The heterotrophic cultivation of microalgae has a number of notable advantages, which include allowing high culture density levels as well as enabling the production of biomass in consistent and predictable quantities. In this study, the full potential of Chlorella sp. HS2 is explored through optimization of the parameters for its heterotrophic cultivation. First, carbon and nitrogen sources were screened in PhotobioBox. Initial screening using the Plackett-Burman design (PBD) was then adopted and the concentrations of the major nutrients (glucose, sodium nitrate, and dipotassium phosphate) were optimized via response surface methodology (RSM) with a central composite design (CCD). Upon validation of the model via flask-scale cultivation, the optimized BG11 medium was found to result in a three-fold improvement in biomass amounts, from 5.85 to 18.13 g/L, in comparison to a non-optimized BG11 medium containing 72 g/L glucose. Scaling up the cultivation to a 5-L fermenter resulted in a greatly improved biomass concentration of 35.3 g/L owing to more efficient oxygenation of the culture. In addition, phosphorus feeding fermentation was employed in an effort to address early depletion of phosphate, and a maximum biomass concentration of 42.95 g/L was achieved, with biomass productivity of 5.37 g/L/D.

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.

Microalgal biomass was hydrolyzed using a solid acid catalyst with the aid of liquid acid. The use of solid acid as the main catalyst instead of liquid acid was to omit subsequent neutralization and/or desalination steps, which are commonly required in using the resulting hydrolysates for microbial fermentation. The hydrolysis of 10 g/L of lipid‐extracted Chlorella vulgaris containing 12.2% carbohydrates using 7.6 g/L Amberlyst 36 and 0.0075 N nitric acid at 150°C resulted in 1.08 g/L of mono‐sugars with a yield of 88.5%. For hydrolysis of higher concentrations of the biomass over 10 g/L, the amount of Amberlyst 36 needed to be increased in proportion to the biomass concentration to maintain similar levels of hydrolysis performance. Increasing the solid acid concentration protected the surface of the solid acid from being severely covered by cell debris during the reaction. A hydrolysate of lipid‐extracted C. vulgaris 50 g/L was used, with no post‐treatment of desalination, for the cultivation of Klebsiella oxytoca producing 2,3‐butanediol. Cell growth in the hydrolysate was found to be almost the same as in the conventional medium with the same monosaccharide composition, confirming its fermentation compatibility. It was noticeable that the yield of 2,3‐butanediol with the hydrolysate was observed to be 2.6 times higher than that with the conventional medium. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2729, 2019

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