Metabolic engineering of the oleaginous yeast Rhodosporidium toruloides IFO0880 for lipid overproduction during high-density fermentation.
Natural lipids can be used to make biodiesel and many other value-added compounds. In this work, we explored a number of different metabolic engineering strategies for increasing lipid production in the oleaginous yeast Rhodosporidium toruloides IFO0880. These included increasing the expression of enzymes involved in different aspects of lipid biosynthesis-malic enzyme (ME), pyruvate carboxylase (PYC1), glycerol-3-P dehydrogenase (GPD), and stearoyl-CoA desaturase (SCD)-and deleting the gene PEX10, required for peroxisome biogenesis. Only malic enzyme and stearoyl-CoA desaturase, when overexpressed, were found to significantly increase lipid production. Only stearoyl-CoA desaturase, when overexpressed, further increased lipid production in a strain previously engineered to overexpress acetyl-CoA carboxylase (ACC1) and diacylglycerol acyltransferase (DGA1). Our best strain produced 27.4 g/L lipid with an average productivity of 0.31 g/L/h during batch growth on glucose and 89.4 g/L lipid with an average productivity of 0.61 g/L/h during fed-batch growth on glucose. These results further establish R. toruloides as a platform organism for the production of lipids and potentially other lipid-derived compounds from sugars.
Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, 600 S. Mathews Ave, Urbana, IL, 61810, USA.
Shuyan
S
Zhang
Department of Bioengineering, University of California, Berkeley, CA, USA.|Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Masakazu
M
Ito
Department of Bioengineering, University of California, Berkeley, CA, USA.|Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Jeffrey M
JM
Skerker
Department of Bioengineering, University of California, Berkeley, CA, USA.|Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Adam P
AP
Arkin
Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, 600 S. Mathews Ave, Urbana, IL, 61810, USA. cvrao@illinois.edu.
Christopher V
CV
Rao
D004798:Enzymes / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D017353:Gene Deletion; D015870:Gene Expression; D050356:Lipid Metabolism; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways; D012248:Rhodotorula / Q000235:genetics* / Q000378:metabolism*
D004798:Enzymes
Lipids; Metabolic engineering; Oleaginous yeast; Rhodosporidium toruloides