Effect of enzyme supplementation at moderate cellulase loadings on initial glucose and xylose release from corn stover solids pretreated by leading technologies.

Moderate loadings of cellulase enzyme supplemented with beta-glucosidase were applied to solids produced by ammonia fiber expansion (AFEX), ammonia recycle (ARP), controlled pH, dilute sulfuric acid, lime, and sulfur dioxide pretreatments to better understand factors that control glucose and xylose release following 24, 48, and 72 h of hydrolysis and define promising routes to reducing enzyme demands. Glucose removal was higher from all pretreatments than from Avicel cellulose at lower enzyme loadings, but sugar release was a bit lower for solids prepared by dilute sulfuric acid in the Sunds system and by controlled pH pretreatment than from Avicel at higher protein loadings. Inhibition by cellobiose was observed to depend on the type of substrate and pretreatment and hydrolysis times, with a corresponding impact of beta-glucosidase supplementation. Furthermore, for the first time, xylobiose and higher xylooligomers were shown to inhibit enzymatic hydrolysis of pure glucan, pure xylan, and pretreated corn stover, and xylose, xylobiose, and xylotriose were shown to have progressively greater effects on hydrolysis rates. Consistent with this, addition of xylanase and beta-xylosidase improved performance significantly. For a combined mass loading of cellulase and beta-glucosidase of 16.1 mg/g original glucan (about 7.5 FPU/g), glucose release from pretreated solids ranged from 50% to75% of the theoretical maximum and was greater for all pretreatments at all protein loadings compared to pure Avicel cellulose except for solids from controlled pH pretreatment and from dilute acid pretreatment by the Sunds pilot unit. The fraction of xylose released from pretreated solids was always less than for glucose, with the upper limit being about 60% of the maximum for ARP and the Sunds dilute acid pretreatments at a very high protein mass loading of 116 mg/g glucan (about 60 FPU).

Refining sweet sorghum to ethanol and sugar: economic trade-offs in the context of North China.

Reducing the use of non-renewable fossil energy reserves together with improving the environment are two important reasons that drive interest in the use of bioethanol as an automotive fuel. Conversion of sugar and starch to ethanol has been proven at an industrial scale in Brazil and the United States, respectively, and this alcohol has been able to compete with conventional gasoline due to various incentives. In this paper, we examined making ethanol from the sugar extracted from the juice of sweet sorghum and/or from the hemicellulose and cellulose in the residual sorghum bagasse versus selling the sugar from the juice or burning the bagasse to make electricity in four scenarios in the context of North China. In general terms, the production of ethanol from the hemicellulose and cellulose in bagasse was more favorable than burning it to make power, but the relative merits of making ethanol or sugar from the juice was very sensitive to the price of sugar in China. This result was confirmed by both process economics and analysis of opportunity costs. Thus, a flexible plant capable of making both sugar and fuel-ethanol from the juice is recommended. Overall, ethanol production from sorghum bagasse appears very favorable, but other agricultural residues such as corn stover and rice hulls would likely provide a more attractive feedstock for making ethanol in the medium and long term due to their extensive availability in North China and their independence from other markets. Furthermore, the process for residue conversion was based on particular design assumptions, and other technologies could enhance competitiveness while considerations such as perceived risk could impede applications.

Heat transfer considerations in design of a batch tube reactor for biomass hydrolysis.

Biologic conversion of inexpensive and abundant sources of cellulosic biomass offers a low-cost route to production of fuels and commodity chemicals that can provide unparalleled environmental, economic, and strategic benefits. However, low-cost, high-yield technologies are needed to recover sugars from the hemicellulose fraction of biomass and to prepare the remaining cellulose fraction for subsequent hydrolysis. Uncatalyzed hemicellulose hydrolysis in flow-through systems offers a number of important advantages for removal of hemicellulose sugars, and it is believed that oligomers could play an important role in explaining why the performance of flow-through systems differs from uncatalyzed steam explosion approaches. Thus, an effort is under way to study oligomer formation kinetics, and a small batch reactor is being applied to capture these important intermediates in a closed system that facilitates material balance closure for varying reaction conditions. In this article, heat transfer for batch tubes is analyzed to derive temperature profiles for different tube diameters and assess the impact on xylan conversion. It was found that the tube diameter must be <0.5 in. for xylan hydrolysis to follow the kinetics expected for a uniform temperature system at typical operating conditions.

Twenty years of trials, tribulations, and research progress in bioethanol technology: selected key events along the way.

The projected cost of ethanol production from cellulosic biomass has been reduced by almost a factor of four over the last 20 yr. Thus, it is now competitive for blending with gasoline, and several companies are working to build the first plants. However, technology development faced challenges at all levels. Because the benefits of bioethanol were not well understood, it was imperative to clarify and differentiate its attributes. Process engineering was invaluable in focusing on promising opportunities for improvements, particularly in light of budget reductions, and in tracking progress toward a competitive goal. Now it is vital for one or more commercial projects to be successful, and improving our understanding of process fundamentals will reduce the time and costs for commercialization. Additionally, the cost of bioethanol must be cut further to be competitive as a pure fuel in the open market, and aggressive technology advances are required to meet this target.

