Deconstruction

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GLBRC's Deconstruction Research Area

Deconstruction

GLBRC's Deconstruction Lead takes top academic slot

Located at the intersection of the U.S.’s agricultural heartland and its northern forests, the GLBRC has access to a rich diversity of raw biomass for study. The Center's Deconstruction research focuses on identifying the best combinations of enzymes, chemicals, and physical processing methods for enhancing the digestibility of specific biomass sources.

Learn about the Center's research approach

Deconstruction Leadership

Deconstruction Lead

Dale is an expert on making ethanol from cellulose, plant stalks, grass, corn cobs and other woody plant parts and has developed a patented process called ammonia fiber expansion (AFEXTM), which makes the breakdown of cellulose more efficient, thus tackling...

Deconstruction Lead

Fox's research goals are to define the structure and the reactivity of the active site diiron center, to probe the catalytic contributions of the active site protein residues and to determine the consequences of protein-protein and protein-substrate interactions on the...

Project Overview

A biofuels reactor designed to produce ethanol at Michigan State University's Biomass Conversion Research Lab (BCRL)GLBRC Deconstruction research maintains a focus on the entire biofuels production pipeline: in addition to identifying and improving natural cellulose-degrading enzymes extracted from diverse environments, researchers apply unique biomass pretreatment technologies—such as ammonia fiber expansion (AFEX™), alkaline hydrogen peroxide (AHP), and extractive ammonia (EA)—that enable conversion technologies to maximize plant biomass utilization.. Researchers also explore strategies to add value to these processes by developing co-products from materials that would otherwise be treated as waste, such as lignin. Specific deconstruction projects include:

  • Pretreatment effects on biomass, alkaline peroxide pretreatment, fuel production from alkaline-pretreated biomass
  • Optimization of enzymes for biomass conversion, discovery of natural cellulolytic microbes, identification of novel microbial enzymes, and combinatorial discovery of enzymes and proteins

Deconstruction Publications

Next-generation ammonia pretreatment enhances cellulosic biofuel production

Leonardo da Costa Sousa; Mingjie Jin; Shishir P.S. Chundawat; Vijay Bokade; Xiaoyu Tang; Ali Azarpira; Fachuang Lu; Utku Avci; James Humpula; Nirmal Uppugundla; Christa Gunawan; Sivakumar Pattathil; Albert M. Cheh; Ninad Kothari; Rajeev Kumar; John Ralph; Michael G. Hahn; Charles E. Wyman; Singh. Seema; Blake A. Simmons; Bruce E. Dale; Venkatesh Balan

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2016

Prediction of cell wall properties and response to deconstruction using alkaline pretreatment in diverse maize genotypes using Py-MBMS and NIR

Muyang Li; Daniel L. Williams; Marlies Heckwolf; Natalia de Leon; Shawn Kaeppler; Robert W. Sykes; David Hodge

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2016

Quantifying pretreatment degradation compounds in solution and accumulated by cells during solids and yeast recycling in the Rapid Bioconversion with Integrated recycling Technology process using AFEX™ corn stover

Cory Sarks; Alan Higbee; Jeff Piotrowski; Saisi Xue; Joshua J. Coon; Trey K. Sato; Mingjie Jin; Venkatesh Balan; Bruce E. Dale

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2016

Effects of degradation products (low molecular weight compounds produced during pretreatment) on the microbes used in the RaBIT (Rapid Bioconversion with Integrated recycling Technology) process that reduces enzyme usage up to 40% by efficient enzyme recycling were studied. Chemical genomic profiling was performed, showing no yeast response differences in hydrolysates produced during RaBIT enzymatic hydrolysis. Concentrations of degradation products in solution were quantified after different enzymatic hydrolysis cycles and fermentation cycles. Intracellular degradation product concentrations were also measured following fermentation. Degradation product concentrations in hydrolysate did not change between RaBIT enzymatic hydrolysis cycles; the cell population retained its ability to oxidize/reduce (detoxify) aldehydes over five RaBIT fermentation cycles; and degradation products accumulated within or on the cells as RaBIT fermentation cycles increased. Synthetic hydrolysate was used to confirm that pretreatment degradation products are the sole cause of decreased xylose consumption during RaBIT fermentations.

