Display omitted •Main carotenoids from fungi and microalgae in past few years were reviewed.•Carotenoids production could be affected by combined stress and co-cultivation.•Green extraction methods can maintain biological activity of carotenoids.•Synthetic biology and bioinformatics were the key tools in the future research. The demand for carotenoids from natural sources obtained by biological extraction methods is increasing with the development of biotechnology and the continued awareness of food safety. Natural plant-derived carotenoids have a relatively high production cost and are affected by the season, while microbial-derived carotenoids are favored due to their natural, high-efficiency, low production cost, and ease of industrialization. This article reviewed the following aspects of natural carotenoids derived from microorganisms: (1) the structures and properties of main carotenoids; (2) fungal and microalgal sources of the main carotenoids; (3) influencing factors and modes of improvement for carotenoids production; (4) efficient extraction methods for carotenoids; and (5) the commercial value of carotenoids. This review provided a reference and guidance for the development of natural carotenoids derived from microorganisms.