, on the lysosomal permeability, osmotic sensitivity and stability. Through the measurements of lysosomal β-hexosaminidase free activity, membrane potential, intralysosomal pH, and lysosomal latency loss in hypotonic sucrose medium, we established that arachidonic acid could increase the lysosomal permeability to both potassium ions and protons, and enhance the lysosomal osmotic sensitivity. As a result, the fatty-acid-promoted entry of potassium ions into the lysosomes via K⁺/H⁺ exchange, which could produce osmotic imbalance across their membranes and osmotically destabilize the lysosomes. In addition, the enhancement of lysosomal osmotic sensitivity caused the lysosomes to become more liable to destabilization in osmotic shock. The results suggest that arachidonic acid may play a role in the lysosomal destabilization.     

Biotic indirect effects: a neglected concept in invasion biology

Indirect effects involve more than two species and are defined as how one species alters the effect that another species has on a third. These complex interactions are often overlooked in studies of interactions between alien and native species, and their role in influencing biological invasions has been rarely considered. Based on a comprehensive review of the invasion biology literature, we examine the evidence for the occurrence of four of the most commonly documented indirect effects (apparent competition, indirect mutualism/commensalism, exploitative competition, and trophic cascades) in the invasion process. Studies investigating indirect effects in the context of invasion biology are relatively rare, but have been increasing in recent years, and there are sufficient examples to indicate that this kind of interaction is likely to be more common than is currently recognized. Whether indirect interactions are mediated by an alien or a native species, and whether they occur between ecologically similar or dissimilar alien and native species, depends in part on the type of interaction considered and no predictable patterns were detected in the literature. Further empirical studies will help to elucidate such patterns. At this stage, the inherent unpredictability of indirect interactions means that their impacts in relation to invasions are particularly challenging for land managers to deal with, and their role in invasions is a complex, but is a valuable area of investigation for researchers.     

Effect of Halide Ions on t-[35S]Butylbicyclophosphorothionate Binding

The binding of t-[35S]butylbicyclophosphorothionate [( 35S]TBPS) to a site on the GABAA receptor complex is ion dependent. This study was conducted to determine the effects of ion species and concentration on the time course, affinity, and number of sites of [35S]TBPS binding. At a concentration of 200 mM ion, the time to equilibrium for [35S]TBPS binding was shortest for I-, followed by Br- less than Cl- less than F-. A similar rank order was observed for the concentration of ion required to produce half-maximal [35S]TBPS binding. Saturation binding experiments were conducted to evaluate the effect of increasing ion concentration on the KD and Bmax of [35S]TBPS binding. The Bmax was independent of both ion species and concentration. The receptor affinity, however, increased with increasing concentration for each ion. Calculated maximal affinity values were not different between ions; however, the EC50 to produce those values was different among ions and ranked in the same order as that for time course and maximal binding data. Association and dissociation rates for [35S]TBPS binding were greater in I- than in Cl-. These data emphasize the importance of ion selection and incubation times on [35S]TBPS binding.     

Characterization of protein glycosylation using chip-based nanoelectrospray with precursor ion scanning quadrupole linear ion trap mass spectrometry.

Glycosylation is one of the most important posttranslational modifications affecting the functions of proteins and cell activities. Mass spectrometry (MS) has proven to be an effective tool for structural glycobiology and has helped gain an understanding of glycoprotein-mediated diseases. Although electro-spray ionization-tandem MS remains widely recognized as an effective means for oligosaccharide characterization, the hydrophilic nature of glycans has often caused the poor ionization efficiency requiring either derivatization or nanoelectrospray to improve detection sensitivity. In this report we describe the use of a chip-based infusion nanoelectrospray platform coupled with the hybrid triple quadrupole/linear ion trap for identification and characterization of glycosylation in complex mixtures. The high-mannose-type N-glycosylation in ribonuclease B was used to map the glycosylation site and obtain glycan structures. Using the chip-based nanoelectro-spray with precursor ion scanning linear ion trap MS, we were able to map the glycosylation site and obtain the glycan structures in ribonuclease B at 100 fmol/microL in a single analysis. In addition, a new, low-abundant glycoform with an additional hexose (Hex10GlcNAc2) attached to ribonuclease B was discovered. The results reported here demonstrate that the chip-based infusion nanoelectrospray ionization coupled to a quadrupole/linear ion trap platform is a valuable system, as it provides high sensitivity and stability for nanoelectrospray analysis, and allows extended acquisition time for completing precursor ion scanning and subsequent MS2 and MS3 information in a single analysis.     

