The escalating global burden of diabetes underscores the urgent need for sustainable insulin production. This study explores the potential of Arthrospira platensis SPKY1 as an alternative source of insulin, particularly pertinent in regions with high diabetes prevalence like India. Through comprehensive experimentation, factors influencing insulin production in A. platensis SPKY1 are investigated, including growth media composition, pH levels, light conditions, greenhouse cultivation, water types and carbon sources. Results reveal those higher concentrations of specific growth media components, such as NaHCO₃, NaNO₃, NaCl, K₂SO₄, and K₂HPO₄, correlate with increased insulin production. Among these components, K₂SO₄ at a concentration of 1.4 g L^-1 showed the highest insulin production, reaching 27.5 µg g^-1. Additionally, the study evaluated the impact of various pH levels, finding that pH 10.0 yielded optimal insulin production, with a peak of 21.3 µg g^-1. Blue light exposure stimulated the most significant increase in insulin production, with levels ranging from 5.4 to 25.1 µg g^-1. Additionally, enriched seawater proved more effective than regular medium for insulin production. The study also demonstrated that glucose proved to be the optimal carbon source, with insulin production reaching 29.4 µg g^-1. The study determines the optimal growth conditions of A. platensis SPKY1 for insulin production on a pilot scale.