Comprehensive Analysis of Carbohydrate-Active Enzymes from the Filamentous Fungus Scytalidium candidum 3C.

Complete enzymatic degradation of plant polysaccharides is a result of combined action of various carbohydrate-active enzymes (CAZymes). In this paper, we demonstrate the potential of the filamentous fungus Scytalidium candidum 3C for processing of plant biomass. Structural annotation of the improved assembly of S. candidum 3C genome and functional annotation of CAZymes revealed putative gene sequences encoding such proteins. A total of 190 CAZyme-encoding genes were identified, including 104 glycoside hydrolases, 52 glycosyltransferases, 28 oxidative enzymes, and 6 carbohydrate esterases. In addition, 14 carbohydrate-binding modules were found. Glycoside hydrolases secreted during the growth of S. candidum 3C in three media were analyzed with a variety of substrates. Mass spectrometry analysis of the fungal culture liquid revealed the presence of peptides identical to 36 glycoside hydrolases, three proteins without known enzymatic function belonging to the same group of families, and 11 oxidative enzymes. The activity of endo-hemicellulases was determined using specially synthesized substrates in which the glycosidic bond between monosaccharide residues was replaced by a thio-linkage. During analysis of the CAZyme profile of S. candidum 3C, four ß-xylanases from the GH10 family and two ß-glucanases from the GH7 and GH55 families were detected, partially purified, and identified.

True perylene epitaxy on Ag(110) driven by site recognition effect.

We present a STM study of room temperature perylene adsorption on the Ag(110) surface. We have found a 2D perylene crystalline phase coexisting with the perylene liquid phase under thermal equilibrium. The reversible precipitation of the liquid phase at sub-monolayer coverage reveals the well ordered chiral crystalline phase existing in two enantiomorphic configurations of the ((-2)(3) (5)(2)) and ((2)(3) (5)(-2)) symmetry. This chiral phase is spatially separated into the 2D enantiopure islands of tens of nanometers size randomly distributed on the substrate and surrounded by the liquid medium. Analysis of surface registry of the crystalline phase combined with modeling of the intermolecular interactions indicates that its structure and symmetry is determined by a specific balance between the intermolecular attraction and intrinsic ability of the perylene aromatic board to recognize adsorption sites. The recognition effect was found to be strong enough to pin half of the perylene molecules into defined adsorption sites providing the structure skeleton. The attractive intermolecular interaction was found to be strong enough to bind another half of the molecules to the perylene skeleton shaping the true epitaxial structure.

Spontaneous cavitation in a Lennard-Jones liquid at negative pressures.

We report a molecular dynamics (MD) study of homogeneous bubble nucleation in a Lennard-Jones liquid under a negative pressure (cavitation). The rate of bubble nucleation has been determined in the range 2 x 10(-9) < J(*) = Jσ(4)(m/ε)(1/2) < 6 x 10(-6) by the mean lifetime method at temperatures T(*) = kBT/ε = 0.35, 0.4, 0.5, 0.6, 0.7, 0.8, 0.4, 0.5, 0.6, 0.7, 0.8. In molecular dynamics simulation calculations have also been made of the coefficient of bubble size diffusion, the Zeldovich nonequilibrium factor, and the radius of a critical nucleus R*. Different approaches to the determination of the nucleation rate in a stretched liquid have been considered in the framework of classical nucleation theory (CNT). The values of J obtained in MD simulation are by 8-20 orders higher than those predicted by CNT. The work of formation of a critical bubble and the dependence of surface tension γ(R*) at the critical bubble-liquid interface have been determined by data of MD simulation from CNT. The values of γ obtained have been approximated by an extended Tolman formula that takes into account, besides a linear correction, also the quadratic in curvature terms. The Tolman length δ∞ is negative and equals -(0.1-0.2)σ. The coefficient at 1/R*(2) is positive and does not exceed σ(2).

Improvement of the efficiency of transglycosylation catalyzed by α-galactosidase from Thermotoga maritima by protein engineering.

At high concentrations of p-nitrophenyl-α-D-galactopyranoside (pNPGal) as a substrate, its hydrolysis catalyzed by α-galactosidase from Thermotoga maritima (TmGalA) is accompanied by transglycosylation resulting in production of a mixture of (α1,2)-, (α1,3)-, and (α1,6)-p-nitrophenyl (pNP)-digalactosides. Molecular modeling of the reaction stage preceding the formation of the pNP-digalactosides within the active site of the enzyme revealed amino acid residues which modification was expected to increase the efficiency of transglycosylation. Upon the site-directed mutagenesis to the predicted substitutions of the amino acid residues, genes encoding the wild type TmGalA and its mutants were expressed in E. coli, and the corresponding enzymes were isolated and tested for the presence of the transglycosylating activity in synthesis of different pNP-digalactosides. Three mutants, F328A, P402D, and G385L, were shown to markedly increase the total transglycosylation as compared to the wild type enzyme. Moreover, the F328A mutant displayed an ability to produce a regio-isomer with the (α1,2)-bond at yield 16-times higher than the wild type TmGalA.

[Evaluation of regional microcirculation of the intestine in patients with inflammatory diseases of the uterus and its appendages complicated by diffuse peritonitis].

The microcirculation in the intestinal wall was evaluated in 42 women in the postoperative period using contact biomicroscopy in laparoscopy with a microhysteroscope. The dynamic contact biomicroscopy unlike other clinico-laboratory data allowed prognosis of the development or progress of peritonitis and choice of the strategy of treatment.

Atomic-scale imaging of insulating diamond through resonant electron injection.

The electronic properties of insulators such as diamond are of interest not only for their passive dielectric capabilities for use in electronic devices, but also for their strong electron confinement on atomic scales. However, the inherent lack of electrical conductivity in insulators usually prevents the investigation of their surfaces by atomic-scale characterization techniques such as scanning tunnelling microscopy (STM). And although atomic force microscopy could in principle be used, imaging diamond surfaces has not yet been possible. Here, we demonstrate that STM can be used in an unconventional resonant electron injection mode to image insulating diamond surfaces and to probe their electronic properties at the atomic scale. Our results reveal striking electronic features in high-purity diamond single crystals, such as the existence of one-dimensional fully delocalized electronic states and a very long diffusion length for conduction-band electrons. We expect that our method can be applied to investigate the electronic properties of other insulating materials and so help in the design of atomic-scale electronic devices.

Electronic structure of partially hydrogenated Si(100)-( 2 x 1) surfaces prepared by thermal and nonthermal desorption.

The electronic structure of partially hydrogenated Si(100)- (2 x 1) surfaces, prepared by controlled thermal annealing and nonthermal photon stimulated desorption of fully hydrogenated Si(100) surfaces, has been investigated by using valence band photoemission. Thermal and nonthermal desorption are found to produce very specific electronic surface structures. This led us to the discovery of two specific surface states having binding energies of 1.0 and 0.7 eV associated with the isolated Si dimers and single Si dangling bonds, respectively.