Dye-ligand chromatography for the resolution and purification of restriction endonucleases

The resolution of restriction endonucleases from the same microorganism is conventionally achieved by lengthy fractionation protocols. We now report effective single-step procedures that exploit dye-ligand chromatography for the resolution and purification of restriction enzymes. After suitable initial screening, we demonstrated that resolution of two restriction activites can be achieved in one chromatographic step, and further purification can subsequently be effected using selected dye-adsorbents. Accordingly, we resolved in one step, Hpa I from Hpa II, Hind II from Hind III, and Sac I from Sac II. Furthermore, a three-step Chromatographic procedure has been developed to purify EcoRV suitable for commercial exploitation, as judged by the “overdigestion” and “cut-ligate-recut” quality control tests.     

金属置换碳酸酐酶及其生成动力学

本文报道了金属置换碳酸酐酶的活性研究。指出了离子半径的大小,配位构型的要求,热力学稳定性和配位体交换动力学是决定各种金属置换碳酸酐酶活性的重要因素。并用金属置换法测定了Ni(Ⅱ)与apo-CA形成金属酶的反应动力学速率常数。在25℃、pH7.47时,K_(fNi(Ⅱ)-CA)为14.01·mol~(-1)sec~(-1),较之于Zn(Ⅱ)-CA的速率常数小10~3倍。Cd(Ⅱ)-CA的生成动力学为一快反应,采用配位竞争法,在邻菲罗啉配位剂存在下,测得了Cd(Ⅱ)-CA生成动力学的条件速率常数在o-phen浓度为1.96和2.92×10~(-4)mol·1~(-1)时分别为137.1和114.61·mol~(-1)sec~(-1)。这一研究提示了镉对生物体的毒性作用的一个重要方面是使锌酶失活。而配位剂的存在大大降低了镉与脱辅基锌酶的反应动力学,这也是它们显示解毒功能所在。     

毛细管电泳在拮抗药物对氨基苯甲酸、对羟基苯甲酸和磺胺分析测定中的应用

本文报道了一种用毛细管区带电泳法(CZE)分离与测定对氨基苯甲酸、对羟基苯甲酸及磺胺类药物的新方法.电泳条件为:用 20mmol/L硼砂-20mmol/L H_3PO_4-20 mmol/Lβ-环糊精-4％乙醇(pH 7.0)作电泳液,L-抗坏血酸为内标,280nm为检测波长,样品由电进样方式(10kV/10s)引入毛细管(51.2 cm×50μm i.d.,有效分离长度为 38.5 cm).在24.5°C下,6 min内三者可达基线分离(电泳电压 25kV),且在一定范围内可进行定量分析,保留时间(Tr)及A_(样品)/A_(内标)的RSD值分别小于1.0％和5.0％.本法的建立为研究这三者共存于高等动物及微生物体内时的生理作用提供了一种可共选择的新方法.     

纸张上光油关键物质的成分分析

利用色相色谱／质谱联用技术，傅立叶红外及裂解－气相色谱／质谱联用技术，对印刷用上光油的关键成分进行了分析，共鉴定出７种挥发性组分，２种高分子聚合物。     

Implementation and efficiency of an automatic peak-purity-control procedure in HPLC-UV-VIS-coupling based on principal component analysis

Summary Based on a thorough knowledge of the actual system precision significance testing of the primary eigen values, resulting from principal component analysis of the two-dimensional data array of HPLC with photodiode-array detection, is a powerful means to uncover unresolved chromatographic peaks. The implementation of this chemometric technique for assuring peak homogeneity and results showing the efficiency for two-component peaks in regard to spectral characteristics, chromatographic resolution and absorbance ratio of the investigated compounds are presented.     

Relationship between the molybdenum phases and the conversion of n-butane over Mo/HZSM-5

The conversion of n-C₄H₁₀ was undertaken on MoO₃/HZSM-5 catalyst at 773–973 K and the phases of molybdenum species were detected by XRD. The XRD results show that bulk MoO₃ on HZSM-5 can be readily reduced by n-C₄H₁₀ to MoO₂ at 773 K and MoO₂ can be gradually carburized to molybdenum carbide above 813 K. The molybdenum carbide formed from the carburization of MoO₂ with n-C₄H₁₀ below 893 K is -MoC_1−x with fcc-structure, while hcp-molybdenum carbide formed above 933 K. During the evolution of MoO₃ to MoO₂ (>773 K) or the carburization of MoO₂ to molybdenum carbide (>813 K), deep oxidation, cracking and coke deposition are serious, in particular at higher reaction temperatures, these lead to the poor selectivity to aromatics. Aromatization of n-C₄H₁₀ can proceed catalytically on both Mo₂C/HZSM-5 and MoO₂/HZSM-5, the distribution of the products for the two catalysts is similar below 813 K, but the activity for Mo₂C/HZSM-5 is much higher than that for MoO₂/HZSM-5.     

