Yoon Y. Lee

A review on alkaline pretreatment technology for bioconversion of lignocellulosic biomass.

The native form of lignocellulosic biomass is resistant to enzymatic breakdown. A well-designed pretreatment that can promote enzymatic hydrolysis of biomass with reasonable processing cost is therefore necessary. To this end, a number of different types of pretreatment technologies have been developed with a common goal of making biomass more susceptible to enzymatic saccharification. Among those, a pretreatment method using alkaline reagent has emerged as one of the most viable process options due primarily to its strong pretreatment effect and relatively simple process scheme. The main features of alkaline pretreatment are that it selectively removes lignin without degrading carbohydrates, and increases porosity and surface area, thereby enhancing enzymatic hydrolysis. In this review, the leading alkaline pretreatment technologies are described and their features and comparative performances are discussed from a process viewpoint. Attempts were also made to give insights into the chemical and physical changes of biomass brought about by pretreatment.

Process and technoeconomic analysis of leading pretreatment technologies for lignocellulosic ethanol production using switchgrass

Six biomass pretreatment processes to convert switchgrass to fermentable sugars and ultimately to cellulosic ethanol are compared on a consistent basis in this technoeconomic analysis. The six pretreatment processes are ammonia fiber expansion (AFEX), dilute acid (DA), lime, liquid hot water (LHW), soaking in aqueous ammonia (SAA), and sulfur dioxide-impregnated steam explosion (SO(2)). Each pretreatment process is modeled in the framework of an existing biochemical design model so that systematic variations of process-related changes are consistently captured. The pretreatment area process design and simulation are based on the research data generated within the Biomass Refining Consortium for Applied Fundamentals and Innovation (CAFI) 3 project. Overall ethanol production, total capital investment, and minimum ethanol selling price (MESP) are reported along with selected sensitivity analysis. The results show limited differentiation between the projected economic performances of the pretreatment options, except for processes that exhibit significantly lower monomer sugar and resulting ethanol yields.

Pretreatment of corn stover by low-liquid ammonia recycle percolation process

A pretreatment method using aqueous ammonia was investigated with the intent of minimizing the liquid throughput. This process uses a flow-through packed column reactor (or percolation reactor). In comparison to the ammonia recycle percolation (ARP) process developed previously in our laboratory, this process significantly reduces the liquid throughput to one reactor void volume in packed bed (2.0–4.7 mL of liquid/g of corn stover) and, thus, is termed low-liquid ARP (LLARP). In addition to attaining short residence time and reduced energy input, this process achieves 59–70% of lignin removal and 48–57% of xylan retention. With optimum operation of the LLARP to corn stover, enzymatic digestibilities of 95, 90 and 86% were achieved with 60, 15, and 7.5 filter paper units/g of glucan, respectively. In the simultaneous saccharification and fermentation test of the LLARP samples using Saccharomyces cerevisiae (NREL-D5A), an ethanol yield of 84% of the theoretical maximum was achieved with 6% (w/v) glucan loading. In the simultaneous saccharification and cofermentation (SSCF) test using recombinant Escherichia coli (KO11), both the glucan and xylan in the solid were effectively utilized, giving an overall ethanol yield of 109% of the theoretical maximum based on glucan, a clear indication that the xylan content was converted into ethanol. The xylooligomers existing in the LLARP effluent were not effectively hydrolyzed by cellulase enzyme, achieving only 60% of digestibility. SSCF of the treated corn stover was severely hampered when the substrate was supplemented with the LLARP effluent, giving only 56% the overall yield of ethanol. The effluent appears to significantly inhibit cellulase and microbial activities.

