Characterizing the Cellular Envelope and Secreted Compounds of Anaerobic Gut Fungi
EMSL Project ID
48839
Abstract
This proposal seeks use of EMSL resources to characterize the cellular envelope and secreted metabolites of anaerobic gut fungi. We have previously isolated three gut fungal strains representing different fungal genera (Piromyces sp finn, Neocallimastix sp G1, and Anaeromyces sp S4), which are powerful degraders of unpretreated biomass. Previous transcriptomic analysis of these strains (in collaboration with the JGI) suggest a reservoir of membrane-bound proteins that likely play a role in reprogramming fungal metabolism, as well as the existence of dozens of polyketide synthases within each strain. Given the minority role of gut fungi within their native microbial community, we hypothesize that they secrete polyketide defense compounds to distance microbial competitors. Further, we hypothesize that the membrane proteome, morphology, and composition of gut fungi dynamically evolve throughout the fungal life cycle, and as a function of available nutrients. Using three gut fungal strains, we will leverage existing transcriptomic data for proteomic validation, and connect this information to sub-cellular protein localization and metabolite secretion. Our project will specifically (1) establish the composition of the fungal cellular envelope by high-resolution spectroscopy and mass spectrometry. This information will lend insight into how gut fungi interact with their environment and how the surrounding environment and life cycle of the species can alter features of the cellular envelope. We will also (2) identify the polyketide compounds secreted by gut fungi via GC-MS and targeted NMR of extracellular secretions. Identified polyketides could find application as drop-in biofuels, and also serve as regulatory compounds that govern the dynamics of the microbial gut community. Outcomes of this project synergize with ongoing research efforts funded by DOE-BER, and in addition to polyketide biofuels could provide novel sugar transporters and receptors valuable to microbial engineering efforts. Collectively, this information will establish the molecular framework for anaerobic fungal hydrolysis in a dynamic microbial community, and will guide in the development of lignocellulosic biofuels.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2015-10-01
End Date
2018-03-31
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Henske JK, SE Wilken, KV Solomon, CR Smallwood, V Shutthanandan, JE Evans, MK Theodorou, and MA O'Malley. 2018. "Metabolic characterization of anaerobic fungi provides a path forward for bioprocessing of crude lignocellulose." Biotechnology and Bioengineering 115(4):874-884. doi:10.1002/bit.26515
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
Seppälä S ,Solomon K V,Gilmore S P,Henske J K,O'Malley M A 2016. "Mapping the Membrane Proteome of Anaerobic Gut Fungi Identifies a Wealth of Carbohydrate Binding Proteins and Transporters" Microbial Cell Factories 15():212. 10.1186/s12934-016-0611-7
Solomon KV, C Haitjema, JK Henske, SP Gilmore, D Borges-Rivera, A Lipzen, HM Brewer, SO Purvine, AT Wright, MK Theodorou, IV Grigoriev, A Regev, D Thompson, and MA O'Malley. 2016. "Early-branching Gut Fungi Possess A Large, And Comprehensive Array Of Biomass-Degrading Enzymes." Science 351(6278):1192-1195. doi:10.1126/science.aad1431
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