Study of NADH Stability Using Ultraviolet–Visible Spectrophotometric Analysis and Factorial Design

Rover, Laércio; Fernandes, Julio Cesar B.; Neto, Graciliano de Oliveira; Kubota, Lauro T.; Katekawa, Edson; Serrano, Sı́lvia Helena Pires

doi:10.1006/abio.1998.2656

Cited by 121 publications

(86 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In general, monooxygenase enzymes are 1,4-dihydronicotinamde adenine dinucleotide co-factor dependent enzymes; i. e., NADH or NAD(P)H. These co-factors are costly and hydrolytically unstable, and therefore, represent a major barrier for 20 preparative applications utilizing monooxygenases [4,7]. The ribose, pyrophosphate, and adenosine groups (Figure 1) entails the complex structure of NADH, 2b, and NAD(P)H, 3b, thus making chemical synthesis or isolation from biological sources tedious, and creates the high costs of these biocatalytic processes.…”

Section: Fish Jomc Manuscript Revisedmentioning

confidence: 99%

Bioorganometallic chemistry: biocatalytic oxidation reactions with biomimetic NAD+/NADH co-factors and [Cp*Rh(bpy)H]+ for selective organic synthesis

Lutz

Hollmann

Schnyder

et al. 2004

Journal of Organometallic Chemistry

130

102

View full text Add to dashboard Cite

show abstract

Section: Fish Jomc Manuscript Revisedmentioning

confidence: 99%

Bioorganometallic chemistry: biocatalytic oxidation reactions with biomimetic NAD+/NADH co-factors and [Cp*Rh(bpy)H]+ for selective organic synthesis

Lutz

Hollmann

Schnyder

et al. 2004

Journal of Organometallic Chemistry

130

102

View full text Add to dashboard Cite

show abstract

“…Upon incubation at 37 • C for 3 h, an apparent decrease of absorption intensity at 340 nm and a simultaneous increase of absorption at 260 nm were observed ( Figure S7). Such variation was once referred to conversion of NADH to NAD + [31]. However, subsequent 1 H NMR studies indicated that a new compound besides NAD + was formed.…”

Section: Proposed Mechanism Of Nadh Degradationmentioning

confidence: 99%

Ionic Liquids as Bifunctional Cosolvents Enhanced CO2 Conversion Catalysed by NADH-Dependent Formate Dehydrogenase

Zhang

Luo

et al. 2018

Catalysts

View full text Add to dashboard Cite

Efficient CO 2 conversion by formate dehydrogenase is limited by the low CO 2 concentrations that can be reached in traditional buffers. The use of ionic liquids was proposed as a manner to increase CO 2 concentration in the reaction system. It has been found, however, that the required cofactor (NADH) heavily degraded during the enzymatic reaction and that acidity was the main reason. Acidity, indeed, resulted in reduction of the conversion of CO 2 into formic acid and contributed to overestimate the amount of formic acid produced when the progression of the reaction was followed by a decrease in NADH absorbance (method N). Stability of NADH and the mechanism of NADH degradation was investigated by UV, NMR and by DFT calculations. It was found that by selecting neutral-basic ionic liquids and by adjusting the concentration of the ionic liquid in the buffer, the concentration of NADH can be maintained in the reaction system with little loss. Conversion of CO 2 to methanol in BmimBF 4 (67.1%) was more than twice as compared with the conversion attained by the enzymatic reaction in phosphate buffer (24.3%).

show abstract

“…This behavior is presumably due to ongoing chemistry leading to variable conversion in the atomizer volume between different oxidation states of NADH, only one of which is fluorescent. While NADH-fluorescence is widely used to study enzymatic reactions (Lawkowicz et al, 1992), it is known to be highly unstable in solution (Rover, 1998) and the oxidized form (NAD+) is not fluorescent (Rost, 1992). So while NADH is an appealing potential calibrant for the WIBS, since it is a common metabolic molecule that exhibits fluorescence in the FL2 detector following both excitation wavelengths, stability issues prevented us from fully assessing it as we have done for quinine and tryptophan.…”

Section: Failed Calibrants: Nadh and Naphthalenementioning

confidence: 99%

Fluorescence calibration method for single-particle aerosol fluorescence instruments

et al. 2017

View full text Add to dashboard Cite

Abstract. Real-time, single-particle fluorescence instruments used to detect atmospheric bioaerosol particles are increasingly common, yet no standard fluorescence calibration method exists for this technique. This gap limits the utility of these instruments as quantitative tools and complicates comparisons between different measurement campaigns. To address this need, we have developed a method to produce size-selected particles with a known mass of fluorophore, which we use to calibrate the fluorescence detection of a Wideband Integrated Bioaerosol Sensor (WIBS-4A). We use mixed tryptophan-ammonium sulfate particles to calibrate one detector (FL1; excitation = 280 nm, emission = 310-400 nm) and pure quinine particles to calibrate the other (FL2; excitation = 280 nm, emission = 420-650 nm). The relationship between fluorescence and mass for the mixed tryptophan-ammonium sulfate particles is linear, while that for the pure quinine particles is nonlinear, likely indicating that not all of the quinine mass contributes to the observed fluorescence. Nonetheless, both materials produce a repeatable response between observed fluorescence and particle mass. This procedure allows users to set the detector gains to achieve a known absolute response, calculate the limits of detection for a given instrument, improve the repeatability of the instrumental setup, and facilitate intercomparisons between different instruments. We recommend calibration of single-particle fluorescence instruments using these methods.

show abstract

Study of NADH Stability Using Ultraviolet–Visible Spectrophotometric Analysis and Factorial Design

Cited by 121 publications

References 34 publications

Bioorganometallic chemistry: biocatalytic oxidation reactions with biomimetic NAD+/NADH co-factors and [Cp*Rh(bpy)H]+ for selective organic synthesis

Bioorganometallic chemistry: biocatalytic oxidation reactions with biomimetic NAD+/NADH co-factors and [Cp*Rh(bpy)H]+ for selective organic synthesis

Ionic Liquids as Bifunctional Cosolvents Enhanced CO2 Conversion Catalysed by NADH-Dependent Formate Dehydrogenase

Fluorescence calibration method for single-particle aerosol fluorescence instruments

Contact Info

Product

Resources

About