粗纤维素酶液对微晶纤维素酶解的影响因素

栗生灰黑孔菌多被用于产漆酶的研究,很少有利用其纤维素酶的报道。为了降低在纤维素水解中纤维素酶的使用成本,利用栗生灰黑孔菌发酵制备的粗纤维素酶液,以微晶纤维素为底物模型,研究粗纤维素酶液水解微晶纤维素的最佳pH、温度和最佳表面活性剂助剂种类及浓度,并对不同表面活性剂存在条件下的纤维素酶解动力学、紫外和荧光光谱进行了研究。结果表明,粗纤维素酶水解微晶纤维素的最佳条件为pH 4.8,温度50℃,最佳表面活性剂助剂为吐温80,添加剂量为1.12mg/g底物;吐温80的添加可提高粗纤维素酶解的最大反应速度常数V_max,降低米氏常数K_m;表面活性剂改变了纤维素酶的紫外和荧光最大吸收峰,酰胺Ⅰ带和酰胺Ⅲ带的谱峰,可能通过与纤维素酶中的氨基酸残基发生反应影响了纤维素酶的结构,进而影响了微晶纤维素的水解反应。该研究为进一步降低纤维素水解成本提供了理论指导。     

Metal-catalyzed nitrogen-atom transfer methods for the oxidation of aliphatic C-H bonds

For more than a century, chemists have endeavored to discover and develop reaction processes that enable the selective oxidation of hydrocarbons. In the 1970s, Abramovitch and Yamada described the synthesis and electrophilic reactivity of sulfonyliminoiodinanes (RSO(2)N═IPh), demonstrating the utility of this new class of reagents to function as nitrene equivalents. Subsequent investigations by Breslow, Mansuy, and Müller would show such oxidants to be competent for alkene and saturated hydrocarbon functionalization when combined with transition metal salts or metal complexes, namely those of Mn, Fe, and Rh. Here, we trace our own studies to develop N-atom transfer technologies for C-H and π-bond oxidation. This Account discusses advances in both intra- and intermolecular amination processes mediated by dirhodium and diruthenium complexes, as well as the mechanistic foundations of catalyst reactivity and arrest. Explicit reference is given to questions that remain unanswered and to problem areas that are rich for discovery. A fundamental advance in amination technology has been the recognition that iminoiodinane oxidants can be generated in situ in the presence of a metal catalyst that elicits subsequent N-atom transfer. Under these conditions, both dirhodium and diruthenium lantern complexes function as competent catalysts for C-H bond oxidation with a range of nitrogen sources (e.g., carbamates, sulfamates, sulfamides, etc.), many of which will not form isolable iminoiodinane equivalents. Practical synthetic methods and applications thereof have evolved in parallel with inquiries into the operative reaction mechanism(s). For the intramolecular dirhodium-catalyzed process, the body of experimental and computational data is consistent with a concerted asynchronous C-H insertion pathway, analogous to the consensus mechanism for Rh-carbene transfer. Other studies reveal that the bridging tetracarboxylate ligand groups, which shroud the dirhodium core, are labile to exchange under standard reaction conditions. This information has led to the generation of chelating dicarboxylate dinuclear rhodium complexes, exemplified by Rh(2)(esp)(2). The performance of this catalyst system is unmatched by other dirhodium complexes in both intra- and intermolecular C-H amination reactions. Tetra-bridged, mixed-valent diruthenium complexes function as effective promoters of sulfamate ester oxidative cyclization. These catalysts can be crafted with ligand sets other than carboxylates and are more resistant to oxidation than their dirhodium counterparts. A range of experimental and computational mechanistic data amassed with the tetra-2-oxypyridinate diruthenium chloride complex, [Ru(2)(hp)(4)Cl], has established the insertion event as a stepwise pathway involving a discrete radical intermediate. These data contrast dirhodium-catalyzed C-H amination and offer a cogent model for understanding the divergent chemoselectivity trends observed between the two catalyst types. This work constitutes an important step toward the ultimate goal of achieving predictable, reagent-level control over product selectivity.     

MoO2 Diffusion in Aluminosilicate Glass

A diffusion couple of an oxidized molybdenum disk and a glass cylinder was used to measure the solubiiity and effective binary diffusion coefficient of MoO₂ in a non-alkali aluminosilicate glass. At 1400°C, the solubility limit was 8.4 mol%; the value of the diffusion coefficient (4.1 × 10⁻¹⁶ m²/s) was significantly lower than that estimated from the Stokes-Einstein relation.     

Ditopic, flexible hydrazone-based building blocks with pendant 2,2′:6′,2′′-terpyridine metal-binding domains

The synthesis and characterization of three new bis(2,2′:6′,2′-terpyridine) (tpy) ligands containing different hydrazone spacers between the metal-binding domains are described. Treatment of 1,4-benzenedicarbaldehyde bis(2,2′:6′,2′′-terpyridin-4′-ylhydrazone) (1) with [(tpy)RuCl₃] in the presence of N-ethylmorpholine results in the formation of [(tpy)Ru(μ-1)Ru(tpy)]⁴⁺. Single crystal X-ray diffraction data for [(tpy)Ru(μ-1)Ru(tpy)][PF₆]₄·8MeCN confirm the ability of the hydrazone-based ligand to bridge two ruthenium(II) centres, providing proof-of-principle for the application of this class of flexible ligand in the design of coordination polymers.     

