INTRODUCTION: The marine microalga is prolific producer of fucoxanthin, which is a xanthophyll carotenoid with substantial global market value boasting extensive applications in the food, nutraceutical, pharmaceutical, and cosmetic industries. This study presented a novel integrated experimental approach coupled with machine learning (ML) models to predict the fucoxanthin content in by altering the type and concentration of phytohormone supplementation, thus overcoming the multiple methodological limitations of conventional fucoxanthin quantification. METHODS: A novel integrated experimental approach was developed, analyzing the effect of varying phytohormone types and concentrations on fucoxanthin production in . Morphological analysis was conducted to assess changes in microalgal structure, while growth rate and fucoxanthin yield correlations were explored using statistical analysis and machine learning models. Several ML models were employed to predict fucoxanthin content, with and without hormone descriptors as variables. RESULTS: The findings revealed that the Random Forest (RF) model was highly significant with a high of 0.809 and of 0.776 when hormone descriptors were excluded, and the inclusion of hormone descriptors further improved prediction accuracy to of 0.839, making it a useful tool for predicting the fucoxanthin yield. The model that fitted the experimental data indicated methyl jasmonate (0.2 mg/L) as an effective phytohormone. The combined experimental and ML approach demonstrated rapid, reliable, and cost-efficient prediction of fucoxanthin yield. DISCUSSION: This study highlights the potential of machine learning models, particularly Random Forest, to optimize parameters influencing microalgal growth and fucoxanthin production. This approach offers a more efficient alternative to conventional methods, providing valuable insights into improving fucoxanthin production in microalgal cultivation. The findings suggest that leveraging diverse ML models can enhance the predictability and efficiency of fucoxanthin production, making it a promising tool for industrial applications.