Selecting the optimal microalgal strain for carbon capture and biomass production is crucial for ensuring the commercial viability of microalgae‐based biorefinery processes. This study aimed to evaluate the impact of varying bicarbonate concentrations on the growth rates, inorganic carbon (IC) utilization, and biochemical composition of three freshwater and two marine microalgal species. Parachlorella kessleri, Vischeria cf. stellata, and Porphyridium purpureum achieved the highest carbon removal efficiency (>85%) and biomass production at 6 g L⁻¹ sodium bicarbonate (NaHCO₃), while Phaeodactylum tricornutum showed optimal performance at 1 g L⁻¹ NaHCO₃. The growth and carbon removal rate of Scenedesmus quadricauda increased with increasing NaHCO₃ concentrations, although its highest carbon removal efficiency (∼70%) was lower than the other species. Varying NaHCO₃ levels significantly impacted the biochemical composition of P. kessleri, S. quadricauda, and P. purpureum but did not affect the composition of the remaining species. The fatty acid profiles of the microalgae were dominated by C16 and C18 fatty acids, with P. purpureum and P. tricornutum yielding relatively high polyunsaturated fatty acid content ranging between 14% and 30%. Furthermore, bicarbonate concentration had a species‐specific effect on the fatty acid and chlorophyll‐a content. This study demonstrates the potential of bicarbonate as an effective IC source for microalgal cultivation, highlighting its ability to select microalgal species for various applications based on their carbon capture efficiency and biochemical composition.