Insights into the physiology of Chlorella vulgaris cultivated in sweet sorghum bagasse hydrolysate for sustainable algal biomass and lipid production

Abstract

Supplementing cultivation media with exogenous carbon sources enhances biomass and lipid production in microalgae. Utilization of renewable organic carbon from agricultural residues can potentially reduce the cost of algae cultivation, while enhancing sustainability. In the present investigation a medium was developed from sweet sorghum bagasse for cultivation of Chlorella under mixotrophic conditions. Using response surface methodology, the optimal values of critical process parameters were determined, namely inoculum cell density (O.D.₇₅₀) of 0.786, SSB hydrolysate content of the medium 25% v/v, and zero medium salinity, to achieve maximum lipid productivity of 120 mg/L/d. Enhanced biomass (3.44 g/L) and lipid content (40% of dry cell weight) were observed when the alga was cultivated in SSB hydrolysate under mixotrophic conditions compared to heterotrophic and photoautotrophic conditions. A time course investigation revealed distinct physiological responses in terms of cellular growth and biochemical composition of C. vulgaris cultivated in the various trophic modes. The determined carbohydrate and lipid profiles indicate that sugar addition to the cultivation medium boosts neutral lipid synthesis compared to structural lipids, suggesting that carbon flux is channeled towards triacylglycerol synthesis in the cells. Furthermore, the fatty acid profile of lipids extracted from mixotrophically grown cultures contained more saturated and monosaturated fatty acids, which are suitable for biofuel manufacturing. Scale-up studies in a photobioreactor using SSB hydrolysate achieved a biomass concentration of 2.83 g/L consisting of 34% lipids and 26% carbohydrates. These results confirmed that SSB hydrolysate is a promising feedstock for mixotrophic cultivation of Chlorella and synthesis of algal bioproducts and biofuels.

Figure 1

Dry cell weight (DCW), total lipid content, and total lipid productivity of Chlorella 395, Chlorella 1230, and Chlorella 26 on the 10th day of batch cultivation in BBM medium (BBM) vs. SSB hydrolysate (SBB). (*p < 0.05; **p < 0.01; ns: not significant).

Figure 2

Response surface plots (3D) showing the effect of (a) salinity and hydrolysate; (b) OD₇₅₀ and salinity; and (c) hydrolysate and OD₇₅₀ on lipid productivity of Chlorella 395.

Figure 3

Growth performance in terms of OD₇₅₀ and dry cell weight of Chlorella 395 cultivated in various growth media and trophic modes. Lines represent OD_750nm, whereas bars represent DCW.

Figure 4

Temporal changes in pH, nitrate, phosphate, glucose, and xylose concentration of the culture media of Chlorella 395 cultivated (a) autotrophically; (b) mixotrophically in pure sugars; (c) mixotrophically in SSB hydrolysate; and (d) heterotrophically in SSB hydrolysate.

Figure 5

Changes in (a) total lipid content and productivity; (b) total carbohydrate content and productivity; (c) total protein content and productivity; and (d) photosynthetic pigment content of Chlorella 395 cultivated in various growth media and trophic modes. Lines represent productivity, whereas bars represent cellular content (%).

Figure 6

(a) Relative changes in FAME composition; (b) changes in polar lipid concentration; and (c) alterations in cellular carbohydrate content of Chlorella 395 cultivated in various growth media and trophic modes.