Rationalizing perhydrolase activity of aryl-esterase and subtilisin Carlsberg mutants by molecular dynamics simulations of the second tetrahedral intermediate state

The perhydrolysis reaction in hydrolases is an important example of catalytic promiscuity and has many potential industrial applications. The mechanisms of perhydrolase activity of a subtilisin Carlsberg mutant and of an aryl-esterase mutant have been investigated using classical molecular dynamics simulations of the second tetrahedral intermediate (TI) state. The simulations demonstrated that hydrogen bonding between the second TI of the perhydrolysis reaction is possible in the mutants but not wild type. The stabilization by hydrogen bonds was specific for the perhydrolysis intermediate and either no hydrogen bonding or only weakened hydrogen bonding to the second TI state of the hydrolysis reaction was observed. Furthermore, a significant hindrance to the formation of the catalytically important hydrogen bond between His64 and Ser221 in the catalytic triad by competing hydrogen bonds was found for the subtilisin mutant but not wild type enzyme in case of the hydrolysis intermediate. The opposite was observed in case of the perhydrolysis intermediate. The result offers a qualitative explanation for the overall reduced hydrolysis activity of the subtilisin mutant. In addition, the simulations also explain qualitatively the perhydrolysis activity of the enzyme variants and may be helpful for designing enzyme mutants with further improved perhydrolysis activity.     

Quantification of (−)-Epicatechin and Procyanidin B2 in Chocolates

JPC – Journal of Planar Chromatography – Modern TLC - Validation of a method for the quantification of (−)-epicatechin and procyanidin B2 in chocolates is reported. After a simple...     

Sturm theorems for second order linear nonhomogenous differential equations and localization of zeros of the solution

It is shown that Sturm theorems, formulated in the 1830??s ([1], [2], [3] and [4]) and valid for second order linear homogeneous differential equation L(y)??y??+a(x)y??+b(x)y=0, could as well be formulated for the class of nonhomogeneous linear differential equations L(y)=f(x). Criteria for the existence of oscillatory solutions of nonhomogeneous equations, as well as more exact locations of the zeros are given.     

New results on (b,c)–inverses

In this paper, we present some necessary and sufficient conditions for the existence of the (b, c)-inverse and several representations for the (b, c)-inverse related to the group inverse. Since Mary inverse, core inverse, dual core inverse and Bott–Duffin (e, f)-inverse are all the special cases of the (b, c)-inverse, related results for these inverses are obtained.     

A NOTE ON THE DRAZIN INVERSE OF A MODIFIED MATRIX

In this article,the expression for the Drazin inverse of a modified matrix is considered and some interesting results are established.This contributes to certain recent results obtained by Y.Wei [9].     

Dielectric response of fibrous polyethyleneterephtalate

The effect of the electric periodic field on the dielectric parameters (relative dielectric permeability and dielectric tangent of losses) of fibrous polyethyleneterephtalate (PET) different structures was studied. The analysis of dielectric relaxation spectra has been performed in the frequency range from 90 kHz to 3 MHz, while the sample was heating from 120 to 355 K. In this range of measurements, β relaxation is observed and the activation energy of molecular motion is established. Through a comparison of obtained results, a relationship is noted between dielectric parameters and structural properties of a sample. It is observed that samples whose yarn is obtained from short fibre have lower value of dielectric parameters than samples with monofilament and multifilament yarn. Samples with higher values of volume fraction of fibres show higher values of dielectric parameters. In room conditions a comparison between dielectric parameters of the samples and a sample with incorporated electrically conductive threads has been performed. The dielectric response of the tested fibrous PET is comparable with the dielectric response of other PET – based structures.     

Calorimetric investigation of Al–Zn alloys using Oelsen method

The results of calorimetric study of binary Al–Zn system done according to the Oelsen thermodynamic method are presented in this paper. Main thermodynamic properties, including activities, activity coefficients, partial/integral molar Gibbs excess, and mixing energies were determined at 1,000 K. Positive deviation from Raoult law was noticed, with minimal values of ΔG ^M about ?3 kJ mol^?1 and maximal values of ΔG ^E about +2 kJ mol^?1. The energetics of mixing in liquid Al–Zn alloys has been analyzed through the study of concentration fluctuation in the long-wavelength limit, and weak affinity between Al and Zn atoms in the system was observed. Differential thermal analysis and light optic microscopy were applied in the frame of characterization of investigated binary alloys and phase diagram examination, and the results obtained were in accordance with available literature data.     

Determination of multi-class pharmaceuticals in wastewater by liquid chromatography–tandem mass spectrometry (LC–MS–MS)

An analytical method for multi-class pharmaceuticals determination in wastewater has been developed and validated. Target compounds were: sulfonamides (sulfadiazine, sulfaguanidine, sulfamethazine, sulfamethoxazole), fluoroquinolones (ciprofloxacin, enrofloxacin, norfloxacin), diaminopyrimidine (trimethoprim), anaesthetic (procaine), anthelmintic (praziquantel and febantel), and macrolide (roxithromycin). The method involves pre-concentration and clean-up by solid-phase extraction (SPE) using Strata-X extraction cartridges at pH 4.0. Target analytes were identified and quantitatively determined by liquid chromatography–tandem mass spectrometry using multiple reaction monitoring (MRM). Recoveries were higher than 50% with relative standard deviation (RSD) below 18.3% for three concentrations. Only for sulfaguanidine was low recovery obtained. Matrix effect was evaluated using matrix-matched standards. The method detection limit (MDL) was between 0.5 and 5 ng L⁻¹ in spiked water samples. The precision of the method, calculated as relative standard deviation, ranged from 0.5 to 2.0% and from 1.4 to 8.3 for intra-day and inter-day analysis, respectively. The described analytical method was used for determination of pharmaceuticals in effluent wastewaters from the pharmaceutical industry.