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C30 carotenoid biosynthetic genes of Staphylococcus aureus subsp. aureus were cloned and assembled in Escherichia coli. When dehydrosqualene synthase (CrtM) and dehydrosqualene desaturase (CrtN) were coexpressed in E .coli, 4,4’-diaponeurosporene was produced as a main carotenoid followed by fully conjugated 4,4’-diapolycopne. Directed evolution of CrtN generated 10 mutant CrtN clones showing altered activity. Some of the mutant clones produced 4,4’-diaponeurosporene exclusively and some produced 4,4’-diapolycopne dominantly when complemented with CrtM in E. coli. When the engineered 4,4’-diaponeurosporene and 4,4’-diapolycopne pathways were further extended with diaponeurosporene oxidase (CrtP), oxo-carotenoid 4,4’-diaponeurosporene-al was produced with a small amount of 4,4’-diaponeurosporene. Furthermore, the engineered C30 carotenoids pathways were successfully extended by C30 and C40 carotenoid pathway downstream enzymes resulting in several structures of C30 carotenoids.