Immobilization of heavy metals by microbially induced carbonate precipitation using hydrocarbon-degrading ureolytic bacteria

Crude oil contamination introduces multiple threats to human health and the environment, most of which are from toxic heavy metals. Heavy metals cause significant threats because of their persistence, toxicity, and bio-accumulation. Biomineralization, performed through many microbial processes, can lead to the immobilization of heavy metals in formed minerals. The potential of the microbially carbonate-induced precipitation (MICP) in removal by biomineralization of several heavy metals was investigated. A collection of diverse 11 bacterial strains exhibited ureolytic activity and tolerance to heavy metals when growing in Luria-Bertani (LB) and urea medium. Determination of the minimum inhibitory concentrations (MIC) revealed that heavy metal toxicity was arranged as Cd > Ni > Cr > Cu > Zn. Three hydrocarbon-degrading bacterial strains (two of Pseudomonas aeruginosa and one of Providencia rettgeri) exhibited the highest tolerance (MIC > 5 mM) to Cu, Cr, Zn, and Ni, whereas Cd exerted significantly higher toxicity with MIC <1 mM. At all MICP conditions, different proportions of calcium carbonate (calcite) and calcium phosphate (brushite) were formed. Pseudomonas aeruginosa strains (QZ5 and QZ9) exhibited the highest removal efficiency of Cr (100%), whereas Providencia rettgeri strain (QZ2) showed 100% removal of Zn. Heavy metal complexes were found as well. Cd removal was evidenced by the formation of cadmium phosphate induced by Providencia rettgeri bacterial activity. Our study confirmed that hydrocarbon-degrading ureolytic bacteria not only can tolerate heavy metal toxicity but also have the capability to co-precipitate heavy metals. These findings indicate an effective and novel biological approach to bioremediate petroleum hydrocarbons and immobilize multiple heavy metals with mineral formation. This is of high importance for ecological restoration via stabilization of soil and alleviation of heavy metal toxicity.