Abstract: The organic-rich mudstones and dolostones of the Permian Fengcheng Formation (Fm.) are typically alkaline lacustrine source rocks, which are typified by impressively abundant β-carotane. Abundant β-carotane has been well acknowledged as an effective indicator of biological sources or depositional environments. However, the specific biological sources of β-carotane and the coupling control of biological sources and environmental factors on the enrichment of β-carotane in the Fengcheng Fm. remains obscure. Based on a comprehensive investigation of the bulk, molecular geochemistry, and organic petrology of sedimentary rocks and the biochemistry of phytoplankton in modern alkaline lakes, we proposed a new understanding of the biological precursors of β-carotane and elucidated the enrichment mechanism of β-carotane in the Fengcheng Fm. The results show that the biological precursors crucially control the enrichment of β-carotane in the Fengcheng Fm. The haloalkaliphilic cyanobacteria are the primary biological sources of β-carotane, which is suggested by a good positive correlation between the 2-methylhopane index, 7- + 8- methyl heptadecanes/Cmax, C29%, and β-carotane/Cmax in sedimentary rocks and the predominance of cyanobacteria with abundant β-carotene in modern alkaline lakes. The enrichment of β-carotane requires the reducing condition, and the paleoredox state that affects the enrichment of β-carotane appears to have a threshold. The paleoclimate conditions do not considerably impact the enrichment of β-carotane, but they have some influence on the water’s paleosalinity by affecting evaporation and precipitation. While it does not directly affect the enrichment of β-carotane in the Fengcheng Fm., paleosalinity does have an impact on the cyanobacterial precursor supply and the preservation conditions.