The high-molecular weight polycyclic aromatic hydrocarbons (PAHs) pyrene and typical long chain alkane n-hexadecane are both difficult to degrade. In this study, n-hexadecane and pyrene degrading strain Rhodococcus sp. T1 was isolated from oil contaminated soil. Strain T1 could remove 90.81% n-hexadecane (2 vol%) and 42.79% pyrene (200 mg.L-1) as a single carbon within 5 days, respectively. Comparatively, the degradation of pyrene increased to 60.63%, but the degradation of n-hexadecane decreased to 87.55% when these compounds were mixed. Additionally, identification and analysis of degradation metabolites of Rhodococcus sp. T1 in the above experiments showed that there were significant changes in alanine, methylamine, citric acid and heptadecanoic add between sole and dual substrate degradation. The optimal conditions for degradation were then determined based on analysis of the pH, salinity, additional nutrient sources and liquid surface activity. Under the optimal conditions of pH 7.0, 35 degrees C, 0.5% NaCl, 5 mg.L-1 of yeast extract and 90 mg.L-1 of surfactant, the degradation increased in single or dual carbon sources. To our knowledge, this is the first study to discuss metabolite changes in Rhodococcus sp. T1 using sole substrate and dual substrate to enhance the long-chain alkanes and PAHs degradation potential. (C) 2018 The Chemical Industry and Engineering Society of China, and Chemical Industry Press. All rights reserved.