Staphylococcus aureus is an opportunistic pathogen known to cause an array of acute and chronic infections. The high pathogenicity of this bacterium is thought to be due to its remarkable capacity to rapidly adapt to changes in environmental conditions. Bacterial response mechanisms for survival in changing environments involve genomic and proteomic, metabolomic processes. These responses and adjustments in bacterial systems lead to heterogeneity within the bacterial population, maximising chances of survival. This study used a range of combined stressors including (pH6-8), temperature (35-39 °C) and osmotic stress (0-5% NaCl added), which represent conditions to mimic those on the skin or in a wound site. Experiments were designed by software called MODDE to examine growth under 10 different sets of conditions with variable factors for pH, temperature and osmotic stress. Culture grown under optimal conditions at 37°C, pH7, and no added NaCl were designated the reference control samples and those grown with an additional 2.5% NaCl represented the centroid samples with mid-point values of all variable parameters. The cultures were grown to the mid-exponential phase of growth when they were harvested and extracted to examine the metabolomic and proteomic responses. The extracted metabolites were analysed using Gas Chromatography- Flame Ionization or Mass Spectrometry (GC-FID or GC-MS), and the proteomes were analysed by SDS-PAGE and LC-MS/MS. It was found that each set of environmental conditions generated characteristic metabolite profiles where each of the replicates were tightly clustered and well resolved in multivariate analyses (discriminant function and PCA, P< 0.05). The protein profiles also exhibited characteristic profile responses for each treatment set, indicating that S. aureus had a unique adaptation to each treatment. The data provided strong evidence supporting the hypothesis that specific changes in metabolic homeostasis and protein composition were critical to the adaptive processes required for survival under changing environmental conditions.