Bioelectrochemical detection of L-lactate respiration using genetically modified Hansenula polymorpha yeast cells overexpressing flavocytochrome b2

In general, L-lactate respiration is difficult to detect in living yeast cells due to the small activity of L-lactate oxidizing enzymes within the mitochondria. Genetically modified cells of methylotrophic yeast Hansenula polymorpha overproducing L-lactate:cytochrome c-oxidoreductase (EC 1.1.2.3, also known as flavocytochrome b₂, FC b₂) were physically immobilized by means of a dialysis membrane onto various types of electrode materials in order to investigate the possibility of electrochemically detecting L-lactate respiration. It could be shown that in the case of genetically modified Hansenula polymorpha cells in contrast to cells from the parental strain, enhanced L-lactate-dependent respiration could be detected. Due to overproduction of FC b₂ the O₂ reduction current is decreased upon addition of L-lactate to the electrolyte solution. The electron transfer pathway in the L-lactate-dependent respiration process involves a cascade over three redox proteins, FC b₂, cytochrome c and Complex-IV, starting with L-lactate oxidation and ending with oxygen reduction. By means of selective inhibition of Complex IV with CN⁻, lactate respiration could be proven for causing the decrease in the O₂ reduction.     

Novel design of multicapillary arrays for high-throughput DNA sequencing

A novel approach to design and optimize linear multicapillary arrays (LMCAs) for high-throughput DNA sequencing is proposed. A significant increase in the number of capillary lanes is obtained due to the use of composite insertions alternately placed between working capillaries of the array and a specific combination of refractive indices of the DNA separation matrix, capillary glass, the insertions and a medium which surrounds the capillary array. Theoretical and experimental studies showed that in conjunction with a dual-side laser illumination scheme, the proposed LMCA design allows a simultaneous uniform irradiation of as many as 550 working capillaries.     

Invertible architectures from amphiphilic polyesters

We synthesized and characterized novel amphiphilic polyesters with both hydrophilic and hydrophobic functionalities. The polyesters are soluble in organic and aqueous media and reveal the formation of inverse architectures whose behavior could be correlated to their chemical structure. We foresee that the amphiphilic properties of the polyesters reported here are obviously the basis of new architectures both in solution and on the solid surfaces, which could be used in a broad range of applications. The described synthesis of the copolymers is very simple and is based on commercially available products. That makes this approach attractive in various uses.     

Biosynthesis of the terpene phenalinolactone in Streptomyces sp. Tü6071: analysis of the gene cluster and generation of derivatives

Phenalinolactones are terpene glycosides with antibacterial activity. A striking structural feature is a highly oxidized gamma-butyrolactone of elusive biosynthetic origin. To investigate the genetic basis of the phenalinolactones biosynthesis, we cloned and sequenced the corresponding gene cluster from the producer strain Streptomyces sp. Tü6071. Spanning a 42 kbp region, 35 candidate genes could be assigned to putatively encode biosynthetic, regulatory, and resistance-conferring functions. Targeted gene inactivations were carried out to specifically manipulate the phenalinolactones pathway. The inactivation of a sugar methyltransferase gene and a cytochrome P450 monoxygenase gene led to the production of modified phenalinolactone derivatives. The inactivation of a Fe(II)/alpha-ketoglutarate-dependent dioxygenase gene disrupted the biosynthetic pathway within gamma-butyrolactone formation. The structure elucidation of the accumulating intermediate indicated that pyruvate is the biosynthetic precursor of the gamma butyrolactone moiety.     

