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Open cultivation systems are the most economic option for microalgal biomass production but their harvesting cost, bacterial invasion, and water usage are still deterring large-scale applications. Therefore, in this study, low-cost techniques for harvesting, bacterial disinfection, and medium recycling were sequentially implemented for Ettlia sp. For harvesting, the employed auto-flocculation harvested 96% of algae but was not able to remove bacteria that invaded during cultivation. For this reason, hydrodynamic treatment was applied as a bacterial disinfection method for medium recycling, and treated up to 100% of the bacteria in the supernatant. Afterward, the supernatant was replenished with nutrients and reused to grow microalgae. In medium-recycled cultivation, auto-flocculation recycled (AR) medium showed the highest final biomass concentration of 3.84 g L−1, while the centrifugation recycled (CR) medium and the fresh medium (FM) resulted in 3.78 and 3.67 g L−1, respectively. In addition, the lipid content of the AR group was highly increased to 20.4%, resulting in the highest lipid productivity of 0.11 g L−1 d−1. Moreover, an analysis of fatty acid methyl ester compositions confirmed that 76.6% of lipid from the AR group were mainly composed of C16-C18, which are suitable for biodiesel. Consequently, the results showed that the implementation of low-cost techniques not only reduced process costs and water usage but also enhances both the biomass and lipid productivities of the species. [Display omitted] • Low-cost harvesting, bacteria control, and medium recycling were combined. • Auto-flocculation harvested microalgae but left bacteria in the supernatant. • Hydrodynamic cavitation disinfected bacteria with low-energy consumption. • Biomass productivity of auto-flocculation recycled medium was slightly increased. • Auto-flocculation recycled medium resulted in 1.6-fold higher lipid productivity. [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]

Lipid production in microalgae under nitrogen (N) starved condition can be enhanced by excess phosphorus (P) supply in the second stage of two-stage cultivation strategy. However, implementing two-stage cultivation is difficult in large-scale cultivation system as it requires high energy of transferring large algal biomass from first stage to second stage. To address this problem, we have optimized a continuous two-stage (CTS) cultivation strategy using Chlorella sp. HS2, where nitrogen in the growth environment is depleted naturally via consumption. To enhance both biomass and lipid productivity this strategy explored supplementation of additional P from 50% to 2500% of the initial concentration at the start of N-limited second stage of growth. The results of the optimization study in photobioreactor (PBR) showed that supplementing 500% of initial P and 100% of initial other nutrients (O) (N 0 –P 500 –O 100) on 5th day showed the maximum biomass productivity of 774.4 mg L−1 d−1. It was observed that Chlorella sp. HS2 grown in PBR yielded higher biomass (3.8 times), lipid (6.1 times) and carbohydrate (5.5 times) productivity in comparison to the open raceway ponds (ORP) study, under optimum nutrient and carbon supply condition. The maximum lipid (289.6 mg L−1 d−1) and carbohydrate (219.2 mg L−1 d−1) productivities were obtained in TPBR-3, which were 1.9 and 1.3 times higher than that of TPBR-2 (+ve control) and 9.6 and 3.7 times higher than that of TPBR-1 (-ve control), respectively. Fatty acid mainly composed of C 16 /C 18 (84.5%–85.7%), which makes the microalgal oil suitable for biofuel production. This study concluded that feeding excess amount of P is an effective and scalable strategy to improve the biomass and lipid productivity of CTS cultivation. Image 1 • Chlorella sp. HS2 was grown in a CTS cultivation process with optimum carbon supply. • Biomass growth was maximum with 500% excess P supply (N 0 –P 500 –O 100) in 2nd stage. • Both TPBR-3 and TORP-3 achieved the enhanced lipid and carbohydrate productivity. • Provide a strategy for high P utilization and maximizing the biofuel production. [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]