Cellular NADH and NADPH Conformation as a Real-Time Fluorescence-Based Metabolic Indicator under Pressurized...

Area of Science:

• Biophysics
• Cellular metabolism
• Spectroscopy

Background:

• Cellular metabolism and antioxidant defense are crucial for organism survival.
• Understanding how external factors like pressure influence these processes is vital.
• Autofluorescence spectroscopy offers a non-invasive method to probe cellular states.

Purpose of the Study:

• To investigate the real-time metabolic responses of Saccharomyces cerevisiae under high-pressure conditions using autofluorescence spectroscopy.
• To assess the role of pressure in cellular energy metabolism and antioxidant defense.
• To explore the potential of spectral phasor analysis for monitoring metabolic pathways under pressure.

Main Methods:

• UV-excited autofluorescence spectroscopy was employed on Saccharomyces cerevisiae.
• Spectral phasor analysis was used to interpret autofluorescence data.
• Metabolic responses were monitored under varying pressures (ambient, 12 MPa, 30 MPa, 32 MPa) and in the presence of inhibitors like cyanide and ethanol.

Main Results:

• Pressure itself induces changes in cellular redox state and reactive oxygen species (ROS) production.
• The autofluorescence response to cyanide at 32 MPa showed similarities to ambient conditions but with pressure-induced effects.
• Pressurization altered the mechanism of ethanol action, indicated by a shift from two-component to non-two-component spectral behavior.

Conclusions:

• Autofluorescence spectroscopy, particularly spectral phasor analysis, is a valuable tool for real-time metabolic monitoring under pressurized conditions.
• High pressure influences cellular metabolism, redox state, and ROS production in yeast.
• This approach opens new avenues for investigating piezophysiology and cellular responses in agriculture and food technologies.