Engineering Yarrowia lipolytica for poly-3-hydroxybutyrate production
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Strains of Yarrowia lipolytica were engineered to express the poly-3-hydroxybutyrate (PHB) biosynthetic pathway. The genes for β-ketothiolase, NADPH-dependent acetoacetyl-CoA reductase, and PHB synthase were cloned and inserted into the chromosome of Y. lipolytica. In shake flasks, the engineered strain accumulated PHB to 1.50 and 3.84% of cell dry weight in complex medium supplemented with glucose and acetate as carbon source, respectively. In fed-batch fermentation using acetate as sole carbon source, 7.35 g/l PHB (10.2% of cell dry weight) was produced. Selection of Y. lipolytica as host for PHB synthesis was motivated by the fact that this organism is a good lipids producer, which suggests robust acetyl-CoA supply also the precursor of the PHB pathway. Acetic acid could be supplied by gas fermentation, anaerobic digestion, and other low-cost supply route.
KeywordsPoly-3-hydroxybutyrate PHB Metabolic engineering phaCAB Acetate Yarrowia lipolytica
We thank Ms. Xue-Mei Che of the School of Life Sciences of Tsinghua University for the assistance of PHB molecular weight assays. This research was financially supported by Grants from the Department of Energy (DE-SC0008744). ZJL was funded by the National Natural Science Foundation of China (21476014 and 31100025).
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