Friedrich Ruf

Adsorption and separation of proteins by a synthetic hydrotalcite.

In this study, the potential use of a synthetic Mg/Al hydrotalcite (layered double hydroxide) as a novel chromatography material for protein purification was investigated. The hydrotalcite is present in its carbonate form and is characterized by an Al/Mg-ratio of 1.85. Zetapotential measurements confirm a positive surface potential up to pH 10 suggesting applicability as anion exchanger. The binding of model proteins covering a broad range of isoelectric points and molecular weights was performed at different pH-values under batch conditions to evaluate the binding behaviour of the hydrotalcite. Furthermore, static binding capacities were exemplarily determined for hemoglobin and human serum albumin. Additionally, the adsorption and elution of hemoglobin was studied under dynamic conditions. The binding behaviour of the hydrotalcite was compared to commercially available anion exchangers and was found to be a function of pH, depending on the model protein. Variant adsorption behaviour is explained by further interactions like hydrogen bonds and by an unequal charge distribution over the protein surfaces. The hydrotalcite reveals high adsorption capacities under static (260 mg/g) as well as under dynamic conditions (88 mg/g at 34 cm/h; 61 mg/g at 340 cm/h). With appropriate buffers like 500 mM carbonate (pH 10) the adsorbed proteins can be nearly completely desorbed making regeneration possible. Due to the binding and elution properties it is concluded, that the hydrotalcite can serve anion exchange material for chromatographic protein separations.

Separation of patatins and protease inhibitors from potato fruit juice with clay minerals as cation exchangers

Potato fruit juice as a by-product of the starch industry contains proteins with interesting functionalities such as protease inhibitors or patatin with its high nutritional value. Due to their functional properties, these proteins are principally of industrial interest. A drawback for the application of these potato proteins is the separation and isolation under maintenance of the biological activity. So far, there are no methods in literature, which are satisfying concerning the costs or the separation performance. In this study, we show a chromatographic approach using natural clay minerals as cation exchangers to separate two protein fractions in potato fruit juice. Additionally, the content of glycoalkaloids naturally occurring in potatoes is significantly reduced in a single step together with the separation of the patatins and the protease inhibitors.

New zeolite adsorbents for downstream processing of polyphenols from renewable resources

Commercial materials with polyvinylpolypyrrolidone and polymeric amberlites (XAD7HP, XAD16) are commonly used for the adsorptive downstream processing of polyphenols from renewable resources. In this study, beta‐zeolite‐based adsorbent systems were examined, and their properties were compared to organic resins. Batch adsorption experiments were conducted with synthetic solutions of major polyphenols. Adsorption isotherms and desorption characteristics of individual adsorbent were determined based on these results. Maximum adsorption capacities were calculated using the Langmuir model. For example, the zeolites had capacities up to 203.2 mg/g for ferulic acid. To extend these results to a complex system, additional experiments were performed on rapeseed meal and wheat seed extracts as representative renewable resources. HPLC analysis showed that with 7.5% w/v, which is regarded as the optimum amount of zeolites, zeolites A and B could bind 100% of the major polyphenols as well as release polyphenols at high yields. Additionally, regeneration experiments were performed with isopropyl alcohol at 99°C to evaluate how zeolites regenerate under mild conditions. The results showed only a negligible loss of adsorption capacity and no loss of desorption capacity. In summary, it was concluded that beta‐zeolites were promising adsorbents for developing new processes to isolate polyphenols from renewable resources.

Recycling of spent oil bleaching earth as source of glycerol for the anaerobic production of acetone, butanol, and ethanol with Clostridium diolis and lipolytic Clostridium lundense

A major part of edible oil is subjected to bleaching procedures, primarily with minerals applied as adsorbers. Their recycling is currently done either by regaining the oil via organic solvent extraction or by using the spent bleaching earth (SBE) as additive for animal feed, etc. As a new method, the reutilization of the by‐product SBE for the microbiologic formation of acetone, butanol, and ethanol (ABE) is presented as proof‐of‐concept. The SBE was taken from a palm oil cleaning process. The recycling concept is based on the application of lipolytic clostridia strains. Due to considerably long fermentation times, co‐fermentation with Candida rugosa and enzymatic hydrolyses of the bound oil with a subsequent clostridia fermentation are shown as alternative routes. Anaerobic fermentations under comparison of different clostridia strains were performed with glycerol media, enzymatically hydrolyzed palm oil and SBE. Solutes, side product compositions and productivities were quantified via HPLC. A successful production of ABE solutes from SBE has been done with a yield of 0.15 g butanol per gram of bound glycerol. Thus, the biotechnological recycling of the waste stream is possible in principle. Inhibition of the substrate suspension has been observed. A chromatographic ion‐exchange of substrates increased the biomass concentration.

Layer‐by‐Layer‐Oberflächenfunktionalisierung für Chromatographiematerialien – eine preiswerte Alternative zu bestehenden Verfahren?

Die Abscheidung von gasgetragenen Nanopartikeln kann zum Aufbau funktioneller Partikelschichten, die in Katalyse, Solarzellen und Sensorik angewandt werden, eingesetzt werden. Dabei bestimmen Bewegung und Morphologie der Ausgangspartikeln die Struktur der Partikelschicht (u. a. Porositat, Koordinationszahl, fraktale Dimension) [1]. Durch die Variation der KnudsenZahl kann die Bewegung der Nanopartikeln von diffusiv bis ballistisch eingestellt werden. Zudem erhoht die bei hinreichend grosen Impaktionsgeschwindigkeiten beobachtete Abplattung der Nanopartikel-Agglomerate die Packungsdichte der Partikelschicht [2]. Silber-Nanopartikeln wurden durch einen Verdampfung-Kondensation-Prozess in einem Rohrofen hergestellt und anschliesend in der Gasphase vollstandig versintert. Nach Grosenklassierung mittels Differential Mobility Analyzer wurden die Partikeln bei unterschiedlichen Knudsen-Zahlen in einem Impaktor abgeschieden. In der Abb. sind Extremfalle der Bildung von SilberNanopartikelschichten aus der Gasphase gezeigt: (links) hochporose Schicht hergestellt uber Diffusionsabscheidung und (rechts) sehr kompakte Schicht hergestellt uber ballistische Impaktion.