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Second, yeast can overcome the growth deficiency caused by altered sterol metabolism through repeated culture. By employing metabolic engineering, strain evolution, fermentation engineering, and pathway reconstitution, we were able to reconstruct the multienzyme pathways for the synthesis of a set of phytosterols campesterol (∼7 mg/L), β-sitosterol (∼2 mg/L), 22-hydroxycampesterol (∼1 mg/L), and 22-hydroxycampest-4-en-3-one (∼4 mg/L). This work identified and addressed some of the technical bottlenecks in phytosterol-derived pathway reco