The synergistic effect of temperature (25–65°C) and total specific energy input (0.55–1.11kWhkgDW−1) by pulsed electric field (PEF) on the release of intracellular components from the microalgae Chlorella vulgaris was studied. The combination of PEF with temperatures from 25 to 55°C resulted in a conductivity increase of 75% as a result of cell membrane permeabilization. In this range of temperatures, 25–39% carbohydrates and 3–5% proteins release occurred and only for carbohydrate release a synergistic effect was observed at 55°C. Above 55°C spontaneous cell lysis occurred without PEF. Combined PEF–temperature treatment does not sufficiently disintegrate the algal cells to release both carbohydrates and proteins at yields comparable to the benchmark bead milling (40–45% protein, 48–58% carbohydrates).