Two-Step Macromolecule Separation Process with Acid Pretreatment and High-Shear-Assisted Extraction for Microalgae-Based Biorefinery

A simple two-stage extraction and recovery method for macromolecules from microalgae biomass, termed CASS (concentrating the microalgae solution, acid pretreatment, high-shear-assisted lipid extraction, and separation), was developed. This method effectively processed the wet biomass of Chlorella sp. ABC-001 at a moderately low biomass concentration (50 g/L). The optimal conditions were acid pretreatment with 5 wt.% H₂SO₄ at 100 °C for 1 h, followed by high-shear extraction using hexane at 3000 rpm for 30 min. The acid pretreatment hydrolyzed carbohydrates and phospholipids, disrupting the cell wall and membrane, while high-shear mixing enhanced mass transfer rates between solvents and lipids, overcoming the hydraulic barrier at the cell surface. Within 10 min after completing the process, the extraction mixture achieved natural phase separation into water, solvent, and biomass residue layers, each enriched with carbohydrates, lipids, and proteins, respectively. The CASS process demonstrated high esterifiable lipid yields (91%), along with substantial recovery of glucose (90%) and proteins (100%). The stable phase separation prevented emulsion formation, simplifying downstream processing. This study presents the results on cell disruption, optimal acid treatment concentration, and high-shear mixing to achieve macromolecule separation, expanding the lipid-centric microalgal process to a comprehensive biorefinery concept.