Kinetics of the adsorption of bovine serum albumin contained in a model wine solution by non-swelling ion-exchange resins

Adsorption of wine proteins is an essential step in the production of white and rosé wines. In order to develop environmentally friendly adsorption processes, non-swelling adsorbents are required. The performance of selected non-swelling ion-exchange resins (Macro-Prep™ 50S and Streamline® SP) was studied by describing the process kinetics of the adsorption of BSA in a model wine solution. The process was assumed to be diffusion controlled and a shrinking core model was applied. Experiments were performed in the 5–35°C temperature range and with different equilibrium partition coefficients. The results obtained with the shrinking core model were theoretically consistent and the apparent diffusivity values correlated very well with theoretically estimated effective diffusivities combined with a linear dependence of porosity with temperature. Separating the temperature effect on porosity, the apparent diffusivity followed an Arrhenius type dependency with temperature with 16.9 kJ/mole activation energy

Controlled Orientation of Active Sites in a Nanostructured Multienzyme Complex

Multistep cascade reactions in nature maximize reaction efficiency by co-assembling related enzymes. Such organization facilitates the processing of intermediates by downstream enzymes. Previously, the studies on multienzyme nanocomplexes assembled on DNA scaffolds demonstrated that closer interenzyme distance enhances the overall reaction efficiency. However, it remains unknown how the active site orientation controlled at nanoscale can have an effect on multienzyme reaction. Here, we show that controlled alignment of active sites promotes the multienzyme reaction efficiency. By genetic incorporation of a non-natural amino acid and two compatible bioorthogonal chemistries, we conjugated mannitol dehydrogenase to formate dehydrogenase with the defined active site arrangement with the residue-level accuracy. The study revealed that the multienzyme complex with the active sites directed towards each other exhibits four-fold higher relative efficiency enhancement in the cascade reaction and produces 60% more D-mannitol than the other complex with active sites directed away from each other.ope

Im Rennen um die besten Köpfe

IM RENNEN UM DIE BESTEN KÖPFE Im Rennen um die besten Köpfe (1) Inhaltsverzeichnis (4) 1 Einleitung (6) 1.1 Problemstellung (6) 1.2 Zielsetzung und Nutzen dieser Arbeit (8) 1.3 Forschungsfrage (9) 1.4 Aufbau der Arbeit (10) 2 Forschungsstand (10) 2.1 Erwartungen der Arbeitnehmer*innen (11) 2.2 Inhalte der Stellenausschreibungen und Hypothesen (13) 2.2.1 Gehalt (13) 2.2.2 Art des Unternehmens (14) 2.2.3 Work-Life-Balance (15) 2.2.4 Gestaltung der Stellenausschreibung (17) 3 Empirie (18) 3.1 Forschungsfeld (19) 3.2 Methodische Herangehensweise (Conjoint Analyse) (19) 3.2.1 Auswahl von Eigenschaften und Eigenschaftsausprägungen (20) 3.2.2 Erhebungsdesign (22) 3.2.3 Bewertung der Stimuli (23) 3.2.4 Schätzung der Nutzenfunktion (24) 3.3 Operationalisierung & zu erhebenden Variablen (25) 3.4 Datenerhebung (30) 3.5 Beschreibung der Stichprobe (30) 3.6 Datenauswertung (32) 3.7 Forschungsethik/Gütekriterien (33) 3.8 Ergebnis (34) 4 Diskussion (40) 4.1 Zusammenfassung und Beantwortung der Forschungsfrage (40) 4.2 Rückführung zur Theorie (42) 4.3 Handlungsempfehlungen (44) 4.4 Limitationen und Forschungsausblick (45) Literaturverzeichnis (47) Hilfsmittelverzeichnis (55) Abbildungs- und Tabellenverzeichnis (55) Abkürzungsverzeichnis (55) Anhang (57) Umfrage (57) R-Code (61

Influence of intrinsic factors on conventional wine protein stability tests

The influence of intrinsic factors on the results of ethanol, tannin and heat tests, routinely used to assess wine protein stability, was studied. Experiments were performed on 23 Portuguese and Austrian wines. The factors considered were total protein content, pH, ethanol content and the amount of several relevant cations (calcium, iron, copper, sodium and potassium). The protein pro®les were analysed by HPLC fractionation. The heat test was a good indicator of total protein content while the ethanol and the tannin tests showed signi®cant interference by the other factors. A factorial design at two levels in selected samples was also performed to assess the influence of pH, storage temperature, tannin concentration and ethanol concentration on the development of turbidity. Results indicated that ethanol content had no significant infuence, and that pH and storage temperature had a significant infuence, though only when tannin was added

Kinetic Study of the Catalytic Mechanism of Mannitol Dehydrogenase from <i>Pseudomonas fluorescens</i>^†

To characterize catalysis by NAD-dependent long-chain mannitol 2-dehydrogenases (MDHs), the recombinant wild-type MDH from Pseudomonas fluorescens was overexpressed in Escherichia coli and purified. The enzyme is a functional monomer of 54 kDa, which does not contain Zn2+ and has B-type stereospecificity with respect to hydride transfer from NADH. Analysis of initial velocity patterns together with product and substrate inhibition patterns and comparison of primary deuterium isotope effects on the apparent kinetic parameters, Dkcat, D(kcat/KNADH), and D(kcat/Kfructose), show that MDH has an ordered kinetic mechanism at pH 8.2 in which NADH adds before d-fructose, and d-mannitol and NAD are released in that order. Isomerization of E−NAD to a form which interacts with d-mannitol nonproductively or dissociation of NAD from the binary complex after isomerization is the slowest step (≥110 s-1) in d-fructose reduction at pH 8.2. Release of NADH from E−NADH (32 s-1) is the major rate-limiting step in mannitol oxidation at this pH. At the pH optimum for d-fructose reduction (pH 7.0), the rate of hydride transfer contributes significantly to rate limitation of the catalytic cascade and the overall reaction. D(kcat/Kfructose) decreases from 2.57 at pH 7.0 to a value of ≤1 above pH 9.6, corresponding to the pK of 9.34 observed in the pH profile of kcat/Kfructose. Therefore, hydride transfer is not pH-dependent, and d-fructose is not sticky at pH 7.0. A comparison of the kinetic data of MDH and mammalian sorbitol dehydrogenase, presumably involved in detoxification metabolism, is used to point out a physiological function of MDH in the oxidation of d-mannitol with high specificity and fluxional efficiency under prevailing reaction conditions in vivo