Electrochemical probing of in vivo 5-hydroxymethyl furfural reduction in Saccharomyces cerevisiae

Analytical Chemistry
Natalie KosteshaJenny Emneus

Abstract

In this work, mediated amperometry was used to evaluate whether differences in intracellular nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) level could be observed between a genetically modified Saccharomyces cerevisiae strain, engineered for NADPH dependent 5-hydroxymethyl-2-furaldehyde (HMF) reduction, and its control strain. Cells overexpressing the alcohol dehydrogenase 6 gene (ADH6 strain) and cells carrying the corresponding control plasmid (control strain) were each immobilized on Au-microelectrodes. The real-time dynamics of NAD(P)H availability in the two strains, preincubated with HMF, was probed using the menadione-ferricyanide double mediator system. A lower intracellular NADPH level as the consequence of more effective HMF reduction was observed for the ADH6 strain both with and without added glucose, which increases the overall cellular NADPH level. The mediated amperometric signal during real-time monitoring of the concentration dependent HMF reduction in living cells could be translated into the cellular enzyme kinetic parameters: K(M,cell)(app), V(MAX), k(cat,cell), and k(cat,cell)/K)M,cell)(app). The results indicated that the overexpression of the ADH6 gene gave a 68% decrease in K(M,cell)(app) and 4...Continue Reading

References

Sep 1, 1977·Archives of Biochemistry and Biophysics·H Y TingM K Jacobson
Jan 30, 1987·Clinica Chimica Acta; International Journal of Clinical Chemistry·J Racek, J Musil
Jan 28, 1999·Biotechnology and Bioengineering·E PalmqvistB Hahn-Hägerdal
Aug 5, 2000·Applied Microbiology and Biotechnology·M J TaherzadehG Lidén
Aug 23, 2002·Functional & Integrative Genomics·Timothy R Hughes
Apr 30, 2005·Trends in Pharmacological Sciences·Willem H Mager, Joris Winderickx
Oct 10, 2006·Applied Microbiology and Biotechnology·Z Lewis Liu
Mar 12, 2008·Applied Microbiology and Biotechnology·João R M AlmeidaMarie-F Gorwa-Grauslund

❮ Previous
Next ❯

Citations

Jan 19, 2013·Analytical and Bioanalytical Chemistry·Roland LudwigLo Gorton
Feb 28, 2013·Analytical and Bioanalytical Chemistry·Mehdi RahimiSusan R Mikkelsen
Aug 27, 2014·FEMS Yeast Research·Adebola AdeniranKeith E J Tyo

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.