Dunaliella salina is a halotolerant, photoautotrophic marine microalga and one of the richest sources of natural carotenoids but also shows potential as a novel food source with high protein quality. This study sought to optimise the production of biomass, protein and amino acids from D. salina, alongside carotenoids using a two-stage cultivation approach based on the use of light of different intensities and quality, i.e., white, red and blue LED light. In stage 1, four white LED light intensities were tested. In stage 2, the same four light intensities from either blue or red LEDs were applied once exponential growth ceased and cells reached the stationary phase under white LED light in stage 1. Remarkably, both biomass concentration and biomass productivity showed a 1.3–1.7-fold increase in stage 2, without medium replenishment, while protein concentration and protein productivity showed an ~1.1-fold increase. The amino acid content and amino acid index remained unchanged from stage 1 to stage 2, and minimum difference was found across different light intensities. Overall, D. salina delivered so-called high protein quality, with an essential amino acid index (EAAI) of 0.99, and red light, which has previously been shown to increase carotenoid production, boosted further biomass production over and above white light, at all light intensities tested.