Surface characterization and interfacial activity of chitinase chi18-5 against chitosan in langmuir monolayers.

One of the challenges for producing active chitinase formulations relies on the gap between the laboratory tests and the biological scenarios where the enzyme will perform its function. In this work, we have employed different Langmuir monolayer arrays to evaluate the interfacial behavior of a recently purified recombinant chitinase, Chi18-5. We have demonstrated that two conformations exist for the chitinase at pH values close to its pI, showing very distinct structural properties at the air/aqueous interface. Enzyme activity was assessed by implementing different kinetic approaches and using a chitosan-1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) mixed film as organized substrate model membrane. Combining these strategies, we demonstrated that better catalytic efficiencies can be obtained for Chi18-5 at pH 5. Moreover, the chitinase activity at the air/aqueous interface can be tuned by introducing in situ pH modifications over the surrounding milieu. We also studied the changes in the topography at the mesoscale level using Brewster Angle Microscopy (BAM). We found that Chi18-5 segregated onto the chitosan domains of the membrane, showing differences in homogeneity depending on the pH imposed. Alternatively, pure Chi18-5 was tested for immobilization onto a hydrophilic activated solid support using the Langmuir-Blodgett technique. Atomic Force Microscopy (AFM) analyses showed successfully stabilization and preservation of molecular features attributed to the pH at which the enzyme deposition was performed.

Uniaxial and biaxial structures in the elastic Maier-Saupe model.

We perform statistical mechanics calculations to analyze the global phase diagram of a fully connected version of a Maier-Saupe-Zwanzig lattice model with the inclusion of couplings to an elastic strain field. We point out the presence of uniaxial and biaxial nematic structures, depending on temperature T and on the applied stress σ. Under uniaxial extensive tension, applied stress favors uniaxial orientation, and we obtain a first-order boundary along which there is a coexistence of two uniaxial paranematic phases, and which ends at a simple critical point. Under uniaxial compressive tension, stress favors biaxial orientation; for small values of the coupling parameters, the first-order boundary ends at a tricritical point, beyond which there is a continuous transition between a paranematic and a biaxially ordered structure. For some representative choices of the model parameters, we obtain a number of analytic results, including the location of critical and tricritical points and the line of stability of the biaxial phase.

Binding of the substrate UDP-glucuronic acid induces conformational changes in the xanthan gum glucuronosyltransferase.

GumK is a membrane-associated glucuronosyltransferase of Xanthomonas campestris that is involved in xanthan gum biosynthesis. GumK belongs to the inverting GT-B superfamily and catalyzes the transfer of a glucuronic acid (GlcA) residue from uridine diphosphate (UDP)-GlcA (UDP-GlcA) to a lipid-PP-trisaccharide embedded in the membrane of the bacteria. The structure of GumK was previously described in its apo- and UDP-bound forms, with no significant conformational differences being observed. Here, we study the behavior of GumK toward its donor substrate UDP-GlcA. Turbidity measurements revealed that the interaction of GumK with UDP-GlcA produces aggregation of protein molecules under specific conditions. Moreover, limited proteolysis assays demonstrated protection of enzymatic digestion when UDP-GlcA is present, and this protection is promoted by substrate binding. Circular dichroism spectroscopy also revealed changes in the GumK tertiary structure after UDP-GlcA addition. According to the obtained emission fluorescence results, we suggest the possibility of exposure of hydrophobic residues upon UDP-GlcA binding. We present in silico-built models of GumK complexed with UDP-GlcA as well as its analogs UDP-glucose and UDP-galacturonic acid. Through molecular dynamics simulations, we also show that a relative movement between the domains appears to be specific and to be triggered by UDP-GlcA. The results presented here strongly suggest that GumK undergoes a conformational change upon donor substrate binding, likely bringing the two Rossmann fold domains closer together and triggering a change in the N-terminal domain, with consequent generation of the acceptor substrate binding site.

Maier-Saupe model for a mixture of uniaxial and biaxial molecules.

We introduce shape variations in a liquid-crystalline system by considering an elementary Maier-Saupe lattice model for a mixture of uniaxial and biaxial molecules. Shape variables are treated in the annealed (thermalized) limit. We analyze the thermodynamic properties of this system in terms of temperature T, concentration c of intrinsically biaxial molecules, and a parameter Δ associated with the degree of biaxiality of the molecules. At the mean-field level, we use standard techniques of statistical mechanics to draw global phase diagrams, which are shown to display a rich structure, including uniaxial and biaxial nematic phases, a reentrant ordered region, and many distinct multicritical points. Also, we use the formalism to write an expansion of the free energy in order to make contact with the Landau-de Gennes theory of nematic phase transitions.

Annealed Ising model with site dilution on self-similar structures.

We consider an Ising model on the triangular Apollonian network (AN), with a thermalized distribution of vacant sites. The statistical problem is formulated in a grand canonical ensemble, in terms of the temperature T and a chemical potential µ associated with the concentration of active magnetic sites. We use a well-known transfer-matrix method, with a number of adaptations, to write recursion relations between successive generations of this hierarchical structure. We also investigate the analogous model on the diamond hierarchical lattice (DHL). From the numerical analysis of the recursion relations, we obtain various thermodynamic quantities. In the µâ†’∞ limit, we reproduce the results for the uniform models: in the AN, the system is magnetically ordered at all temperatures, while in the DHL there is a ferromagnetic-paramagnetic transition at a finite value of T. Magnetic ordering, however, is shown to disappear for sufficiently large negative values of the chemical potential.

