Acetate is an important renewable carbon source that has been widely applied in the biosynthesis of high-value chemicals.(R)-3-hydroxybutyrate[(R)-3-HB], as a crucial chiral intermediate in the food and pharmaceutical industries, holds broad application potential.However, microbial synthesis of (R)-3-HB from acetate as the sole substrate has remained limited by low titers.In this study, we aimed to construct an engineered Yarrowia lipolytica strain capable of efficiently converting acetate into (R)-3-HB.First, a high acetate-tolerant strain, YL305, was obtained through strain library screening, and a CRISPR-based gene editing toolkit was established.Subsequently, optimal (R)-3-HB biosynthetic modules were developed by screening for efficient combinations of acetoacetyl-CoA reductases (PhaBs) and thioesterases.By further optimizing key gene copy numbers, enhancing acetyl-CoA supply, and disrupting competing (R)-3-HB-CoA bypass pathways, the engineered strain achieved a titer of 377.86 mg/L (R)-3-HB in shake flask cultivation.Moreover, during fed-batch fermentation in a 5 L bioreactor with acetate as the sole carbon source, (R)-3-HB production reached at 5.50 g/L, representing the highest level reported to date in Y.lipolytica.This work demonstrates the application potential of Y.lipolytica in acetate valorization through systematic metabolic engineering, providing an effective microbial chassis and strategy for the efficient synthesis of acetate-derived chemicals.
LI Zichen
,
LIU Qi
,
DAI Zongjie
,
WANG Qinhong
. Metabolic engineering of acetate-tolerant Yarrowia lipolytica for efficient production of (R)-3-hydroxybutyric acid[J]. Food and Fermentation Industries, 2026
, 52(10)
: 166
-174
.
DOI: 10.13995/j.cnki.11-1802/ts.044375
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