Saccharification of newspaper waste after ammonia fiber expansion or extractive ammonia

Salvatore Montella; Venkatesh Balan; Leonardo da Costa Sousa; Christa Gunawan; Simona Giacobbe; Olimpia Pepe; Vincenza Faraco

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2016

The lignocellulosic fractions of municipal solid waste (MSW) can be used as renewable resources due to the widespread availability, predictable and low pricing and suitability for most conversion technologies. In particular, after the typical paper recycling loop, the newspaper waste (NW) could be further valorized as feedstock in biorefinering industry since it still contains up to 70 % polysaccharides. In this study, two different physicochemical methods—ammonia fiber expansion (AFEX) and extractive ammonia (EA) were tested for the pretraetment of NW. Furthermore, based on the previously demonstrated ability of the recombinant enzymes endocellulase rCelStrep, α-l-arabinofuranosidase rPoAbf and its evolved variant rPoAbf F435Y/Y446F to improve the saccharification of different lignocellulosic pretreated biomasses (such as corn stover and Arundo donax), in this study these enzymes were tested for the hydrolysis of pretreated NW, with the aim of valorizing the lignocellulosic fractions of the MSW. In particular, a mixture of purified enzymes containing cellulases, xylanases and accessory hemicellulases, was chosen as reference mix and rCelStrep and rPoAbf or its variant were replaced to EGI and Larb. The results showed that these enzymatic mixes are not suitable for the hydrolysis of NW after AFEX or EA pretreatment. On the other hand, when the enzymes rCelStrep, rPoAbf and rPoAbf F435Y/Y446F were tested for their effect in hydrolysis of pretreated NW by addition to a commercial enzyme mixture, it was shown that the total polysaccharides conversion yield reached 37.32 % for AFEX pretreated NW by adding rPoAbf to the mix whilst the maximum sugars conversion yield for EA pretreated NW was achieved 40.80 % by adding rCelStrep. The maximum glucan conversion yield obtained (45.61 % for EA pretreated NW by adding rCelStrep to the commercial mix) is higher than or comparable to those reported in recent manuscripts adopting hydrolysis conditions similar to those used in this study.

Saccharification of thermochemically pretreated cellulosic biomass using native and engineered cellulosomal enzyme systems

Shishir P.S. Chundawat; Chad D. Paavola; Babu Raman; Matthieu Nouailler; Suzanne L. Chan; Jonathan R. Mielenz; Veronique Receveur-Brechot; Jonathan D. Trent; Bruce E. Dale

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2016

Consolidated bioprocessing (CBP) of pretreated lignocellulosic biomass using microbes like Clostridium thermocellum allows simultaneous polysaccharide saccharification and sugar fermentation to produce fuels or chemicals using a one-pot process. C. thermocellum is a thermophilic bacterium that deconstructs biomass using large multi-enzyme complexes called cellulosomes. Characterization of cellulosomal enzymes tethered to native or engineered scaffoldin proteins has revealed that enzyme complexation is critical to the bacterium's cellulolytic ability. However, we have a limited understanding of the impact of enzyme complexation on the saccharification efficiency of various forms of industrially relevant pretreated biomass substrates. Here, we compared the hydrolytic activity of the most abundant cellulosomal enzymes from C. thermocellum and investigate the importance of enzyme complexation using a model engineered protein scaffold (called 'rosettasome'). The hydrolytic performance of non-complexed enzymes, enzyme-rosettasome (or rosettazyme) complexes, and cellulosomes was tested on distinct cellulose allomorphs formed during biomass pretreatment. The scaffold-immobilized enzymes always gave higher activity than free enzymes. However, cellulosomes exhibited higher activity than rosettazyme complexes. This was likely due to the greater flexibility of the native versus engineered scaffold, as deciphered using small angle X-ray scattering. Surprisingly, scaffold-tethered enzymes also gave comparable activity on all the cellulose allomorphs tested, which is unlike the preferential activity of non-complexed cellulases seen for certain allomorph forms. Tethered enzyme complexes also gave lower saccharification yields on industrially relevant lignin-rich switchgrass than cellulose alone. In summary, we find that the type of pretreatment can significantly impact the saccharification efficiency of cellulosomal enzymes for various CBP scenarios.

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