The proteome and metabolome of OSER, cellular factories promoting high level terpene synthesis in filamentous fungi.
EMSL Project ID
49850
Abstract
Terpene compounds are a promising source of renewable biofuels that have many favorable characteristics such as high energy content and compatibility with existing energy-use technologies. While most attention has been paid to terpenes produced by plants, filamentous fungi also can naturally synthesize high levels of these compounds. However, there is a lack of knowledge of the cellular location of terpenoid biosynthetic pathways, the channeling of precursor molecules and intermediates, and the coordination of primary metabolism and secondary terpenoid biosynthetic pathways that result in maximum terpenoid synthesis. Sesquiterpene compounds called trichothecenes are conditionally expressed in the fungus Fusarium graminearum. Recently we have discovered that induction of trichothecene biosynthesis is linked to profound changes in the structure the fungal endomembrane network. Several trichothecene biosynthetic enzymes as well as HMG CoA reductase, the enzyme catalyzing synthesis of terpene precursor mevalonate, localize to Organized Smooth Endoplasmic Reticulum (OSER). This proposal seeks to further characterize the structure and content of these cellular terpenoid factories with the ultimate goal of using that information for improved terpene engineering and output. Specifically we propose to isolate fluorescently tagged OSER by flow cytometry and determine their protein and terpene content utilizing analytical resources at EMSL. EMSL FACS, proteomics and metabolomics facilities and expertise are essential for the success of the project. The OSER proteome data will be used to determine whether all known F. graminearum trichothecene biosynthetic enzymes are located in these organelles and to detect the presence of other unknown proteins that may be important for novel terpenoid synthesis or OSER biogenesis. Metabolomic data will be used to confirm OSER as the site of trichothecene biosynthesis, to detect potential pathway intermediates and perhaps to identify novel terpenes produce therein. Knowledge gained by this research will be used to guide functional studies aimed at expression of heterologous terpenoids, maximizing terpene expression, and engineering terpene pathways in filamentous fungi.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2017-10-01
End Date
2020-03-31
Status
Closed
Released Data Link
Team
Principal Investigator