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Henske, John K. ; Gilmore, Sean P. ; Haitjema, Charles H. ; Solomon, Kevin V. ; O'Malley, Michelle A. ( , AIChE Journal)
Anaerobic fungi are among the most active plant‐degrading microbes in nature. Increased insight into the mechanisms and environmental cues that regulate fungal hydrolysis would better inform bioprocessing strategies to depolymerize lignocellulose. Here, we compare the response of three strains of anaerobic fungi (
Piromyces finnis ,Anaeromyces robustus , andNeocallimastix californiae ) to catabolite regulation by simple carbohydrates. Anaerobic fungi exhibited high enzymatic activity against crystalline cellulose, which was repressed upon incubation with free sugars. Cellulolytic degradation was also inhibited when fungi were exposed to sugars they did not metabolize, suggesting a general mode of catabolite repression. RNA‐Seq experiments in the presence of excess glucose confirmed repression of carbohydrate active enzymes during sugar uptake, and offer a path towards unmasking the function of co‐regulated genes that could be involved in biomass degradation. Overall, these results suggest that sugar‐rich hydrolysates tune the behavior of anaerobic fungi by dampening production of their biomass‐degrading enzymes. © 2018 American Institute of Chemical EngineersAIChE J , 64: 4263–4270, 2018 -
Henske, John K. ; Wilken, St. Elmo ; Solomon, Kevin V. ; Smallwood, Chuck R. ; Shutthanandan, Vaithiyalingam ; Evans, James E. ; Theodorou, Michael K. ; O'Malley, Michelle A. ( , Biotechnology and Bioengineering)
Abstract The conversion of lignocellulose‐rich biomass to bio‐based chemicals and higher order fuels remains a grand challenge, as single‐microbe approaches often cannot drive both deconstruction and chemical production steps. In contrast, consortia based bioprocessing leverages the strengths of different microbes to distribute metabolic loads and achieve process synergy, product diversity, and bolster yields. Here, we describe a biphasic fermentation scheme that combines the lignocellulolytic action of anaerobic fungi isolated from large herbivores with domesticated microbes for bioproduction. When grown in batch culture, anaerobic fungi release excess sugars from both cellulose and crude biomass due to a wealth of highly expressed carbohydrate active enzymes (CAZymes), converting as much as 49% of cellulose to free glucose. This sugar‐rich hydrolysate readily supports growth of
Saccharomyces cerevisiae , which can be engineered to produce a range of value‐added chemicals. Further, construction of metabolic pathways from transcriptomic data reveals that anaerobic fungi do not catabolize all sugars that their enzymes hydrolyze from biomass, leaving other carbohydrates such as galactose, arabinose, and mannose available as nutritional links to other microbes in their consortium. Although basal expression of CAZymes in anaerobic fungi is high, it is drastically amplified by cellobiose breakout products encountered during biomass hydrolysis. Overall, these results suggest that anaerobic fungi provide a nutritional benefit to the rumen microbiome, which can be harnessed to design synthetic microbial communities that compartmentalize biomass degradation and bioproduct formation.