Synthetic and mechanistic studies on ruthenium dicarbonyl complexes containing PhP(CH2CH2CH2PCy2)2 (Cyttp) ligand

A synthetic and mechanistic study is reported on ligand substitution and other reactions of six-coordinate ruthenium(II) carbonyl complexes containing tridentate PhP(CH₂CH₂CH₂PCy₂)₂ (Cyttp). Carbonylation of cis-mer-Ru(OSO₂CF₃)₂(CO)(Cyttp) (1) affords [cis-mer-Ru(OSO₂CF₃)(CO)₂(Cyttp)]O₃SCF₃ (2(O₃SCF₃)) and, on longer reaction times, [cis-mer-Ru(solvent)(CO)₂(Cyttp)](O₃SCF₃)₂ (solvent = acetone, THF, methanol). 2(O₃SCF₃) reacts with each of NaF, LiCl, LiBr, NaI, and LiHBEt₃ to yield [cis-mer-RuX(CO)₂(Cyttp)]⁺ (X = F (3), Cl (4), Br (5), I (6), H (7)), isolated as 3-7(BPh₄). These conversions proceed with high stereospecificity to afford only a single isomer of the product that is assigned a structure in which the Ph group of Cyttp points toward the CO trans to X (anti when X = F, Cl, Br, or I; syn when X = H). Treatment of 2(O₃SCF₃) with NaOMe and CO generates the methoxycarbonyl complex [cis-mer-Ru(CO₂Me)(CO)₂(Cyttp)]⁺ (8), whereas addition of excess n-BuLi to 2(O₃SCF₃) in THF under CO affords mer-Ru(CO)₂(Cyttp) (9). The two ¹³C isotopomers [cis-mer-Ru(OSO₂CF₃)(CO)(¹³CO)(Cyttp)]O₃SCF₃ (2′(O₃SCF₃): ¹³CO trans to P_C; 2″(O₃SCF₃): ¹³CO cis to all P donors) were synthesized by appropriate adaptations of known transformations and used in mechanistic studies of reactions with each of LiHBEt₃, NaOMe/CO, and n-BuLi. Whereas LiHBEt₃ reacts with 2′(O₃SCF₃) and 2″(O₃SCF₃) to replace triflate by hydride without any scrambling of the carbonyl ligands, the corresponding reactions of NaOMe-CO are more complex. The methoxide combines with the CO cis to triflate in 2, and the resultant methoxycarbonyl ligand ends up positioned trans to the incoming CO in 8. A mechanism is proposed for this transformation. Finally, treatment of either 2′(O₃SCF₃) or 2″(O₃SCF₃) with an excess of n-BuLi leads to the formation of the same two ruthenium(0) isomers of mer-Ru(CO)(¹³CO)(Cyttp). These products represent, to our knowledge, the first example of a syn-anti pair of isomers of a five-coordinate metal complex.     

Effects of extremely low frequency electromagnetic fields on membrane-associated enzymes

The effects of extremely low frequency electromagnetic fields of 75 Hz were studied on different membrane-associated enzymes. Only the activities of three enzymes out of seven exposed to the field decreased approximately of about 54-61% with field amplitudes above a threshold of 73-151 microT depending on the enzyme. The same field had no effect on the activities of either integral membrane enzymes such as Ca,ATPase, Na/K,ATPase, and succinic dehydrogenase or peripheral membrane enzymes such as photoreceptor PDE. The decrease in enzymatic activity of the field-sensitive enzymes was independent of the time of permanence in the field and was completely reversible. When these enzymes were solubilized with Triton, no effect of the field was obtained on the enzymatic activity, suggesting the crucial role of the membrane in determining the conditions for enzyme inactivation. The role of the particular linkage of the field-sensitive enzymes to the membranes is also discussed.