Pulsed Electric Field Inactivation of Acetobacter aceti: Mechanisms and Efficacy Elucidation

Abstract

The spoilage bacterium Acetobacter aceti poses a major threat to wine quality by causing acetification, driving the need for effective control methods. This study investigated the inactivation of A. aceti using pulsed electric field (PEF) and elucidated the multi-target mechanisms. The results demonstrated that PEF efficacy was highly dependent on the electric field intensity. PEF treatment at 30 kV/cm achieved a >3-log reduction in viable cell counts, with a Weibull model analysis indicating a critical inactivation threshold of 21.64 kV/cm. Mechanistic investigations revealed that PEF induced irreversible damage to the cell membrane, evidenced by morphological rupture (SEM) and a 4-fold increased permeability (flow cytometry), which led to a massive leakage of intracellular contents. Critically, this physical damage was correlated with profound physiological disruption, including the inactivation of key spoilage enzymes alcohol dehydrogenase (ADH, 80.0% relative activity loss) and aldehyde dehydrogenase (ALDH, 93.1% relative activity loss). Furthermore, PEF induced severe oxidative stress (4.25-fold increase in ROS and 3.30-fold increase in MDA) and a collapse in energy metabolism. Collectively, these findings reveal a synergistic inactivation mechanism, which establishes a strong theoretical foundation for the potential development of PEF as a non-thermal strategy to control acetic spoilage in winemaking.