This paper investigates the use of Li-LSX-zeolite catalyst over three regeneration cycles in presence of non-treated and acid pre-treated Isochrysis sp. microalgae. The spent and regenerated catalysts were characterised by surface analysis, elemental analysis (EA), SEM-EDS, and XRD to correlate their properties with the bio-oil yield and quality. The acid pre-treatment removed alkali metals, reducing gas yield in favour of bio-oil, but, at the same time, led to catalyst deactivation by fouling. Differently, the non-treated microalgae resulted in a bio-oil enriched in C and H and depleted in O, compared to the pre-treated ones, denoting higher deoxygenation activity. After 3 pyrolysis/regeneration cycles, the analyses suggest that there are no major changes on catalyst using non-treated microalgae. Regeneration at 700 °C has been shown to be able to remove most of the coke without damaging the Li-LSX zeolite structure. In summary, Li-LSX zeolite was effective in maintaining deoxygenation activity over three cycles in the pyrolysis of non-treated Isochrysis microalgae, while the algae pre-treatment with sulphuric acid was detrimental on the catalyst activity.