Comprehensive Characterization of the Cellulosomes from Anaerobic Gut Fungi
EMSL Project ID
49765
Abstract
This proposal seeks use of EMSL resources to characterize the secreted, multi-enzyme cellulosomes that originate from anaerobic gut fungi. Cellulosomes enable anaerobic fungi to tether different types of biomass-degrading enzymes together for synergistic breakdown & conversion of plant biomass. However, cellulosome structure, sub-cellular localization & range of activities remain unexplored. We have previously isolated three gut fungal strains representing different fungal genera (Piromyces sp finn, Neocallimastix sp G1, and Anaeromyces sp S4), which secrete cellulosomes that are powerful degraders of unpretreated biomass. Previous genomic, transcriptomic, and biochemical analysis of these strains (in collaboration with the JGI & EMSL) have revealed the "parts list" that comprise cellulosomes in fungi, and suggest a unique scaffolding mechanism, localization, and post-translational modifications inherent in these complexes that could govern their functional rearrangement. We seek to partner with EMSL to (1) develop targeted labeling approaches, imaging, & proteomics to reveal how cellulosomes are attached to fungi, as well how their localization changes as a function of their environment. Further, we will (2) characterize the post-translational modifications and putative kinase/phosphatase rearrangement mechanisms within cellulosomes via targeted proteomics. Finally, we will combine advanced imaging and mass spectrometry to (3) determine how biomass substrates influence secretion of cellulosomes relative to free enzymes. Outcomes of this project synergize with ongoing research efforts funded by DOE-BER, and will reveal critical attributes of fungal cellulosomes that can be engineered and exploited for bio-based fuel & chemical production.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2017-10-01
End Date
2021-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Related Publications
Henske J.K., S.P. Gilmore, D. Knop, F.J. Cunningham, J.A. Sexton, C.R. Smallwood, and V. Shutthanandan, et al. 2017. "Transcriptomic characterization of Caecomyces churrovis: a novel, non-rhizoid-forming lignocellulolytic anaerobic fungus." Biotechnology for Biofuels 10. PNNL-SA-134108. doi:10.1186/s13068-017-0997-4
Wilken S., S. Seppala, T.S. Lankiewicz, M. Saxena, J.K. Henske, A. Salamov, and I.V. Grigoriev, et al. 2020. "Genomic and Proteomic Biases Inform Metabolic Engineering Strategies for Anaerobic Fungi." Metabolic Engineering Communications 10. doi:10.1016/j.mec.2019.e00107