Supramolecular and nanochemistry

The following contributions describe various research activities of the Department of Chemistry, University of Basel in the area of nanochemistry and supramolecular chemistry.     

Starch-zein blends formed by shear flow

A newly in-house developed shearing device was used to explore the formation of new types of microstructures in concentrated starch-zein blends. The device allowed processing of the biopolymer blends under homogeneous, simple shear flow conditions. Water and glycerol were added as plasticizers. Different ratios (0-20% zein, dry basis) were used to study the influence of the matrix composition and processing conditions on the properties of the final material. The properties at large deformation were examined by tensile tests in two different directions (i.e. along the flow (∥) and in the vorticity (⊥) directions). The morphology of the blends observed by confocal scanning laser microscopy and field emission scanning electron microscopy showed that under shearless conditions, starch-zein formed a co-continuous blend. The application of shear transformed this structure into a dispersion, with zein as the dispersed phase. The size of the zein aggregates increased with zein concentration. A certain degree of anisotropy was found with the 10% zein blend at the highest shear rate applied . Here, the zein aggregates were slightly deformed along the shear flow (∥). Both microscopy and tensile tests indicated that the blends have no or weak adhesion between the zein and starch phases. The effect of zein particles on the mechanical properties followed a modified model used for particle-matrix materials with weak adhesion between the phases.     

Identification of trinucleotide repeat ligands with a FRET melting assay

DNA hairpin structures formed within a repeated tract might be a causative factor for triplet expansion observed in several debilitating diseases. We have designed and used a fluorescence resonance energy transfer (FRET) melting assay to screen for ligands that bind specifically to the CNG triplet repeats. Using this assay, we screened a panel of 33 chemicals that were previously designed to bind DNA or RNA secondary structures. Remarkably, we found that macrocyclic compounds, such as acridine dimers and trimers, exhibit interesting affinities and specificities for this motif.     

The role of support on the performance of platinum-based catalysts for the total oxidation of polycyclic aromatic hydrocarbons

A range of Pt supported catalysts have been evaluated for the total oxidation of naphthalene. Catalysts contained 0.5 wt% Pt on a range of supports (γ-Al₂O₃, TiO₂, SiO₂, SnO₂, and CeO₂₎. SiO₂ was the best support, the 0.5%Pt/SiO₂ catalyst showing a conversion to carbon dioxide of over 90% at 200 °C (100 vppm naphthalene, GHSV = 45,000 h⁻¹). The catalyst also showed a considerably higher activity (in the temperature range 100–175 °C) than a CeO₂ catalyst recently reported to be one of the most effective catalysts for the total oxidation of naphthalene. The high activity of the 0.5%Pt/SiO₂ catalyst has been attributed to the relatively low dispersion and relatively large size of Pt particles. Furthermore, due to the acidic and non-reducible nature of the SiO₂, platinum is expected to have a weak interaction with the support. XPS data identified the presence of Pt⁰ on the surface and this contributes to the high activity.     

Effects of Interface Roughness on Residual Stresses in Thermal Barrier Coatings

Using a newly developed object-oriented finite-element analysis method, both an actual microstructure and model microstructures of a plasma-sprayed thermal barrier coating system were numerically simulated to analyze the full-field residual stresses of this coating system. Residual stresses in the actual microstructure were influenced by both the irregular top-coat/bond-coat interface and cracks in the top coat. By treating the microcracked top coat as a more-compliant solid microstructure, the effects of the irregular interface on residual stresses were examined. These results then could be compared to results that have been obtained by analyzing a model microstructure with a sinusoidal interface, which has been considered by some earlier investigators.     

Inhibition of Autophagy Does Not Re-Sensitize Acute Myeloid Leukemia Cells Resistant to Cytarabine

Elevated activation of the autophagy pathway is currently thought to be one of the survival mechanisms allowing therapy-resistant cancer cells to escape elimination, including for cytarabine (AraC)-resistant acute myeloid leukemia (AML) patients. Consequently, the use of autophagy inhibitors such as chloroquine (CQ) is being explored for the re-sensitization of AraC-resistant cells. In our study, no difference in the activity of the autophagy pathway was detected when comparing AraC-Res AML cell lines to parental AraC-sensitive AML cell lines. Furthermore, treatment with autophagy inhibitors CQ, 3-Methyladenine (3-MA), and bafilomycin A1 (BafA1) did not re-sensitize AraC-Res AML cell lines to AraC treatment. However, in parental AraC-sensitive AML cells, treatment with AraC did activate autophagy and, correspondingly, combination of AraC with autophagy inhibitors strongly reduced cell viability. Notably, the combination of these drugs also yielded the highest level of cell death in a panel of patient-derived AML samples even though not being additive. Furthermore, there was no difference in the cytotoxic effect of autophagy inhibition during AraC treatment in matched de novo and relapse samples with differential sensitivity to AraC. Thus, inhibition of autophagy may improve AraC efficacy in AML patients, but does not seem warranted for the treatment of AML patients that have relapsed with AraC-resistant disease.