Comparative data on effects of leading pretreatments and enzyme loadings and formulations on sugar yields from different switchgrass sources

Dilute sulfuric acid (DA), sulfur dioxide (SO(2)), liquid hot water (LHW), soaking in aqueous ammonia (SAA), ammonia fiber expansion (AFEX), and lime pretreatments were applied to Alamo, Dacotah, and Shawnee switchgrass. Application of the same analytical methods and material balance approaches facilitated meaningful comparisons of glucose and xylose yields from combined pretreatment and enzymatic hydrolysis. Use of a common supply of cellulase, beta-glucosidase, and xylanase also eased comparisons. All pretreatments enhanced sugar recovery from pretreatment and subsequent enzymatic hydrolysis substantially compared to untreated switchgrass. Adding beta-glucosidase was effective early in enzymatic hydrolysis while cellobiose levels were high but had limited effect on longer term yields at the enzyme loadings applied. Adding xylanase improved yields most for higher pH pretreatments where more xylan was left in the solids. Harvest time had more impact on performance than switchgrass variety, and microscopy showed changes in different features could impact performance by different pretreatments.

Ammonia Recycled Percolation Process for Pretreatment of Herbaceous Biomass

An ammonia-based pretreatment method termed ammonia recycled percolation (ARP) was developed for pretreating herbaceous biomass like corn cobs/stover mixture (CCSM) and switchgrass. The process involves treatment of biomass with aqueous ammonia through a percolation reactor (packed-bed, flow-through type). The effects temperature, reaction time, and ammonia concentration were studied. The extent of delignification in the ARP process was in the range of 60-80% for CCSM and 65-85% for switchgrass. The ARP process solubilized significant amounts of the hemicellulose fraction into the pretreatment effluent, yet left most of the glucan fraction intact. The experimental data on CCSM and switchgrass indicate that the ARP is a highly effective pretreatment method. Near-complete conversion of cellulose to glucose was obtained by enzymatic hydrolysis of ARP-treated solid samples of CCSM, whereas conversion was slightly lower for switchgrass. The rate of enzymatic hydrolysis of ARP-treated samples was substantially higher than that of a-cellulose. The ARP effluents were evaluated for fermentability/toxicity by the xylose-fermenting yeastPichia stipitis(NRRL Y-11545). The adaptability of ARP-treated solid samples to simultaneous saccharification and fermentation (SSF) was tested for ethanol production using cellulase enzyme and the yeast, Saccharomyces cerevisiae (NREL, D5A).

Comparative material balances around pretreatment technologies for the conversion of switchgrass to soluble sugars

For this project, six chemical pretreatments were compared for the Consortium for Applied Fundamentals and Innovation (CAFI): ammonia fiber expansion (AFEX), dilute sulfuric acid (DA), lime, liquid hot water (LHW), soaking in aqueous ammonia (SAA), and sulfur dioxide (SO(2)). For each pretreatment, a material balance was analyzed around the pretreatment, optional post-washing step, and enzymatic hydrolysis of Dacotah switchgrass. All pretreatments+enzymatic hydrolysis solubilized over two-thirds of the available glucan and xylan. Lime, post-washed LHW, and SO(2) achieved >83% total glucose yields. Lime, post-washed AFEX, and DA achieved >83% total xylose yields. Alkaline pretreatments, except AFEX, solubilized the most lignin and a portion of the xylan as xylo-oligomers. As pretreatment pH decreased, total solubilized xylan and released monomeric xylose increased. Low temperature-long time or high temperature-short time pretreatments are necessary for high glucose release from late-harvest Dacotah switchgrass but high temperatures may cause xylose degradation.

Study on the decreased sugar yield in enzymatic hydrolysis of cellulosic substrate at high solid loading.

