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A flow system for substrate recycling of NAD+/NADH was set up with an enzyme reactor containing coimmobilized glycerol dehydrogenase (GDH) and diaphorase. The product from the diaphorase catalysis, hexacyanoferrate(II), aws detected amperometrically at a glassy carbon electrode. The amplification factor was 150 for a reactor volume of 100 μ l at a flow-rate of 0.5 ml/min. With a stopped flow of four minutes, the signal increased another 88 times, resulting in a signal amplification of 13 300 times. Equations are derived for the amplification factor and used for a discussion of the optimization of amplification systems. The Km for GDH with glycerol as a substrate was found to be 5 × 10−3 M at pH 8.0. GDH from Cellulomonas sp. was purified on a gel filtration column and the purified enzyme showed a specificity toward NAD+, compared to NADP+, that was higher than 99.9%. Due to the NAD+ specificity of the purified GDH, the enzyme amplification system reported here could be used in detection systems for enzyme immunoassays when using alkaline phosphatase as a label and NADP+ as a substrate. The stability of immobilized GDH and diaphorase is several orders of magnitude better than that of alcohol dehydrogenase, which is the enzyme commonly used for NAD+-specific detection in these applications. 相似文献
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Carl‐Johan Wallentin Torbjörn Wixe Ola F. Wendt Dr. Karl‐Erik Bergquist Dr. Kenneth Wärnmark Dr. 《Chemistry (Weinheim an der Bergstrasse, Germany)》2010,16(13):3994-4002
A pair of molecular tweezers (syn‐ 4 ) that consists of quinoline and pyrazine units fused to a bicyclic framework is presented. The tweezers were synthesised both as a racemic mixture (rac‐ 4 ) and an enantiomerically pure form ((R,R,R,R)‐ 4 ) starting from either racemic or enantiomerically pure bicyclo[3.3.1]nonane‐2,6‐dione ( 3 ). Homochiral dimers were observed in the solid state for rac‐ 4 . The self‐association of both rac‐ 4 and (R,R,R,R)‐ 4 was studied in solution. A weak self‐association constant in CDCl3 was estimated by 1H NMR spectroscopic dilution titration experiments in both cases, following several proton resonances. For this purpose, a general normalisation model for the accurate determination of association constants from multiple datasets was developed. In contrast to the solid state, no diastereomeric discrimination was observed for rac‐ 4 in solution. 相似文献
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Wallentin CJ Nguyen JD Finkbeiner P Stephenson CR 《Journal of the American Chemical Society》2012,134(21):8875-8884
Herein, the development of visible light-mediated atom transfer radical addition (ATRA) of haloalkanes onto alkenes and alkynes using the reductive and oxidative quenching of [Ir{dF(CF(3))ppy}(2)(dtbbpy)]PF(6) and [Ru(bpy)(3)]Cl(2) is presented. Initial investigations indicated that the oxidative quenching of photocatalysts could effectively be utilized for ATRA, and since that report, the protocol has been expanded by broadening the scope of the reaction in terms of the photocatalysts, substrates, and solvents. In addition, further modifications of the reaction conditions allowed for the efficient ATRA of perfluoroalkyl iodides onto alkenes and alkynes utilizing the reductive quenching cycle of [Ru(bpy)(3)]Cl(2) with sodium ascorbate as the sacrificial electron donor. These results signify the complementary nature of the oxidative and reductive quenching pathways of photocatalysts and the ability to predictably direct reaction outcome through modification of the reaction conditions. 相似文献
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Dr. Anna Hedström Dr. Zakieh Izakian Irma Vreto Dr. Carl‐Johan Wallentin Prof. Dr. Per‐Ola Norrby 《Chemistry (Weinheim an der Bergstrasse, Germany)》2015,21(15):5946-5953
The radical nature of iron‐catalyzed cross‐coupling between Grignard reagents and alkyl halides has been studied by using a combination of competitive kinetic experiments and DFT calculations. In contrast to the corresponding coupling with aryl halides, which commences through a classical two‐electron oxidative addition/reductive elimination sequence, the presented data suggest that alkyl halides react through an atom‐transfer‐initiated radical pathway. Furthermore, a general iodine‐based quenching methodology was developed to enable the determination of highly accurate concentrations of Grignard reagents, a capability that facilitates and increases the information output of kinetic investigations based on these substrates. 相似文献
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The technique of ferromagnetic resonance at 23 GHz has been used to determine the first three anisotropy constants of pure Ni down to 4.2K. A temperature and orientation dependent linewidth has also been observed. 相似文献