Microbially produced lipids have attracted attention for their environmental benefits and commercial value. We have combined lipid pathway engineering in Saccharomyces cerevisiae yeast with bioprocess design to improve productivity and explore barriers to enhanced lipid production. Initially， individual gene expression was tested for impact on yeast growth and lipid production. Then， two base strains were prepared for enhanced lipid accumulation and stabilization steps by combining DGAT1， ΔTgl3 with or without Atclo1， which increased lipid content ~ 1.8-fold but reduced cell viability. Next， fatty acid (FA) biosynthesis genes Ald6-SEACS alone or with ACC1** were co-expressed in base strains， which significantly improved lipid content (8.0% DCW， 2.6-fold than control)， but severely reduced yeast growth and cell viability. Finally， a designed two-stage process convincingly ameliorated the negative effects， resulting in normal cell growth， very high lipid productivity (307 mg/L， 4.6-fold above control) and improved cell viability.