Biophysical characterization of the outer membrane polysaccharide export protein and the polysaccharide co-polymerase protein from Xanthomonas campestris.

This study investigated the structural and biophysical characteristics of GumB and GumC, two Xanthomonas campestris membrane proteins that are involved in xanthan biosynthesis. Xanthan is an exopolysaccharide that is thought to be a virulence factor that contributes to bacterial in planta growth. It also is one of the most important industrial biopolymers. The first steps of xanthan biosynthesis are well understood, but the polymerization and export mechanisms remain unclear. For this reason, the key proteins must be characterized to better understand these processes. Here we characterized, by biochemical and biophysical techniques, GumB, the outer membrane polysaccharide export protein, and GumC, the polysaccharide co-polymerase protein of the xanthan biosynthesis system. Our results suggested that recombinant GumB is a tetrameric protein in solution. On the other hand, we observed that both native and recombinant GumC present oligomeric conformation consistent with dimers and higher-order oligomers. The transmembrane segments of GumC are required for GumC expression and/or stability. These initial results provide a starting point for additional studies that will clarify the roles of GumB and GumC in the xanthan polymerization and export processes and further elucidate their functions and mechanisms of action.

Biaxial nematic phase in the Maier-Saupe model for a mixture of discs and cylinders.

We analyze the global phase diagram of a Maier-Saupe lattice model with the inclusion of shape-disordered degrees of freedom to mimic a mixture of oblate and prolate molecules (discs and cylinders). In the neighborhood of a Landau multicritical point, solutions of the statistical problem can be written as a Landau-de Gennes expansion for the free energy. If the shape-disordered degrees of freedom are quenched, we confirm the existence of a biaxial nematic structure. If orientational and disorder degrees of freedom are allowed to thermalize, this biaxial solution becomes thermodynamically unstable. Also, we use a two-temperature formalism to mimic the presence of two distinct relaxation times, and show that a slight departure from complete thermalization is enough to stabilize a biaxial nematic phase.

Tricritical-like behavior of the nonlinear optical refraction at the nematic-isotropic transition in the E7 thermotropic liquid crystal.

We use Z-scan technique to investigate the nonlinear optical response of the thermotropic liquid crystal E7 in the neighborhood of the nematic-isotropic phase transition. The analysis of the data for the nonlinear optical birefringence is compatible with an effective critical exponent of the order parameter, ß = 0.28 ± 0.03, which is close to the classical value, ß = 0.25 , for a tricritical point. The nonlinear optical absorption in the nematic range depends on the geometrical configuration of the nematic director with respect to the polarization beam, and vanishes in the isotropic phase.

Phase diagram of a model for a binary mixture of nematic molecules on a Bethe lattice.

We investigate the phase diagram of a discrete version of the Maier-Saupe model with the inclusion of additional degrees of freedom to mimic a distribution of rodlike and disklike molecules. Solutions of this problem on a Bethe lattice come from the analysis of the fixed points of a set of nonlinear recursion relations. Besides the fixed points associated with isotropic and uniaxial nematic structures, there is also a fixed point associated with a biaxial nematic structure. Due to the existence of large overlaps of the stability regions, we resorted to a scheme to calculate the free energy of these structures deep in the interior of a large Cayley tree. Both thermodynamic and dynamic-stability analyses rule out the presence of a biaxial phase, in qualitative agreement with previous mean-field results.

Statistical models of mixtures with a biaxial nematic phase.

We consider a simple Maier-Saupe statistical model with the inclusion of disorder degrees of freedom to mimic the phase diagram of a mixture of rodlike and disklike molecules. A quenched distribution of shapes leads to a phase diagram with two uniaxial and a biaxial nematic structure. A thermalized distribution, however, which is more adequate to liquid mixtures, precludes the stability of this biaxial phase. We then use a two-temperature formalism, and assume a separation of relaxation times, to show that a partial degree of annealing is already sufficient to stabilize a biaxial nematic structure.

Fluctuation-dissipation theorem and the linear Glauber model.

We obtain exact expressions for the two-time autocorrelation and response functions of the -dimensional linear Glauber model. Although this linear model does not obey detailed balance in dimensions d > or = 2, we show that the usual form of the fluctuation-dissipation ratio still holds in the stationary regime. In the transient regime, we show the occurrence of aging, with a special limit of the fluctuation-dissipation ratio, x(infinity) = 1/2, for a quench at the critical point.

Correlated disordered interactions on Potts models.

Using a weak-disorder scheme and real-space renormalization-group techniques, we obtain analytical results for the critical behavior of various q-state Potts models with correlated disordered exchange interactions along d(1) of d spatial dimensions on hierarchical (Migdal-Kadanoff) lattices. Our results indicate qualitative differences between the cases d-d(1)=1 (for which we find non-physical random fixed points, suggesting the existence of non-perturbative fixed distributions) and d-d(1)>1 (for which we do find acceptable perturbative random fixed points), in agreement with previous numerical calculations by Andelman and Aharony [Phys. Rev. B 31, 4305 (1985)]. We also rederive a criterion for relevance of correlated disorder, which generalizes the usual Harris criterion.