Microalgae have a high biotechnological potential as a source of biofuels (biodiesel, biohydrogen) and other high-added value products (e.g., pharmaceuticals, proteins, pigments). However, for microalgae cultivation to be economically competitive with other fuel sources, it is necessary to apply the concept of biorefinery. This seems to be the most ambitious strategy to achieve viability. Therefore, the objectives of this study were to isolate and identify the main microalgae line used to produce biofuels at Federal University of Parana, Brazil, using the rDNA sequence and micromorphological analysis, and to evaluate the potential of this lineage in the production of hydrogen and co-products with biological activity. For the purification of the lineage (LGMM0001), an aliquot was seeded into solid CHU culture medium and an isolated colony was selected. The genomic DNA was purified using a commercial kit (Macherey-Nagel, Düren, Germany) for molecular identification, the ITS region (ITS1, 5.8S and ITS2) (Internal Transcribed Spacer) was amplified and sequenced using primers LS266 and V9G. Morphological characterization was performed as described by Hemschemeier et al. . Finally, for biological activity research, secondary metabolites were extracted by fractionation and evaluated against bacteria of clinical interest. Through microscopic analysis, general characteristics shared by the genus Tetradesmus were observed. The plasticity of the morphological characteristics of this genus reinforces the need for further studies to classify correctly the species in this group, using DNA sequencing. ITS sequence analysis of LGMM0001 showed 100% homology with sequences from the Tetradesmus obliquus species, so, the lineage was classified as belonging to this species. The evaluated microalgae strain was able to produce hydrogen, showing positive results for gas formation. Biological activity was observed with the extract obtained from the residual culture carried out with alternative medium used in the photobioreactors (PBR), against the Staphylococcus aureus pathogenic lineage. In conclusion, the microalgae strain used in this work was identified as Tetradesmus obliquus (= Acutodesmus obliquus), and was able to produce a compound with economic potential in association with the existing biofuel production process.