title: Ionic Liquids Impact the Bioenergy Feedstock-Degrading Microbiome and Transcription of Enzymes Relevant to Polysaccharide Hydrolysis.
abstract: Ionic liquid (IL) pretreatment is a promising approach for the conversion of lignocellulose to biofuels. The toxicity of residual IL, however, negatively impacts the performance of industrial enzymes and microorganisms in hydrolysis and fermentation. In this study, a thermophilic microbial community was cultured on switchgrass amended with various levels of the ionic liquid 1-ethyl-3-methylimidazolium acetate. Changes in the microbial community composition and transcription of genes relevant to IL tolerance and lignocellulose hydrolysis were quantified. Increasing the level of IL to 0.1% (wt) led to increased levels of relative abundance and transcription in organisms of the phylumFirmicutes. Interestingly, IL concentrations of up to 1% (wt) also resulted in greater xylanase transcription and enzyme activity as well as increased transcription of endoglucanase, beta-glucosidase, and IL tolerance genes compared to communities without IL. IL levels above 1% (wt) resulted in decreased enzyme activity and transcription of genes involved in lignocellulose hydrolysis. The results indicate that moderate levels of IL select for thermophilic microorganisms that not only tolerate IL but also effectively hydrolyze lignocellulose from switchgrass. Discovery of IL-tolerant organisms and enzymes is critical for the development of biological processes that convert IL-pretreated biomass to biofuels and chemicals. Employing metatranscriptomic analysis of enrichment cultures can facilitate the discovery of microorganisms and enzymes that may be active in the presence of toxic compounds such as ionic liquids.IMPORTANCEPretreatment using ionic liquids (IL) is a promising approach for the conversion of lignocellulose to biofuels. Because IL can be inhibitory to enzymes and microorganisms involved in downstream hydrolysis and fermentation steps, discovery of IL-tolerant organisms and enzymes is critical for advancing this technology. Employing metatranscriptomics in the analysis of IL-enriched cultures facilitated tracking of dynamic changes in a complex microbial community at the level of gene transcription and doing so with genome resolution. Specific organisms were discovered that could simultaneously tolerate a moderate IL concentration and transcribe a diverse array of cellulolytic enzymes. Gene sequences of cellulolytic enzymes and efflux pumps from those same organisms were also identified, providing important resources for future research on engineering IL-tolerant organisms and enzymes.
authors.affiliation: Joint BioEnergy Institute, Emeryville, California, USA; Graduate Institute of Biomedical Informatics, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan.
authors.forename: Yu-Wei
authors.initials: YW
authors.lastname: Wu
authors.affiliation: Biological and Agricultural Engineering, University of California, Davis, Davis, California, USA; Biosystems Engineering, Auburn University, Auburn, Alabama, USA.
authors.forename: Brendan
authors.identifier: 0000-0001-6854-9612
authors.initials: B
authors.lastname: Higgins
authors.affiliation: Biological and Agricultural Engineering, University of California, Davis, Davis, California, USA.
authors.forename: Chaowei
authors.initials: C
authors.lastname: Yu
authors.affiliation: Biological and Agricultural Engineering, University of California, Davis, Davis, California, USA.
authors.forename: Amitha P
authors.initials: AP
authors.lastname: Reddy
authors.affiliation: Biological and Agricultural Engineering, University of California, Davis, Davis, California, USA.
authors.forename: Shannon
authors.initials: S
authors.lastname: Ceballos
authors.affiliation: Biological and Agricultural Engineering, University of California, Davis, Davis, California, USA.
authors.forename: Lawrence D
authors.initials: LD
authors.lastname: Joh
authors.affiliation: Joint BioEnergy Institute, Emeryville, California, USA; Biological and Materials Science Center, Sandia National Laboratories, Livermore, California, USA.
authors.forename: Blake A
authors.initials: BA
authors.lastname: Simmons
authors.affiliation: Joint BioEnergy Institute, Emeryville, California, USA; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
authors.forename: Steven W
authors.initials: SW
authors.lastname: Singer
authors.affiliation: Joint BioEnergy Institute, Emeryville, California, USA; Biological and Agricultural Engineering, University of California, Davis, Davis, California, USA.
authors.forename: Jean S
authors.identifier: 0000-0002-1455-8254
authors.initials: JS
authors.lastname: VanderGheynst
keywords: 1-ethyl-3-methylimidazolium acetate; cellulase; hemicellulase; ionic liquid