Immobilization of microorganisms on/in insoluble carriers is widely used to stabilize functional activity of microbial cells in industrial biotechnology. We immobilized Rhodococcus ruber, an important hydrocarbon degrader, on biosurfactant-coated sawdust. A biosurfactant produced by R. ruber in the presence of liquid hydrocarbons was found to enhance rhodococcal adhesion to solid surfaces, and thus, it was used as a hydrophobizing agent to improve bacterial attachment to a sawdust carrier. Compared to previously used hydrophobizers (drying oil and n-hexadecane) and emulsifiers (methyl- and carboxymethyl cellulose, poly(vinyl alcohol), and Tween 80), Rhodococcus biosurfactant produced more stable and homogenous coatings on wood surfaces, thus resulting in higher sawdust affinity to hydrocarbons, uniform monolayer distribution of immobilized R. ruber cells (immobilization yield 29-30 mg dry cells/g), and twofold increase in hydrocarbon biooxidation rates compared to free rhodococcal cells. Two physical methods, i.e., high-resolution profilometry and infrared thermography, were applied to examine wood surface characteristics and distribution of immobilized R. ruber cells. Sawdust-immobilized R. ruber can be used as an efficient biocatalyst for hydrocarbon transformation and degradation.