Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Understanding and eliminating the detrimental effect of endogenous thiamine auxotrophy on metabolism of the oleaginous yeast Yarrowia lipolytica

Caleb Walker, Seunghyun Ryu, Richard J. Giannone, Sergio Garcia, Cong T. Trinh
doi: https://doi.org/10.1101/753525
Caleb Walker
Department of Chemical and Biomolecular Engineering, The University of Tennessee Knoxville, TN 37996
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Seunghyun Ryu
Department of Chemical and Biomolecular Engineering, The University of Tennessee Knoxville, TN 37996
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Richard J. Giannone
Chemical Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Sergio Garcia
Department of Chemical and Biomolecular Engineering, The University of Tennessee Knoxville, TN 37996
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Cong T. Trinh
Department of Chemical and Biomolecular Engineering, The University of Tennessee Knoxville, TN 37996
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: ctrinh@utk.edu
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Preview PDF
Loading

ABSTRACT

Thiamine is an essential vitamin that functions as a cofactor for key enzymes in carbon and energy metabolism for all living cells. While most plants, fungi and bacteria can synthesize thiamine de novo, the oleaginous yeast, Yarrowia lipolytica, cannot. In this study, we used proteomics together with physiological characterization to understand key metabolic processes influenced and regulated by thiamine availability and identified the genetic basis of thiamine auxotrophy in Y. lipolytica. Specifically, we found thiamine depletion results in decreased protein abundance of the lipid biosynthesis pathways and energy metabolism (i.e., ATP synthase), attributing to the negligible growth and poor sugar assimilation observed in our study. Using comparative genomics, we identified the missing gene scTHI13, encoding the 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate synthase for the de novo thiamine synthesis in Y. lipolytica, and discovered an exceptional promoter, P3, that exhibits strong activation or tight repression by low and high thiamine concentrations, respectively. Capitalizing on the strength of our thiamine-regulated promoter (P3) to express the missing gene, we engineered the first thiamine-prototrophic Y. lipolytica reported to date. By comparing this engineered strain to the wildtype, we unveiled the tight relationship linking thiamine availability to lipid biosynthesis and demonstrated enhanced lipid production with thiamine supplementation in the engineered thiamine-prototrophic Y. lipolytica.

IMPORTANCE Thiamine plays a crucial role as an essential cofactor for enzymes in carbon and energy metabolism of all living cells. Thiamine deficiency has detrimental consequences on cellular health. Yarrowia lipolytica, a non-conventional oleaginous yeast with broad biotechnological applications, is a native thiamine auxotroph, whose effects on cellular metabolism are not well understood. Therefore, Y. lipolytica is an ideal eukaryotic host to study thiamine metabolism, especially as mammalian cells are also thiamine-auxotrophic and thiamine deficiency is implicated in several human diseases. This study elucidates the fundamentals of thiamine deficiency on cellular metabolism of Y. lipolytica and identifies genes and novel thiamine-regulated elements that eliminate thiamine auxotrophy in Y. lipolytica. Furthermore, discovery of thiamine-regulated elements enables development of thiamine biosensors with useful applications in synthetic biology and metabolic engineering.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
Back to top
PreviousNext
Posted September 04, 2019.
Download PDF

Supplementary Material

Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Understanding and eliminating the detrimental effect of endogenous thiamine auxotrophy on metabolism of the oleaginous yeast Yarrowia lipolytica
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
Share
Understanding and eliminating the detrimental effect of endogenous thiamine auxotrophy on metabolism of the oleaginous yeast Yarrowia lipolytica
Caleb Walker, Seunghyun Ryu, Richard J. Giannone, Sergio Garcia, Cong T. Trinh
bioRxiv 753525; doi: https://doi.org/10.1101/753525
Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Understanding and eliminating the detrimental effect of endogenous thiamine auxotrophy on metabolism of the oleaginous yeast Yarrowia lipolytica
Caleb Walker, Seunghyun Ryu, Richard J. Giannone, Sergio Garcia, Cong T. Trinh
bioRxiv 753525; doi: https://doi.org/10.1101/753525

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Microbiology
Subject Areas
All Articles
  • Animal Behavior and Cognition (1545)
  • Biochemistry (2500)
  • Bioengineering (1757)
  • Bioinformatics (9729)
  • Biophysics (3929)
  • Cancer Biology (2990)
  • Cell Biology (4235)
  • Clinical Trials (135)
  • Developmental Biology (2653)
  • Ecology (4129)
  • Epidemiology (2033)
  • Evolutionary Biology (6933)
  • Genetics (5243)
  • Genomics (6532)
  • Immunology (2208)
  • Microbiology (7012)
  • Molecular Biology (2784)
  • Neuroscience (17412)
  • Paleontology (127)
  • Pathology (432)
  • Pharmacology and Toxicology (712)
  • Physiology (1068)
  • Plant Biology (2516)
  • Scientific Communication and Education (647)
  • Synthetic Biology (835)
  • Systems Biology (2699)
  • Zoology (439)