The overall objective of this work was to investigate the modifications that occur in Bacillus subtilis lipid membrane during induced oxidative stress caused by reactive oxygen species (ROS), via an electrophysiological approach. Further, based on the results, we have developed and demonstrated a novel strategy to enhance specific enzyme production. Electrical parameters such as phase angle (θ), impedance (Z), capacitance (C) and breakdown voltage of reconstituted bilayer lipid membrane (BLM) composed of lipids extracted from non-stressed, mildly stressed (2.5 mM H2O2) and strongly stressed (2.5 mM H2O2 with 100 μM FeSO4) B. subtilis were compared. Strongly stressed BLM showed lower values of θ (10°), Z (0.4 Mohm), and breakdown voltage (100 mV) in comparison with those observed for non-stressed BLM, i.e., 30°, 0.5 Mohm and 250 mV, respectively. The capacitance of strongly stressed BLM, however, was higher (2.28 nF) compared to that of the non-stressed BLM (0.4 nF). These results suggest that under strongly stressed conditions, the lipids were loosely packed that resulted in a more permeable BLM. The higher permeability seems to result, unexpectedly, from a higher unsaturated fatty acid (UFA) synthesis and membrane incorporation (UFA fraction increased by 227%), and expectedly, from increased lipid peroxidation (increased by nearly 200%) in the BLM. A strategy that is based on increased membrane permeability due to induced ROS, enhanced specific amylase and protease production under oxidative stress by 62% and 137%, respectively. © 2009 Elsevier Ltd. All rights reserved.