synopsisTimedependent, apparent heat capacities of glucose, poly(viny1 chloride), polystyrene, selenium, poly(methy1 methacrylate), and poly(2,6-diiethyl-1,4-phenylene ether) in the glass transition region were determined by differential thermal analysis. The thermal history was set by linear cooling a t rates between 0.007 and 16OoC/min. Linear heating for analysis was carried out at rates between 0.3 and 6OO0C/min. Average activation energies of 52,81,90,54,77, and 108 kcal/mole, respectively, were evaluated by using the hole theory of glasses previously developed. Within experimental limitations all data could be described quantitatively by the theoretical expressions using only one parameter, the number of frozen-in holes, to describe the thermal history. Experimental and theoretical limitations are discussed.

The purpose of this work was to see if the alkylpyrroleviologen redox polymer technology previously developed for a reagentless nitrate biosensor based on nitrate reductase (NaR) from Escherichia coli (Cosnier, S.; Innocent, C.; Jouanneau, Y. Anal. Chem. 1994, 66, 3198-3201) could be applied to the isozyme from Aspergillus niger. In particular, the enzyme viability after immobilization was of great interest, as Cosnier et al. reported a residual activity of only 0.33% of the amount initially applied. The present work showed that A. niger NaR lost 99.2% of soluble activity on vacuum-drying in the presence of 2.5 nM N-methyl-N'-(12-[pyrrol-1-yl]dodecyl)-4,4'-bipyridinium ditetrafluoroborate monomer (C12V2+) and that most of this loss was due to monomer inhibition (91%). The loss due to dehydration was only 8%. In the biosensor configuration, the enzyme gave a residual activity of 0.18% of the amount originally applied and a specific response of 1.7 mA M-1 cm-2, but all activity was lost after 4 d storage at 4 degrees C in phosphate buffer. It was concluded that for practical biosensors and bioreactors, modification of the redox polymer format was needed, for example by covalent immobilization, to effect higher loading of viable NaR and improved enzyme stability.

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