MELiSSA (Microecological Life Support System Alternative) is a developing technology for regenerative life support to enable long-term human missions in Space and has developed a demonstration Pilot Plant. One of the components of the MELiSSA Pilot Plant system is an 83L external loop air-lift photobioreactor (PBR) where Limnospira indica (previously named Arthrospira sp. PC8005) is axenically cultivated in a continuous operation mode for long-periods. Its mission is to provide O2 and consume CO2 while producing edible material. Biological and process characterization of this PBR is performed by analysing the effect of two main variables, dilution rate (D) and PFD (Photon Flux Density) illumination. A maximum oxygen productivity (urn:x-wiley:17517915:media:mbt213882:mbt213882-math-0001) of 1.35 mmol l−1 h−1 is obtained at a D of 0.025 h−1 and PFD of 930 µmol m−2 s−1. Photoinhibition can occur when a 1 g l−1 cell density culture is exposed to PFD higher than 1700 µmol m−2 s−1. This process is reversible if the illumination is returned to dim light (150 µmol m−2 s−1), proving the cell adaptability and capacity to respond at different illumination conditions. Influence of light intensity in cell composition is also described. Specific photon flux density (qPFD) has a direct effect on phycobiliproteins and chlorophyll content causing a decrease of 62.5% and 47.8%, respectively, when qPFD increases from 6.1 to 19.2 µmol g−1 s−1. The same trend is observed for proteins and the opposite for carbohydrate content. Morphological and spiral structural features of L. indica are studied by confocal microscopy, and size distribution parameters are quantified. A direct effect between trichome width and CDW/OD ratio is observed. Changes in size distribution are not correlated with environmental factors, further confirms the adaptation capacity of the cells. The systematic analysis performed provides valuable insights to understand the key performance criteria of continuous culture in air-lift PBRs.