Biotoxicity and low temperature are among the challenges of biodegradation as well as microbial transport in in-situ bioremediation applications. This study investigates the effect of low temperature, high and low concentrations of phenol, on biodegradability and the surface properties of Rhodotorula glutinis which may be instrumental to develop control mechanisms in engineering applications. Results showed that Rhodotorula glutinis is able to grow at low phenol (< 0.5 g/L) and glucose (1.0 g/L) concentrations. Microbial growth rates of Rhodotorula glutinis is the highest at 25°C for phenol fed reactors. Temperature and type of carbon source affected the total amount of extracellular polymeric substances (EPS) and the relative amounts of EPS constituents. Phenol fed Rhodotorula glutinis, at 4°C, were found to be hydrophobic whereas Rhodotorula glutinis showed less hydrophobic characteristics at 25°C, regardless of the carbon source. Surface hydrophobicity of Rhodotorula glutinis is related to the distribution of carbohydrate and proteins in the EPS.