Single-use food packaging contributes towards almost one-third of the global plastic waste. In an attempt to minimize environmental impacts, sustainable alternatives need to be developed. Seaweed is a renewable resource that can be utilized for packaging development for plastic replacement. Gracilaria is one of the most widely-cultivated algae, used in food and pharmaceutical industries as a valuable source of agar. Studies have succeeded in developing biopolymer plastics from Gracilaria containing high levels of antioxidants. Antioxidant properties are essential in biopackaging as they are beneficial to packaged food by limiting lipid oxidation, which is one the main causes of the food spoilage. This study focuses on the cultivation of Gracilaria vermiculophylla and optimizing its antioxidant properties for further use as an edible film for food packaging. Antioxidant compounds are synthesized in seaweeds as a response to oxidative stress that often occurs due to harsh environmental fluctuations. Therefore, antioxidant activity of the seaweed can be modified through exposure to different stress conditions. Three consecutive experiments were conducted in this study to better understanding on how different levels of light irradiance, differences in light : dark cycles, exposure to UVA radiation, hypersalinity, desiccation and the duration of exposure can affect the antioxidant profile of Gracilaria vermiculophylla. High light intensity increased the antioxidant levels of G. vermiculophylla, reaching a maximum of 33.45%. Moreover, the combination of the high light irradiances and UVA enhanced the antioxidant activity and had a positive survival effect on G. vermiculophylla. The study showed that increased antioxidant activity could be achieved after 3 days of exposure to different cultivating conditions. These findings may be beneficial to industrial scale seaweed cultivation, where favorable antioxidant levels could be reached shortly prior to harvesting.