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Related Concept Videos

Balancing Redox Equations02:58

Balancing Redox Equations

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Electrochemistry is the science involved in the interconversion of electrical and chemical reactions. Such reactions are called reduction-oxidation, or redox reactions. These important reactions are defined by changes in oxidation states for one or more reactant elements and include a subset of reactions involving the transfer of electrons between reactant species. Electrochemistry as a field has evolved to yield sufficient insights on the fundamental principles of redox chemistry and multiple...
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Redox Reactions01:24

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Oxidation-reduction or redox reactions involve the transfer of electrons from one molecule or atom to another. When an atom gains an electron, another atom must lose an electron, meaning oxidation and reduction must occur together. Since the redox occurs in pairs, the atom that gets oxidized is also called the reducing agent or reductant, and the atom that is reduced is also called the oxidizing agent or oxidant. A straightforward way to remember the definitions of oxidation and reduction is...
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Redox Reactions01:27

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Redox reactions are vital biochemical processes that underpin energy metabolism in cells. These reactions involve the transfer of electrons between molecules, occurring in tandem as oxidation and reduction. Oxidation refers to the loss of electrons, while reduction denotes their gain. This coupling ensures the seamless flow of electrons through metabolic pathways. For example, in bacterial metabolism, glucose undergoes oxidation to carbon dioxide, while oxygen is simultaneously reduced to...
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Cis-regulatory Sequences02:02

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Cis-regulatory sequences are short fragments of non-coding DNA that are present on the same chromosomes as the genes that they regulate. These fragments serve as binding sites for transcriptional regulators, proteins that are responsible for controlling gene transcription and differential gene expression across cell types in eukaryotes. Cis-regulatory sequences can be close to the gene of interest or thousands of bases away in the DNA sequence; however, those sequences that are further away are...
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Cis-regulatory Sequences

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Electron Carriers01:24

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Electron carriers can be thought of as electron shuttles. These compounds can easily accept electrons (i.e., be reduced) or lose them (i.e., be oxidized). They play an essential role in energy production because cellular respiration is contingent on the flow of electrons.
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Updated: Jan 30, 2026

Ratiometric Biosensors that Measure Mitochondrial Redox State and ATP in Living Yeast Cells
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A Regulatory NADH/NAD+ Redox Biosensor for Bacteria.

Yang Liu, Robert Landick, Srivatsan Raman

    ACS Synthetic Biology
    |January 12, 2019
    PubMed
    Summary

    Researchers developed a novel biosensor to easily measure the NADH/NAD+ ratio, a key indicator of cellular health. This tool enables high-throughput screening for metabolic engineering and understanding cellular redox states.

    Keywords:
    NADH/NAD+bacterial physiologyhigh-throughput screeningredox biosensorrespiratory chain

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    Area of Science:

    • Biochemistry
    • Metabolic Engineering
    • Synthetic Biology

    Background:

    • The ratio of NADH to NAD+ is a critical indicator of cellular metabolic state, influencing numerous biochemical reactions.
    • Existing methods for measuring the NADH/NAD+ ratio are often labor-intensive, inaccurate, and unsuitable for high-throughput applications.

    Purpose of the Study:

    • To develop a genetically encoded, ratiometric biosensor for facile and high-throughput measurement of the NADH/NAD+ ratio.
    • To engineer and characterize a novel biosensor based on the redox-responsive bacterial transcription factor Rex.
    • To apply the biosensor to investigate cellular redox states under different metabolic conditions and genetic perturbations.

    Main Methods:

    • Engineered a Rex-regulated promoter in E. coli to optimize biosensor performance by tuning transcription factor affinity and operator site.
    • Utilized the biosensor-reporter system to assess the impact of respiratory chain enzyme deletions on the NADH/NAD+ ratio during aerobic respiration.
    • Investigated the influence of various carbon sources (acetate vs. glucose) on the cellular NADH/NAD+ ratio.
    • Demonstrated high-throughput screening capability by enriching for high-NADH mutants using a pooled screen approach.

    Main Results:

    • The engineered Rex biosensor provides improved characteristics for NADH/NAD+ ratio measurement.
    • Mutations in respiratory chain enzymes led to significant increases (over 3-fold) in the NADH/NAD+ ratio, with a double NADH dehydrogenase mutant showing a 6-fold elevation.
    • E. coli grown on acetate exhibited a higher NADH/NAD+ ratio compared to growth on glucose.
    • Successfully enriched rare high-NADH mutants (1 in 10,000) using biosensor-guided pooled screening, demonstrating the system's utility for high-throughput screening.

    Conclusions:

    • The developed Rex biosensor-reporter offers a noninvasive, facile, and high-throughput method for measuring cellular redox states.
    • This tool facilitates a deeper understanding of redox metabolism and enables engineering of cellular redox states.
    • The biosensor is a powerful platform for screening and identifying cellular variants with altered metabolic profiles.