Rhodococcus erythropolis as a flotation collector for hematite was evaluated in the study. The surface morphology and cell wall composition of R. erythropolis was analyzed. Zeta potentials for four pure minerals from hematite ores were measured before and after adsorption by R. erythropolis. Pure mineral flotation tests and mixed mineral separation tests were performed to describe adsorption characteristics and mechanisms. A rod-shaped bacterium was detected with CH2, CH3 and COO- groups on its cell wall that imparted hydrophobicity and negative charges. The ability of R. erythropolis to collect hematite was stronger than its ability to collect quartz, kaolinite and apatite. For a pulp pH of 6 and a cell concentration of 75 mg/L, recovery rate of hematite was 89.67%. The recovery differences between hematite and quartz, kaolinite and apatite were 66.43%, 60.36% and 54.30%, respectively. These data indicated that electrical properties of hematite surface were more suitable for adsorption of R. erythropolis than other three minerals. The adsorbed hematite particles appeared as large agglomerates after interaction with R. erythropolis. The quartz, kaolinite and apatite particles, however, were in the form of dispersed particles or small agglomerates. The chemical adsorption of hematite on bacterial cell wall resulted in agglomeration. The effects of flocculation and flotation of R. etythropolis on micro-fine hematite particles were characterized for the first time. The results showed that R. etythropolis can act as a flotation collector for hematite from hematite ores. Crown Copyright (C) 2013 Published by Elsevier Ltd. All rights reserved.