The regioselectivity of the formation of 2-pyrazolylthiazoles and their precursors from the reaction of 2-hydrazinothiazoles with 4,4,4-trifluoro-1-hetaryl-1,3-butanediones

Reaction of 2-hydrazinothiazoles 1 with 1-thienyl- and 1-furyl-1,3-butanediones 2a,b in methanol in the presence of hydrochloric acid mainly leads to a mixture of pyrazoles 3 and pyrazolines 4 or pyrazoles 3 and 5 in strong acidic conditions. Isomeric hydrazones 6 and pyrazolines 4 were formed and isolated in these reactions in the absence of hydrochloric acid. It has been shown that the regioselectivity in the reaction of diketones 2 with hydrazine 1 is governed by both the concentration of acid and the nature of substituents in the 1,3-diketones 2. Cyclization of hydrazones 6 is shown to occur under milder conditions than dehydration for pyrazolines 4. The new heterocyclic compounds were prepared and fully characterized by NMR spectra and by X-ray analysis for 3c.     

Propane/propylene separation by selective olefin adsorption on Cu/SBA-15 mesoporous silica

Samples of mesoporous silica, SBA-15, were prepared under hydrothermal conditions and Cu cations were incorporated into the framework by two different impregnation techniques. The corresponding adsorption/desorption isotherms of propylene, propane, and N₂ were measured to evaluate the material's effectiveness in the separation of propane/propylene mixtures. Adsorption uptake of propylene increased and that of propane decreased in Cu containing samples as compared to the uptakes observed in undoped SBA-15 samples. It was demonstrated that the presence of Cu atoms in the adsorbent lattice led to a greater selectivity towards propylene. Furthermore, the highest level of Cu(I) were obtained in samples prepared by equilibrium impregnation, which in turn improved the olefin/paraffin uptake ratio. Under some working conditions, the amount of propylene adsorbed in selected samples is totally reversible while propane uptake was negligible.     

Optimization of inoculum to substrate ratio for enhanced methane yield from leather fleshings in a batch study

One of the most significant issues of the last few decades has been tracing for renewable energy sources. Animal fleshing (ANFL) is the most common proteinaceous solid waste accured during the production of leather and it must be disposed of in an environmentally responsible manner. This paper is attempts to assess the biogas production from solid waste originating from the chrome based tannery. Anaerobic digestion of these wastes will be a viable option for waste stabilization and energy production in the form of biogas to be utilized in the industry. The bio-methane potential of the wastes were examined by mixing these wastes with various sources of inoculum and different inoculum to substrate (I/S) ratio considered. The batch experiments were carried out in 2.5 l glass reactors with a various source of inoculumviz., Cow Dung (CD), Elephant Dung (ED) and Bio-Digested Slurry (BDS) with varied inoculum to substrate (LFs) ratios for a retention time of 50 days with replications. The results obtained from the experiments showed that BDS:LF (25:75) had the highest gas production of 14505 ml (651.85 ml CH₄g^?1 VS) followed by CD:LF (50:50) produced 12072.5 ml (789.36 ml CH₄g^?1 VS) and ED:LF (75:25) produced 11252.5 ml (1492.08 ml CH₄g^?1 VS)with a methane content of 63.77, 61.92 and 62.72%, respectively.     

Can the presence of chemically inert species modify the face selectivity in asymmetric induction by cyclodextrins?

Reduction of-cyclodextrin (-CD) aromatic ketone (acetophenone and acetonaphthones) inclusion compounds were carried out in the presence of a large number of chemically inert species as potential co-guests. In several cases, it was observed that stoichiometric molar ratios of these compounds to ketone significantly modify the chiral induction yielding the inverted alcohol enantiomer and increasing the face selectivity. The results were found to depend strongly on the respective structure and shape of both the ketone and the additive, and on the molar ratio of-CD:ketone:third compound. These observations suggest the formation of a three-component inclusion complex in which the geometry of binding of the substrate and its mobility are changed with respect to the binary system.