Characterisation of two novel cyclodextrinases using on-line microdialysis sampling with high-performance anion exchange chromatography

In this work, a real-time sampling/analytical method for on-line measurements of two newly discovered cyclomaltodextrinases (CDases) has been developed and evaluated. This novel methodology not only allows the final products to be investigated, but it also reveals enzyme-specific differences in the degradation pathways during the hydrolysis of different substrates, which is a great advantage in the important tasks of investigating the mechanisms of and classifying new hydrolases, and is an advantage that conventional techniques cannot offer. Two different enzymes, one CDase from Laceyella sacchari (LsCda13) and one from Anoxybacillus flavithermus (AfCda13), were investigated during the hydrolysis of α-, β- and γ-cyclodextrin, and the hydrolysis products were sampled via a microdialysis probe and injected on-line every 30 min into a high-performance anion exchange chromatography system equipped with a pulsed amperometric detector (HPAEC–PAD), where they were identified. The enzymes yielded the same end-products, maltose and glucose, in an approximate molar ratio of 2:1, but they exhibited distinctly different patterns of intermediate product formation before reaching the end-point. LsCda13 had a more random distribution of the intermediate products, whereas AfCda13 showed the distinct intermediate production of maltotriose, which in some cases accumulated.     

An electron spectroscopic determination of theL I energy in the elements Sodium (Z=11) to Copper (Z=29)

Energies of atomic electrons in theK andL shells of eighteen elements of low atomic numbers have been measured by electron spectroscopy. Tabulated values for theL _I energies are shown to be in need of considerable revision. AluminumKα radiation is shown to have high potentialities for ESCA studies.     

Bounds on the CP asymmetry in like-sign dileptons from B(0)B*(0) meson decays

We have measured the charge asymmetry in like-sign dilepton yields from B(0)B*(0) meson decays using the CLEO detector at the Cornell Electron Storage Ring. We find a(0)(ll) identical with[N(l(+)l(+))-N(l(-)l(-))]/[N(l(+)l(+))+N(l(-)l(-))] = +0.013+/-0.050+/-0.005. We combine this result with a previous, independent measurement and obtain Re(epsilon(B))/(1+ the absolute value of epsilon(B)(2)) = +0.0035+/-0.0103+/-0.0015 (uncertainties are statistical and systematic, respectively) for the CP impurity parameter, epsilon(B).     

Aglycone specificity of <Emphasis Type="Italic">Thermotoga neapolitana</Emphasis> β-glucosidase 1A modified by mutagenesis,leading to increased catalytic efficiency in quercetin-3-glucoside hydrolysis

Background  

The thermostable β-glucosidase (TnBgl1A) from Thermotoga neapolitana is a promising biocatalyst for hydrolysis of glucosylated flavonoids and can be coupled to extraction methods using pressurized hot water. Hydrolysis has however been shown to be dependent on the position of the glucosylation on the flavonoid, and e.g. quercetin-3-glucoside (Q3) was hydrolysed slowly. A set of mutants of TnBgl1A were thus created to analyse the influence on the kinetic parameters using the model substrate para-nitrophenyl-β-D-glucopyranoside (pNPGlc), and screened for hydrolysis of Q3.     

Auger spectra for elements of low atomic number

KLL Auger spectra for the light elements sodium (Z=11) and magnesium (Z=12) have been studied. All five lines predicted by the extremeLS coupling theory are observed. A comparison is made with intensities predicted by the inter-configuration interaction theory ofAsaad. Agreement is obtained for the transition intensities to the three differentKLL configurations; however, theory does not give correct relative intensities within the 2s ² 2p ⁴ configuration. The measured Auger energies are compared with those calculated from the semi-empirical expression ofAsaad andBurhop and modified byHörnfeldt et al. Good agreement is obtained when we use electron binding energies measured by the ESCA method.