Rewiring Yarrowia Lipolytica for Production of Secreted, Fuel-like Alkane Products
Abstract
Advances in metabolic engineering and synthetic biology enable a newfound control of microbial metabolism. Despite this fact, we still rely heavily on petroleum to source most of the feedstocks used in the chemical industry and fuels arena. Yet, it is well established that alternative molecules such as mid to longer-chain hydrocarbons would make better fuels due to more favorable energy density, stability, and corrosion properties. Moreover, establishing these products from waste products in an organism capable of “on demand” production would provide a step-forward for interests in the Navy and other Department of Defense initiatives. Thus, the goal of this work is to establish the synthetic biology capacity and strain engineering to create a cellular platform for the production of secreted fuel-like saturated alkanes around 16 carbons in chain length. This work builds from prior success in our lab in which we were able to rewire the oleaginous yeast Yarrowia lipolytica to produce upwards of 90% of dry cell weight lipid and up to 40 g/L of production in small-scale bioreactors. The objectives of this research of this work will comprise: (1) complementing Y. lipolytica with novel, synthetic pathways for shorter chain fatty acids and alkanes, (2) establishing a strategy for evolving exporter proteins, (3) creating a flexible platform for secreted fuel-like alkanes in Y. lipolytica, and (4) establishing novel synthetic biology approaches broadly applicable to additional nonconventional hosts. The approaches in this proposal will enable the high level production of many new oleochemicals as will be demonstrated through straight-chain alkanes In accomplishing these objectives, we will rewire metabolites in cells to produce alkanes, refactor the fatty acid synthase complex to enable efficient production of key starting metabolites, perform directed evolution on exporter proteins to secrete alkanes, and develop critical synthetic biology capacities to rapidly engineering this host (as well as other nonconventional fungal organisms. Through these efforts, we will work toward the long-term goal of engineering an organism for on-site fuel production from waste streams as this host organism is uniquely capable of using chemical and sewage waste streams as carbon sources. These improvements and research tasks support ONR and Defense Missions under the directorate of Division 341 and 342 Research goals aimed at using metabolic engineering and synthetic biology to improve bioconversion capacity. In general, novel paradigms for improving cellular bioconversion capacity and cellular phenotypes will enable a prolific use of this platform for producing desirable chemical and precursors of interest for the Navy and other Defense missions.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Aug 12, 2016
- Source ID
- N000141512785
Entities
People
- Hal S. Alper
Organizations
- Office of Naval Research
- United States Navy
- University of Texas at Austin