Carotenoids are valuable lipophilic compounds with commercial applications in food, animal feed, cosmetics, and nutraceutrical industry. They are found widespread in nature, as they can be naturally synthesized by all photosynthetic organisms, along with some non-photosynthetic organisms as well. However, natural carotenoid extraction is labor-intensive and costly. By comparison, synthetic carotenoids derived from petrochemical precursors are often deemed unsuitable by regulatory agencies for application in drug, food, and cosmetics. Turning to microbial production have long been suggested as an alternative to the both sources by offering better sustainability, safety and economics. In this thesis, multiple strategies were explored to produce β-carotene or zeaxanthin in oleaginous yeast Yarrowia lipolytica. Introduction of carotenoid synthesis enzymes resulted in residual precursors, leading to the exploration of multiple corrective strategies such as pathway flux optimization, redox balancing and cellular compartmentalization. Chapter 3 outline the engineering strategies that enabled the non-producing Y. lipolytica strain to produce β-carotene. After introduction of heterolgous β-carotene pathway, various genes in upstreme pathway were overexpressed. Lycopene accumulation following the increased carotenoid production level was eliminated using previous reported mutation in the CarRP gene. Two approaches to redirect the central carbon pathway were tested to promote NADPH regeneration and reduce ATP expenditure. Lastly flux to lipid production pathway was increased to facilitate more β-carotene accommodation in lipid droplet (LD), while ensuring enough acetyl-CoA pool for carotenoid production. In chapter 4, a β-carotene hydroxylase gene was introduced into a β-carotene producing strain from the previous chapter, in order to produce zeaxanthin. The best host-compatible enzyme, PaCrtZ was selected after comparison with various homologs. After finding an enzyme mutant of PaCrtZ from co-evolutionary analysis, various approaches were taken to improve zeaxanthin production. From leucine phototrophy, oxygen and pH control, redox cofactor engineering, several conditions that improved zeaxanthin synthesis were found. Lastly, chapter 5 describes strategies that may promote accomodation of PaCrtZ mutant enzyme and zeaxanthin in cell. Targeted relocation of the PaCrtZ mutant enzyme to ER, peroxisome and LD was performed and analyzed. ER and peroxisome membrane expansion were explored. Strategy that may shift LD morphology toward small and numerous were demonstrated.