Cellulose and hemicellulose hydrolysis models for application to current and novel pretreatment processes.

Acids catalyze the hydrolysis of cellulose and hemicellulose to produce sugars that organisms can ferment to ethanol and other products. However, advanced low- and no-acid technologies are critical if we are to reduce bio-ethanol costs to be competitive as a pure fuel. We believe carbohydrate oligomers play a key role in explaining the performance of such hydrolysis processes and that kinetic models would help us understand their role. Various investigations have developed reaction rate expressions based on an Arrhenius temperature dependence that is first order in substrate concentration and close to first order in acid concentration. In this article, we evaluate these existing hydrolysis models with the goal of providing a foundation for a unified model that can predict performance of both current and novel pretreatment process configurations.

Characterization of recombinant E. coli ATCC 11303 (pLOI 297) in the conversion of cellulose and xylose to ethanol.

This work describes the characterization of recombinant Escherichia coli ATCC 11303 (pLOI 297) in the production of ethanol from cellulose and xylose. We have examined the fermentation of glucose and xylose, both individually and in mixtures, and the selectivity of ethanol production under various conditions of operation. Xylose metabolism was strongly inhibited by the presence of glucose. Ethanol was a strong inhibitor of both glucose and xylose fermentations; the maximum ethanol levels achieved at 37 degrees C and 42 degrees C were about 50 g/l and 25 g/l respectively. Simultaneous saccharification and fermentation of cellulose with recombinant E. coli and exogenous cellulose showed a high ethanol yield (84% of theoretical) in the hydrolysis regime of pH 5.0 and 37 degrees C. The selectivity of organic acid formation relative to that of ethanol increased at extreme levels of initial glucose concentration; production of succinic and acetic acids increased at low levels of glucose (< 1 g/l), and lactic acid production increased when initial glucose was higher than 100 g/l.

Study of the enzymatic hydrolysis of cellulose for production of fuel ethanol by the simultaneous saccharification and fermentation process.

The biochemical conversion of cellulosic biomass to ethanol, a promising alternative fuel, can be carried out efficiently and economically using the simultaneous saccharification and fermentation (SSF) process. The SSF integrates the enzymatic hydrolysis of cellulose to glucose, catalyzed by the synergistic action of cellulase and beta-glucosidase, with the fermentative synthesis of ethanol. Because the enzymatic step determines the ethanol. Because the enzymatic step determines the availability of glucose to the ethanologenic fermentation, the kinetic of cellulose hydrolysis by cellulase and beta-glucosidase and the susceptibility of the two enzymes to inhibition by hydrolysis and fermentation products are of significant importance to the SSF performance and were investigated under realistic SSF conditions. A previously developed SSF mathematical model was used to conceptualize the depolymerization of cellulose. The model was regressed to the collected data to determine the values of the enzyme parameters and was found to satisfactorily predict the kinetics of cellulose hydrolysis. Cellobiose and glucose were identified as the strongest inhibitors of cellulase and beta-glucosidase, respectively. Experimental and modeling results are presented in light of the impact of enzymatic hydrolysis on fuel ethanol production.

Fuel ethanol from cellulosic biomass.

Ethanol produced from cellulosic biomass is examined as a large-scale transportation fuel. Desirable features include ethanol's fuel properties as well as benefits with respect to urban air quality, global climate change, balance of trade, and energy security. Energy balance, feedstock supply, and environmental impact considerations are not seen as significant barriers to the widespread use of fuel ethanol derived from cellulosic biomass. Conversion economics is the key obstacle to be overcome. In light of past progress and future prospects for research-driven improvements, a cost-competitive process appears possible in a decade.

Evaluation of thermotolerant yeasts in controlled simultaneous saccharifications and fermentations of cellulose to ethanol.

Simultaneous saccharification and fermentation (SSF) experiments were performed at selected temperatures (37, 41, and 43 degrees C) to obtain comprehensive material balance and performance data for several promising strains of thermotolerant yeast. Parameters measured were ethanol concentration, yeast cell density, and residual sugar and cellulose concentrations. The three yeasts Saccharomyces uvarum, Candida brassicae, and C. lusitaniae and two mixed cultures of Brettanomyces clausenii with S. cerevisiae (mixed culture I) and C. Iusitaniae with S. uvarum (mixed culture II) exhibited rapid rates of fermentation, high ethanol yields, strong viability, or high cellobiase activity. Overall, mixed culture II at 41 degrees C performed better than either component yeast by themselves because it combined a cellobiose fermenting capability with the high ethanol tolerance and rapid glucose fermentation of conventional industrial yeasts. Thus, the mixed cultures provide good initial rates by preventing buildup of cellobiose (a strong inhibitor of enzyme activity) while attaining high ultimate yields of ethanol for high cellulase concentrations. However, C. brassicae and S. uvarum gave similar results to mixed culture II at 37 degrees C.