A Two-Step Process for Improved Biomass Production and Non-Destructive Astaxanthin and Carotenoids Accumulation in <i>Haematococcus pluvialis</i>

Carotenoids extracted from microalgae have a considerable economic interest in numerous high-value markets. Natural astaxanthin has gained much interest in its powerful antioxidant properties, however, its commercial-scale production is still challenging. In this study, a simple and economical way to cultivate Haematococcus pluvialis (CCAP 34/1D) by a two-step process was investigated by exploring alternative strategies to maximise algal growth and astaxanthin yield. During the first step—improving biomass production—four nitrogen sources were tested (NH4Cl, NaNO3, Fe(III)NO3, and urea). The second step—carotenogenesis induction—was achieved by using a mix of moderate stressors that worked in synergy (i.e., mild light, nitrogen limitation, the addition of sodium acetate at 0.25% w/v), thereby minimising potential losses of the accumulated biomass caused, for example, by photobleaching or nitrogen starvation. Results showed that urea was the nitrogen source, allowing the highest cell density and growth rate. In terms of carotenogenesis induction, the use of mild stressors resulted in three out of four treatments having a relative increase in cell number (13.8–26.7%) and a concomitant increase in astaxanthin yield. Simple low-cost strategies, such as small adjustments to media recipes and synergism between mild stressors, could bring a disproportionate effect on the future successes of making algal biotechnology a widespread reality