title: Heterologous production of pentane in the oleaginous yeast Yarrowia lipolytica.
abstract: The complete biosynthetic replacement of petroleum transportation fuels requires a metabolic pathway capable of producing short chain n-alkanes. Here, we report and characterize a proof-of-concept pathway that enables microbial production of the C5 n-alkane, pentane. This pathway utilizes a soybean lipoxygenase enzyme to cleave linoleic acid to pentane and a tridecadienoic acid byproduct. Initial expression of the soybean lipoxygenase enzyme within a Yarrowia lipolytica host yielded 1.56 mg/L pentane. Efforts to improve pentane yield by increasing substrate availability and strongly overexpressing the lipoxygenase enzyme successfully increased pentane production three-fold to 4.98 mg/L. This work represents the first-ever microbial production of pentane and demonstrates that short chain n-alkane synthesis is conceivable in model cellular hosts. In this regard, we demonstrate the potential pliability of Y. lipolytica toward the biosynthetic production of value-added molecules from its generous fatty acid reserves.
authors.affiliation: McKetta Department of Chemical Engineering, The University of Texas at Austin, 200 E Dean Keeton Street Stop C0400, Austin, TX 78712, USA.
authors.forename: John
authors.initials: J
authors.lastname: Blazeck
authors.forename: Leqian
authors.initials: L
authors.lastname: Liu
authors.forename: Rebecca
authors.initials: R
authors.lastname: Knight
authors.forename: Hal S
authors.initials: HS
authors.lastname: Alper
mesh_terms: D000446:Aldehyde-Lyases / Q000235:genetics / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D008084:Lipoxygenase / Q000235:genetics / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D010084:Oxidation-Reduction; D010420:Pentanes / Q000032:analysis / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D030262:Soybean Proteins / Q000235:genetics / Q000378:metabolism; D013025:Glycine max / Q000201:enzymology / Q000235:genetics; D025062:Yarrowia / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*
chemical_list: D003470:Culture Media; D010420:Pentanes; D030262:Soybean Proteins; C033353:pentane; D003577:Cytochrome P-450 Enzyme System; D008084:Lipoxygenase; D000446:Aldehyde-Lyases; C054540:hydroperoxide lyase