Current technology for conversion of biomass to ethanol is an enzyme-based biochemical process. In bioethanol production, achieving high sugar yield at high solid loading in enzymatic hydrolysis step is important from both technical and economic viewpoints. Enzymatic hydrolysis of cellulosic substrates is affected by many parameters, including an unexplained behavior that the glucan digestibility of substrates by cellulase decreased under high solid loadings. A comprehensive study was conducted to investigate this phenomenon by using Spezyme CP and Avicel as model cellulase and cellulose substrate, respectively. The hydrolytic properties of the cellulase under different substrate concentrations at a fixed enzyme-to-substrate ratio were characterized. The results indicate that decreased sugar yield is neither due to the loss of enzyme activity at a high substrate concentration nor due to the higher end-product inhibition. The cellulase adsorption kinetics and isotherm studies indicated that a decline in the binding capacity of cellulase may explain the long-observed but little-understood phenomenon of a lower substrate digestibility with increased substrate concentration. The mechanism how the enzyme adsorption properties changed at high substrate concentration was also discussed in the context of exploring the improvement of the cellulase-binding capacity at high substrate loading.

Comparative study on enzymatic digestibility of switchgrass varieties and harvests processed by leading pretreatment technologies

Feedstock quality of switchgrass for biofuel production depends on many factors such as morphological types, geographic origins, maturity, environmental and cultivation parameters, and storage. We report variability in compositions and enzymatic digestion efficiencies for three cultivars of switchgrass (Alamo, Dacotah and Shawnee), grown and harvested at different locations and seasons. Saccharification yields of switchgrass processed by different pretreatment technologies (AFEX, dilute sulfuric acid, liquid hot water, lime, and soaking in aqueous ammonia) are compared in regards to switchgrass genotypes and harvest seasons. Despite its higher cellulose content per dry mass, Dacotah switchgrass harvested after wintering consistently gave a lower saccharification yield than the other two varieties harvested in the fall. The recalcitrance of upland cultivars and over-wintered switchgrass may require more severe pretreatment conditions. We discuss the key features of different pretreatment technologies and differences in switchgrass cultivars and harvest seasons on hydrolysis performance for the applied pretreatment methods.

Application of cellulase and hemicellulase to pure xylan, pure cellulose, and switchgrass solids from leading pretreatments

Accellerase 1000 cellulase, Spezyme CP cellulase, β-glucosidase, Multifect xylanase, and beta-xylosidase were evaluated for hydrolysis of pure cellulose, pure xylan, and switchgrass solids from leading pretreatments of dilute sulfuric acid, sulfur dioxide, liquid hot water, lime, soaking in aqueous ammonia, and ammonia fiber expansion. Distinctive sugar release patterns were observed from Avicel, phosphoric acid swollen cellulose (PASC), xylan, and pretreated switchgrass solids, with accumulation of significant amounts of xylooligomers during xylan hydrolysis. The strong inhibition of cellulose hydrolysis by xylooligomers could be partially attributed to the negative impact of xylooligomers on cellulase adsorption. The digestibility of pretreated switchgrass varied with pretreatment but could not be consistently correlated to xylan, lignin, or acetyl removal. Initial hydrolysis rates did correlate well with cellulase adsorption capacities for all pretreatments except lime, but more investigation is needed to relate this behavior to physical and compositional properties of pretreated switchgrass.

Fractionation of herbaceous biomass by ammonia-hydrogen peroxide percolation treatment

Treatment with ammonia and hydrogen peroxide was investigated as a means of fractionating herbaceous biomass. The main feature of this process is that aqueous forms of these reagents are pumped simultaneously into a packed-bed flow-through-type reactor (percolation reactor) under a semibatch mode with ammonia being recycled. Experimental tests on corn cobs/stover mixture (CCSM) and switchgrass feedstocks have proven that a high degree of fractionation of biomass into three major components is attainable under this process scheme. The extent of delignification was 94–99% It was achieved at a representative condition of 170°C, 0.28 g loading of H2O2/g biomass, and 10 wt% ammonia concentration. At the same time, about 80% of total hemicellulose in the biomass was separated out into the effluent primarily in the form of xylose oligomers. Decomposition of sugar components was insignificant. The remaining solids had a composition of 80–93% glucan, 5–10% xylan, and 1–6% lignin. Selected solid samples, obtained under near-optimum conditions, exhibited a chemical composition close to that of commercial α-cellulose The enzymatic digestibilities of these solid samples were substantially higher than that of α-cellulose.