The role of capillary and surface forces in the crossover behavior of solid nanoparticles at liquid interfaces

We investigate the interaction between a nanoparticle and an oil-water interface with particular emphasis on the particle crossing through the interface. The formation of a three-phase contact line is investigated in two cases, namely in the presence and in the absence of surface forces. We carefully examine the interplay between capillary and surface forces in such systems. Two instabilities of the interface (snap-in/snap-out) as the particle is moved through the interface are identified and quantitatively described. While the snap-in instability was observed in some AFM studies, the precise interface position and configuration relative to the particle at the instability depends on the nature of the surface forces present in the system. After the snap-in, the particle is adsorbed and must overcome an energy barrier due to the interface deformation in order to cross-over to the other liquid. We make quantitative predictions on the interface configuration at the instabilities and the free energy barrier height. The roles of particle size and different interaction parameters characterizing the system in determining the magnitude of the energy barrier for crossing and in the formation of a three-phase contact line are discussed. Ultimately, this study will enable us to make quantitative predictions on capillary effects in nanoparticle-microemulsions mixtures and other colloidal systems. For particles in the micrometer range and larger the capillary forces dominate over the surface forces and dictate how the snap-in occurs. However, the situation becomes different for particle sizes smaller than about 100 nm. The presence of surface forces modifies the interface configuration and the free energy jump at the snap-in instability.     

Spatial hydration structures and dynamics of phenol in sub- and supercritical water

The hydration structures and dynamics of phenol in aqueous solution at infinite dilution are investigated using molecular-dynamics simulation technique. The simulations are performed at several temperatures along the coexistence curve of water up to the critical point, and above the critical point with density fixed at 0.3 g/cm3. The hydration structures of phenol are characterized using the radial, cylindrical, and spatial distribution functions. In particular, full spatial maps of local atomic (solvent) density around a solute molecule are presented. It is demonstrated that in addition to normal H bonds with hydroxyl group of phenol, water forms pi-type complexes with the center of the benzene ring, in which H2O molecules act as H-bond donor. At ambient conditions phenol is solvated by 38 water molecules, which make up a large hydrophobic cavity, and forms on average 2.39 H bonds (1.55 of which are due to the hydroxyl group-water interactions and 0.84 are due to the pi complex) with its hydration shell. As temperature increases, the hydration structure of phenol undergoes significant changes. The disappearance of the pi-type H bonding is observed near the critical point. Self-diffusion coefficients of water and phenol are also calculated. Dramatic increase in the diffusivity of phenol in aqueous solution is observed near the critical point of simple point-charge-extended water and is related to the changes in water structure at these conditions.     

Calix[4]arene alpha-aminophosphonic acids: asymmetric synthesis and enantioselective inhibition of an alkaline phosphatase

[structure: see text] Chiral calix[4]arene alpha-aminophosphonic acids were obtained through diastereoselective Pudovik-type addition of sodium ethyl phosphites to the chiral calixarene imines, removal of chiral auxiliary groups, and mild dealkylation of phosphonate fragments. The diacids obtained show inhibitory activity toward porcine kidney alkaline phosphatase that depends considerably on the absolute configuration of the alpha-carbon atoms.     

Algebraic Nijenhuis operators and Kronecker Poisson pencils

We give a criterion of (micro-)kroneckerity of the linear Poisson pencil on g^∗ related to an algebraic Nijenhuis operator on a finite-dimensional Lie algebra g. As an application we get a series of examples of completely integrable systems on semisimple Lie algebras related to Borel subalgebras and a new proof of the complete integrability of the free rigid body system on gln.     

Tandem intramolecular Michael-aldol reaction as a tool for the construction of the C-ring of hexacyclinic acid

[reaction: see text] The tandem intramolecular Michael-aldol reaction was studied as a tool for the construction of the C-ring of hexacyclinic acid. By changing the reaction conditions it was possible to selectively obtain either the kinetic or the thermodynamic product. Retro-aldol reaction and subsequent epimerization provides four individual cyclopentane derivatives that can be incorporated as building blocks in natural product syntheses.     

Two-color vector solitons in nonlocal media

We investigate the interaction between two beams differing in wavelength and the properties of dual-frequency spatial solitons in nonlocal birefringent reorientational media. We report the first experimental observations of anisotropic nonlocal vector solitons in unbiased nematic liquid crystals. Model and simulations, based on the paraxiality along the Poynting vectors, include joint walk-off and breathing.