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Evolutionary engineering improved D-Glucose/Xylose cofermentation of yarrowia lipolytica

Authors
  • Zhou, L
  • Wen, Z
  • Wang, Z
  • Zhang, Y
  • Ledesma-Amaro, R
  • Jin, M
Publication Date
Jun 01, 2020
Source
UPCommons. Portal del coneixement obert de la UPC
Keywords
License
Unknown

Abstract

The recombinant strain Y. lipolytica XYL+ can utilize xylose to produce microbial lipids efficiently. However, its xylose uptake is severely delayed in the presence of d-glucose. In this study, adaptive laboratory evolution (ALE) of strains in the medium containing xylose and d-glucose analogue 2-deoxyglucose (dG) is performed. After four stages of evolution over a total of 64 days, for the first time, we obtained an Y. lipolytica strain (yl-XYL+*04*10) that can cometabolize xylose and glucose, with diminished glucose repression. Xylose uptake kinetics showed that it could efficiently utilize xylose in the presence of 10 g/L of dG or d-glucose. Transcriptional profiling analysis revealed that the relative expression level of xylose-specific transporter genes (encoded by YALI0_C04730g and YALI0_D00363g) was significantly upregulated. In addition, missense mutations N373T and G270A in YALI0_E23287g (encoding a d-glucose transporter) and YALI0_E15488g (encoding a hexokinase) respectively, were found. It implied that these are important gene targets responsible for improved xylose utilization in evolved Y. lipolytica. Our work provides a new approach for breeding Y. lipolytica and paved the way for future pentose metabolic engineering.

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