In this work, proteomic analysis was used to identify the up-regulated key proteins of Pseudomonas aeruginosa (P6), a bacteria used in petroleum degradation, responsible for its high efficiency in degrading crude oil. Seventeen proteins were identified as up-regulated proteins by proteomic analysis and classified by bioinformatics analysis. The results indicated that most of the up-regulated proteins were responsible for P. aeruginosa (P6) survival under harsh environmental conditions and utilization crude oil as carbon source in a better way. The physiological processes, chemotaxis to carbon sources, terminal oxidation of carbons, carbon source uptake and nutrients transport, were associated with the up-regulated proteins in the study. The findings revealed the most influential proteins and set a clear direction for future research.