Abstract
The present study aims to increase pyruvate production by engineering Yarrowia lipolytica through modifying the glycerol metabolic pathway. Results: Wild-type Yarrowia lipolytica (Po1d) was engineered to produce six different strains, namely ZS099 (by over-expressing PYK1), ZS100 (by deleting DGA2), ZS101 (by over-expressing DAK1, DAK2, and GCY1), ZS102 (by over-expressing GUT1 and GUT2), ZS103 (by over-expressing GUT1) and ZSGP (by over-expressing POS5 and deleting GPD2). Production of pyruvate from engineered and control strains was determined using high-performance liquid chromatography (HPLC). Subsequently, the fermentation conditions for producing pyruvate were optimized, including the amount of initial inoculation, the addition of calcium carbonate (CaCO3), thiamine and glycerol. Finally, for scaled-up purposes, a 20-L fermentor was used. It was observed that pyruvate production increased by 136% (8.55 g/L) in ZSGP strain compared to control (3.62 g/L). Furthermore, pyruvate production by ZSGP reached up to 110.4 g/L in 96 h in the scaled-up process. We conclude that ZSGP strain of Y. lipolytica can be effectively used for pyruvate production at the industrial level.
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Abbreviations
- DAK:
-
Dihydroxyacetone kinases
- GUT1:
-
Glycerol kinase
- GUT2:
-
3-Phosphoglycerol dehydrogenase
- GPD:
-
Glyceraldehyde-3-phosphate dehydrogenase
- GPP:
-
Glycerol-1-phosphatase
- GCY1:
-
Glycerol dehydrogenase
- Gly-3-p:
-
Glycerol-3- phosphate
- DHA:
-
1, 3-Dihydroxyacetone
- PYK1:
-
Pyruvate kinase
- POS5:
-
NADH kinase POS5
- TAG:
-
Triacylglycerol
- DGA:
-
Diacylglycerol acyltransferase
- DHAP:
-
Dihydroxyacetone phosphate
- MPC:
-
Mitochondrial pyruvate carrier
- GAP:
-
Glyceraldehyde-3-phosphate
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Acknowledgements
This study was financially supported by the Zhejiang Provincial Natural Science Foundation of China (Grant Nos. LQ18C010006 and LY19C010005), and National Natural Science Foundation of China (Grant Number: 31900497).
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Wang, S., Yang, Y., Yu, K. et al. Engineering of Yarrowia lipolytica for producing pyruvate from glycerol. 3 Biotech 12, 98 (2022). https://doi.org/10.1007/s13205-022-03158-7
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DOI: https://doi.org/10.1007/s13205-022-03158-7