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Applied Microbiology and Biotechnology

, Volume 100, Issue 8, pp 3781–3798 | Cite as

Functional overexpression and characterization of lipogenesis-related genes in the oleaginous yeast Yarrowia lipolytica

  • Andrew M. Silverman
  • Kangjian Qiao
  • Peng Xu
  • Gregory StephanopoulosEmail author
Bioenergy and biofuels

Abstract

Single cell oil (SCO) is an attractive energy source due to scalability, utilization of low-cost renewable feedstocks, and type of product(s) made. Engineering strains capable of producing high lipid titers and yields is crucial to the economic viability of these processes. However, lipid synthesis in cells is a complex phenomenon subject to multiple layers of regulation, making gene target identification a challenging task. In this study, we aimed to identify genes in the oleaginous yeast Yarrowia lipolytica whose overexpression enhances lipid production by this organism. To this end, we examined the effect of the overexpression of a set of 44 native genes on lipid production in Y. lipolytica, including those involved in glycerolipid synthesis, fatty acid synthesis, central carbon metabolism, NADPH generation, regulation, and metabolite transport and characterized each resulting strain’s ability to produce lipids growing on both glucose and acetate as a sole carbon source. Our results suggest that a diverse subset of genes was effective at individually influencing lipid production in Y. lipolytica, sometimes in a substrate-dependent manner. The most productive strain on glucose overexpressed the diacylglycerol acyltransferase DGA2 gene, increasing lipid titer, cellular content, and yield by 236, 165, and 246 %, respectively, over our control strain. On acetate, our most productive strain overexpressed the acylglycerol-phosphate acyltransferase SLC1 gene, with a lipid titer, cellular content, and yield increase of 99, 91, and 151 %, respectively, over the control strain. Aside from genes encoding enzymes that directly catalyze the reactions of lipid synthesis, other ways by which lipogenesis was increased in these cells include overexpressing the glycerol-3-phosphate dehydrogenase (GPD1) gene to increase production of glycerol head groups and overexpressing the 6-phosphogluconolactonase (SOL3) gene from the oxidative pentose phosphate pathway to increase NADPH availability for fatty acid synthesis. Taken together, our study demonstrates that the overall kinetics of microbial lipid synthesis is sensitive to a wide variety of factors. Fully optimizing a strain for single cell oil processes could involve manipulating and balancing many of these factors, and, due to mechanistic differences by which each gene product investigated here impacts lipid synthesis, there is a high likelihood that many of these genes will work synergistically to further increase lipid production when simultaneously overexpressed.

Keywords

Lipogenesis Metabolic engineering Gene expression Fermentation 

Notes

Acknowledgments

This material is based upon work supported by the US Department of Energy, Office of Science, Office of Biological and Environmental Research, Genomic Science program, under Award Number DE-SC0008744.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Human and animals studies

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2016_7376_MOESM1_ESM.pdf (408 kb)
ESM 1 (PDF 407 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Andrew M. Silverman
    • 1
  • Kangjian Qiao
    • 1
  • Peng Xu
    • 1
  • Gregory Stephanopoulos
    • 1
    Email author
  1. 1.Department of Chemical EngineeringMassachusetts Institute of TechnologyCambridgeUSA

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