Urea and urine are a viable and cost-effective nitrogen source for Yarrowia lipolytica biomass and lipid accumulation
Yarrowia lipolytica is an industrial yeast that has been used in the sustainable production of fatty acid-derived and lipid compounds due to its high growth capacity, genetic tractability, and oleaginous properties. This investigation examines the possibility of utilizing urea or urine as an alternative to ammonium sulfate as a nitrogen source to culture Y. lipolytica. The use of a stoichiometrically equivalent concentration of urea in lieu of ammonium sulfate significantly increased cell growth when glucose was used as the carbon source. Furthermore, Y. lipolytica growth was equally improved when grown with synthetic urine and real human urine. Equivalent or better lipid production was achieved when cells are grown on urea or urine. The successful use of urea and urine as nitrogen sources for Y. lipolytica growth highlights the potential of using cheaper media components as well as exploiting and recycling non-treated human waste streams for biotechnology processes.
KeywordsYarrowia lipolytica Urea Urine Nitrogen Metabolism
The authors would like to acknowledge Dr. David Bruce for access to GC-FID and Dr. Rodrigo Martinez Duarte for access to BSL-2 biosafety cabinet for human urine experiments. The authors would also like to acknowledge Dr. Difeng Gao for creating the ∆pex10 strain that was used in this study.
This work was supported by an Early Career Faculty Award from NASA’s Space Technology Research Grants Program (#NNX15AU46G) to MAB, the US National Science Foundation (CBET-1403099) to MAB, an Undergraduate Research Fellowship from the SC Space Grant Consortium, and Clemson University Creative Inquiry.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Blazeck J, Hill A, Liu LQ, Knight R, Miller J, Pan A, Otoupal P, Alper HS (2014) Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production. Nat Commun 5. https://doi.org/10.1038/ncomms4131
- Bouatra S, Aziat F, Mandal R, Guo AC, Wilson MR, Knox C, Bjorndahl TC, Krishnamurthy R, Saleem F, Liu P, Dame ZT, Poelzer J, Huynh J, Yallou FS, Psychogios N, Dong E, Bogumil R, Roehring C, Wishart DS (2013) The human urine metabolome. PLoS One 8(9):e73076. https://doi.org/10.1371/journal.pone.0073076 CrossRefPubMedPubMedCentralGoogle Scholar
- Curran KA, Crook NC, Karim AS, Gupta A, Wagman AM, Alper HS (2014) Design of synthetic yeast promoters via tuning of nucleosome architecture. Nat Commun 5. https://doi.org/10.1038/ncomms5002
- Darvishi F, Nahvi I, Zarkesh-Esfahani H, Momenbeik F (2009) Effect of plant oils upon lipase and citric acid production in Yarrowia lipolytica yeast. J Biomed BiotechnolGoogle Scholar
- Evans CT, Ratledge C (1984) Effect of nitrogen-source on lipid-accumulation in oleaginous yeasts. J Gen Microbiol 130(Jul):1693–1704Google Scholar
- Goncalves FAG, Colen G, Takahashi JA (2014) Yarrowia lipolytica and its multiple applications in the biotechnological industry. Sci World JGoogle Scholar
- Haddouche R, Delessert S, Sabirova J, Neuveglise C, Poirier Y, Nicaud JM (2010) Roles of multiple acyl-CoA oxidases in the routing of carbon flow towards beta-oxidation and polyhydroxyalkanoate biosynthesis in Yarrowia lipolytica. FEMS Yeast Res 10(7):917–927. https://doi.org/10.1111/j.1567-1364.2010.00670.x CrossRefPubMedGoogle Scholar
- Imandi SB, Bandaru VVR, Somalanka SR, Bandaru SR, Garapati HR (2008) Application of statistical experimental designs for the optimization of medium constituents for the production of citric acid from pineapple waste. Bioresour Technol 99(10):4445–4450. https://doi.org/10.1016/j.biortech.2007.08.071 CrossRefPubMedGoogle Scholar
- Menezes AA, Cumbers J, Hogan JA, Arkin AP (2015) Towards synthetic biological approaches to resource utilization on space missions. J R Soc Interface 12(102)Google Scholar
- Rigouin C, Gueroult M, Croux C, Dubois G, Borsenberger V, Barbe S, Marty A, Daboussi F, Andre I, Bordes F (2017) Production of medium chain fatty acids by Yarrowia lipolytica: combining molecular design and TALEN to engineer the fatty acid synthase. ACS Synth Biol 6(10):1870–1879. https://doi.org/10.1021/acssynbio.7b00034 CrossRefPubMedGoogle Scholar
- Shaw AJ, Lam FH, Hamilton M, Consiglio A, MacEwen K, Brevnova EE, Greenhagen E, LaTouf WG, South CR, van Dijken H, Stephanopoulos G (2016) Metabolic engineering of microbial competitive advantage for industrial fermentation processes. Science 353(6299):583–586. https://doi.org/10.1126/science.aaf6159 CrossRefPubMedGoogle Scholar
- Wang GK, Xiong XC, Ghogare R, Wang PD, Meng YH, Chen SL (2016) Exploring fatty alcohol-producing capability of Yarrowia lipolytica. Biotechnol Biofuels 9Google Scholar
- Xue Z, Sharpe PL, Hong SP, Yadav NS, Xie D, Short DR, Damude HG, Rupert RA, Seip JE, Wang J, Pollak DW, Bostick MW, Bosak MD, Macool DJ, Hollerbach DH, Zhang H, Arcilla DM, Bledsoe SA, Croker K, McCord EF, Tyreus BD, Jackson EN, Zhu Q (2013) Production of omega-3 eicosapentaenoic acid by metabolic engineering of Yarrowia lipolytica. Nat Biotechnol 31(8):734–740. https://doi.org/10.1038/nbt.2622 CrossRefPubMedGoogle Scholar
- Zhao MX, Chi Z, Chi ZM, Madzak C (2013) The simultaneous production of single-cell protein and a recombinant antibacterial peptide by expression of an antibacterial peptide gene in Yarrowia lipolytica. Process Biochem 48(2):212–217. https://doi.org/10.1016/j.procbio.2013.01.003 CrossRefGoogle Scholar