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Microalgae accumulate abundant lipids and are a promising source for biodiesel. However, carbohydrates account for 40% of microalgal biomass, an important consideration when using them for the economically feasible production of biodiesel. In this study, different acid hydrolysis and post-treatment processing of Chlorella sp. ABC-001 was performed, and the effect of these different hydrolysates on bioethanol yield by Saccharomyces cerevisiae KL17 was evaluated. For hydrolysis using H2SO4, the neutralization using Ca(OH)2 led to a higher yield (0.43 g ethanol/g sugars) than NaOH (0.27 g ethanol/g sugars). Application of electrodialysis to the H2SO4 + NaOH hydrolysate increased the yield to 0.35 g ethanol/g sugars, and K+ supplementation further enhanced the yield to 0.41 g ethanol/g sugars. Hydrolysis using HNO3 led to the generation of reactive species. Neutralization using only NaOH yielded 0.02 g ethanol/g sugars, and electrodialysis provided only a slight enhancement (0.06 g ethanol/g sugars). However, lowering the levels of reactive species further increased the yield to 0.25 g ethanol/g sugars, and K+ supplementation increased the yield to 0.35 g ethanol/g sugars. Overall, hydrolysis using H2SO4 + Ca(OH)2 provided the highest ethanol yield, and the yield was almost same as from conventional medium. This research emphasizes the importance of post-treatment processing that is modified for the species or strains used for bioethanol fermentation. [ABSTRACT FROM AUTHOR]

• Hydrolyzed H. tuberosus replaced fructose in heterotrophic Ettlia sp. cultivation. • Hydrolysates of algal residue replaced yeast extract as N source. • Sequential hydrolysis was developed to utilize C and N sources simultaneously. • Lipid productivity was enhanced with sequential hydrolysates medium. Heterotrophic cultivation of microalgae results in substantial productivity but requires significant nutrient input. Alternative sources of major nutrients such as C and N should be carefully selected to make heterotrophic production of microalgal biomass feasible. In this study, Helianthus tuberosus and lipid-extracted algae (LEA) were hydrolyzed with sulfuric acid to grow Ettlia sp. YC001. Hydrolysis of H. tuberosus with 0.3 N of sulfuric acid resulted in fructose yield of 0.51 g g−1 and glucose yield of 0.14 g g−1 at 80 °C in 1 h. Proteolysis of algal residue with 2 N of sulfuric acid resulted in amino acid yield of 0.38 g g−1 at 110 °C in 24 h. To simplify the production of hydrolysates, the hydrolysis conditions were combined to establish a tailored sequential hydrolysis process. Then, simultaneous neutralization and salt removal of the hydrolysates were done with barium hydroxide. Consequently, cultivation of Ettlia sp. with the diluted sequential hydrolysate of H. tuberosus and LEA successfully achieved 5.90 g L−1 d−1 of biomass productivity and 1.42 g L−1 d−1 of lipid productivity with Ettlia sp. [ABSTRACT FROM AUTHOR]

[Display omitted] • Whole-cell biomass utilization was completed with newly integrated biorefinery. • Hydrolysis yielded 89 % of total carbohydrate within 4 min using optimized process. • Volume reduction decreased solvent usage by 75 % and recovered all oil within 20 min. • Bioethanol yield of sole hydrolysate was comparable with conventional YPD medium. • Newly developed process decreased OPEX and CAPEX by 82.7 % and 70.1 %, respectively. Microalgae are a promising source of biomass for the production of biofuels, but their use is currently infeasible because of changes in the biofuel market and the lack of an optimized refinery process. The biomass of oleaginous microalgae consists of up to 50% lipids and 50% carbohydrates and proteins, so the biorefinery process must be able to use whole cell biomass. In this work, an alternative process to the standard biorefinery approach of lipid extraction and treatment of lipid-extracted residue was developed and tested using oleaginous Chlorella sp. ABC-001. First, a hydrolysis process was optimized to achieve cell lysis. At 170 °C and 0.1 N H 2 SO 4 , 89.0% of total carbohydrates were hydrolyzed to monosaccharides within 4 min. Then, a volume reduction process (brief and mild centrifugation) was applied to reduce the extraction time and the use of organic solvent. By reducing the aqueous phase from the hydrolysate, all lipids were extracted by vortexing with just a quarter of the original volume (10 mL) within 20 min. Finally, after extraction the aqueous phase was utilized as the sole growth medium for ethanol production by Saccharomyces cerevisiae KL17. The resulting ethanol yield was 0.39 g ethanol/g sugar, comparable to that achieved using conventional yeast-peptone-dextrose (YPD) medium (0.41 g ethanol/g sugar). Furthermore, relative to the conventional process, techno-economic analysis confirmed that the newly developed process decreased operational expenditures by 82.7% and decreased capital expenditures by 70.1%. [ABSTRACT FROM AUTHOR]