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Diffusion in simple fluids / R. J. Speedy; F. X. Prielmeier; T. Vardag; E. W. Lang; H.-D. Lüdemann
(1989)
Differentiation between Phaeocystis pouchetii (Har.) Lagerheim and Phaeocystis globosa Scherffel
(1987)
The objective of this study is the establishment of a differential scanning calorimetry (DSC) based method for online analysis of the biodegradation of polymers in complex environments. Structural changes during biodegradation, such as an increase in brittleness or crystallinity, can be detected by carefully observing characteristic changes in DSC profiles. Until now, DSC profiles have not been used to draw quantitative conclusions about biodegradation. A new method is presented for quantifying the biodegradation using DSC data, whereby the results were validated using two reference methods.
The proposed method is applied to evaluate the biodegradation of three polymeric biomaterials: polyhydroxybutyrate (PHB), cellulose acetate (CA) and Organosolv lignin. The method is suitable for the precise quantification of the biodegradability of PHB. For CA and lignin, conclusions regarding their biodegradation can be drawn with lower resolutions. The proposed method is also able to quantify the biodegradation of blends or composite materials, which differentiates it from commonly used degradation detection methods.
Die Bereitstellung von nachhaltig erzeugtem Wasserstoff als Energieträger und Rohstoff ist eine wichtige Schlüsseltechnologie sowohl als Ersatz für fossile Energieträger, aber auch als Produkt im Zusammenhang mit Kreislaufprozessen. In der Abwasserbehandlung bestehen verschiedene Möglichkeiten Wasserstoff herzustellen. Mehrere Wege, mögliche Synergien, aber auch deren Nachteile werden vorgestellt.
Kowollik, Silvia ; Schnitzler, Thomas ; Biselli, Manfred ; Krueger, R. ; Zang, Werner ; Peuscher, A. ; Schillberg, S. ; Fischer, R.
In der biopharmazeutischen Industrie werden rekombinante Proteine und monoklonale Antikörper in Zellkulturfermentationen produziert, da nur humane oder tierische Zelllinien über die Fähigkeit der Glykosylierung verfügen. Um hohe Produktausbeuten in ausgezeichneter Qualität zu erzielen, ist eine funktionstüchtige Prozesskontrolle unerlässlich. Hierzu wurde in Kooperation mit der Firma Hitec Zang GmbH die HiSense Präzisionsabgasanalytik entwickelt, die auf Basis der vollautomatischen Ermittlung des Respirationsquotienten (RQ; Verhältnis vonKohlendioxidbildungsrate (CER) zu Sauerstoffaufnahmerate (OTR)) einen Fermentationsprozess nicht-invasiv überwacht. Der RQ kann in Hybridoma- und CHO-Zellen (s. Abb.) in sowohl serumhaltigen als auch serumfreien Medien erfolgreich ermittelt werden. Hier spiegeln die CER und die OTR das Wachstumsverhalten der kultivierten CHO-Zellen wider. Der RQ nimmt dabei Werte zwischen 0,9 und 1,2 an. Dies lässt auf verschiedene Stoffwechselaktivitäten schließen. Da die momentane industrielle Prozesskontrolle auf gemessenen Sauerstoffaufnahmeraten oder entsprechende Offline-Analytiken der Metaboliten basieren, soll durch die vollautomatische RQ-Ermittlung ein neues Verfahren zur Fermentationsüberwachung etabliert werden. Bisher war diese, in bakteriellen Kultivierungen standardisierte Methode, aufgrund der schwierigen CER-Berechnung bei Zellkulturen keine adäquate Alternative.
Three amperometric biosensors have been developed for the detection of L-malic acid, fumaric acid, and L -aspartic acid, all based on the combination of a malate-specific dehydrogenase (MDH, EC 1.1.1.37) and diaphorase (DIA, EC 1.8.1.4). The stepwise expansion of the malate platform with the enzymes fumarate hydratase (FH, EC 4.2.1.2) and aspartate ammonia-lyase (ASPA, EC 4.3.1.1) resulted in multi-enzyme reaction cascades and, thus, augmentation of the substrate spectrum of the sensors. Electrochemical measurements were carried out in presence of the cofactor β-nicotinamide adenine dinucleotide (NAD+) and the redox mediator hexacyanoferrate (III) (HCFIII). The amperometric detection is mediated by oxidation of hexacyanoferrate (II) (HCFII) at an applied potential of + 0.3 V vs. Ag/AgCl. For each biosensor, optimum working conditions were defined by adjustment of cofactor concentrations, buffer pH, and immobilization procedure. Under these improved conditions, amperometric responses were linear up to 3.0 mM for L-malate and fumarate, respectively, with a corresponding sensitivity of 0.7 μA mM−1 (L-malate biosensor) and 0.4 μA mM−1 (fumarate biosensor). The L-aspartate detection system displayed a linear range of 1.0–10.0 mM with a sensitivity of 0.09 μA mM−1. The sensor characteristics suggest that the developed platform provides a promising method for the detection and differentiation of the three substrates.
Cupriavidus necator H16 gains increasing attention in microbial research and biotechnological application due to its diverse metabolic features. Here we present a tightly controlled gene expression system for C. necator including the pBBR1-vector that contains hybrid promoters originating from C. necator native tolC-promoter in combination with a synthetic tetO-operator. The expression of the reporter gene from these plasmids relies on the addition of the exogenous inducer doxycycline (dc). The novel expression system offers a combination of advantageous features as; (i) high and dose-dependent recombinant protein production, (ii) tight control with a high dynamic range (On/Off ratio), which makes it applicable for harmful pathways or for toxic protein production, (iii) comparable cheap inducer (doxycycline, dc), (iv) effective at low inducer concentration, that makes it useful for large scale application, (v) rapid, diffusion controlled induction, and (vi) the inducer does not interfere within the cell metabolism. As applications of the expression system in C. necator H16, the growth ability on glycerol was enhanced by constitutively expressing the E. coli glpk gene-encoding for glycerol kinase. Likewise, we used the system to overcome the expression toxicity of mevalonate pathway in C. necator H16. With this system, the mevalonate-genes were successfully introduced in the host and the recombinant strains could produce about 200 mg/l mevalonate.
An enzyme-based multi-parameter biosensor is developed for monitoring the concentration of formate, d-lactate, and l-lactate in biological samples. The sensor is based on the specific dehydrogenation by an oxidized β-nicotinamide adenine dinucleotide (NAD+)-dependent dehydrogenase (formate dehydrogenase, d-lactic dehydrogenase, and l-lactic dehydrogenase, respectively) in combination with a diaphorase from Clostridium kluyveri (EC 1.8.1.4). The enzymes are immobilized on a platinum working electrode by cross-linking with glutaraldehyde (GA). The principle of the determination scheme in case of l-lactate is as follows: l-lactic dehydrogenase (l-LDH) converts l-lactate into pyruvate by reaction with NAD+. In the presence of hexacyanoferrate(III), the resulting reduced β-nicotinamide adenine dinucleotide (NADH) is then regenerated enzymatically by diaphorase. The electrochemical detection is based on the current generated by oxidation of hexacyanoferrate(II) at an applied potential of +0.3 V vs. an Ag/AgCl reference electrode. The biosensor will be electrochemically characterized in terms of linear working range and sensitivity. Additionally, the successful practical application of the sensor is demonstrated in an extract from maize silage.