EtBE is a fuel oxygenate, made out of (bio)ethanol replacing MtBE. Biodegradation of EtBE can reduce the risk after accidental release in the environment. The oxygen produced by Scenedesmus and Chlorella was used by microorganisms to degrade EtBE using a P450 monooxygenase cytochrome. Metabolites formed during the micro-oxic EtBE degradation were tert-butyl alcohol (TBA), ethanol and CO2 determined using 13C nuclear magnetic resonance spectroscopy (NMR) and gas chromatography. Stable isotope probing (SIP) of the 13C and 12C enriched EtBE fractions showed no significant difference between phylotypes, including Halomonadaceae, Shewanellaceae, Rhodocyclaceae, Oxalobacteraceae, Comamonadaceae, Sphingomonadaceae, Hyphomicrobiaceae, Candidatus Moranbacteria, Omnitrophica, Anaerolineaceae, Nocardiaceae, and Blastocatellaceae. This study is the first study describing micro-oxic degradation of EtBE by an algal-bacterial culture.