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Related Experiment Videos

Ammonia accumulation in acetate-growing yeast.

E Bogonez, A Machado, J Satrústegui

    Biochimica Et Biophysica Acta
    |September 7, 1983
    PubMed
    Summary

    Yeast growing on acetate increase culture pH by losing H+ ions. This alkaline environment drives ammonia accumulation through both ammonium (NH+4) transport and ammonia (NH3) diffusion, impacting yeast physiology.

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    Area of Science:

    • Microbiology
    • Biochemistry
    • Cell Physiology

    Background:

    • Yeast cultures grown on acetate experience a significant pH increase during the stationary phase, shifting from acidic to alkaline conditions (pH 5.8 to 7-8).
    • This pH shift is primarily due to the uptake of acetic acid and the subsequent loss of protons (H+) from the cells.
    • Acetate-grown yeast also exhibit ammonia accumulation, a phenomenon that requires further investigation into its underlying mechanisms.

    Purpose of the Study:

    • To investigate the influence of pH on ammonia transport and accumulation in yeast.
    • To elucidate the mechanisms responsible for ammonia accumulation in alkaline yeast cultures.
    • To characterize the role of pH-dependent transport systems in yeast metabolism.

    Main Methods:

    • Utilized [14C]methylamine as an analogue to study ammonia transport kinetics and accumulation.
    • Measured methylamine uptake and efflux across a range of pH values (acidic to alkaline).
    • Determined internal pH of yeast cells under various culture conditions to understand the driving forces for ammonia accumulation.

    Main Results:

    • Methylamine uptake exhibited a pH-dependent transition from a single-component system (NH+4 permease) at acidic pH (<6.5) to a two-component system (NH+4 permease and NH3 diffusion) at alkaline pH (>7.5).
    • Equilibrium accumulation of methylamine increased with rising external pH, indicating enhanced trapping at higher pH.
    • Methylamine efflux from loaded cells was inversely correlated with external pH, decreasing as pH increased.

    Conclusions:

    • Ammonia accumulation in acetate-grown yeast under alkaline conditions is a result of two combined processes: energy-dependent transport of ammonium (NH+4) and passive diffusion of ammonia (NH3) driven by the pH gradient (delta pH).
    • The study highlights the critical role of pH regulation and membrane transport in managing nitrogenous compounds within yeast cells.
    • Understanding these mechanisms is crucial for optimizing yeast fermentation processes and managing cellular homeostasis.

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