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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.
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.
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.
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.