Producing concentrated sugars and minimizing water usage are key elements in the economics and environmental sustainability of advanced biofuels. Conventional pretreatment processes that require a water-wash step can result in losses of fermentable sugars and generate large volumes of wastewater or solid waste. To address these problems, we have developed high gravity biomass processing with a one-pot conversion technology that includes ionic liquid pretreatment, enzymatic saccharification, and yeast fermentation for the production of concentrated fermentable sugars and high-titer cellulosic ethanol. The use of dilute bio-derived ionic liquids (a.k.a. bionic liquids) enables one-pot, high-gravity bioethanol production due to their low toxicity to the hydrolytic enzyme mixtures and microbes used. We increased biomass digestibility at >30 wt% by understanding the relationship between ionic liquid and biomass loading, yielding 41.1 g L -1 of ethanol (equivalent to an overall yield of 74.8% on a glucose basis)using an integrated one-pot fed-batch system. Our technoeconomic analysis indicates that the optimized one-pot configuration provides significant economic and environmental benefits for cellulosic biorefineries by reducing the amount of ionic liquid required by ~90% and pretreatment-related water inputs and wastewater generation by ~85%. In turn, these improvements can reduce net electricity use, greenhouse gas-intensive chemical inputs for wastewater treatment, and waste generation. The result is an overall 40% reduction in the cost of cellulosic ethanol produced and a reduction in local burdens on water resources and waste management infrastructure.

We present an inexpensive and biocompatible protic ionic liquid that enables one-pot integrated cellulosic ethanol production without any pH adjustments and without water-wash or solid–liquid separations.

In future biorefineries, the development of inexpensive and renewable reagents for biomass pretreatment is highly desirable.

BackgroundEconomical conversion of lignocellulosic biomass into biofuels and bioproducts is central to the establishment of a robust bioeconomy. This requires a conversion host that is able to both efficiently assimilate the major lignocellulose-derived carbon sources and divert their metabolites toward specific bioproducts.ResultsIn this study, the carotenogenic yeast Rhodosporidium toruloides was examined for its ability to convert lignocellulose into two non-native sesquiterpenes with biofuel (bisabolene) and pharmaceutical (amorphadiene) applications. We found that R. toruloides can efficiently convert a mixture of glucose and xylose from hydrolyzed lignocellulose into these bioproducts, and unlike many conventional production hosts, its growth and productivity were enhanced in lignocellulosic hydrolysates relative to purified substrates. This organism was demonstrated to have superior growth in corn stover hydrolysates prepared by two different pretreatment methods, one using a novel biocompatible ionic liquid (IL) choline α-ketoglutarate, which produced 261 mg/L of bisabolene at bench scale, and the other using an alkaline pretreatment, which produced 680 mg/L of bisabolene in a high-gravity fed-batch bioreactor. Interestingly, R. toruloides was also observed to assimilate p-coumaric acid liberated from acylated grass lignin in the IL hydrolysate, a finding we verified with purified substrates. R. toruloides was also able to consume several additional compounds with aromatic motifs similar to lignin monomers, suggesting that this organism may have the metabolic potential to convert depolymerized lignin streams alongside lignocellulosic sugars.ConclusionsThis study highlights the natural compatibility of R. toruloides with bioprocess conditions relevant to lignocellulosic biorefineries and demonstrates its ability to produce non-native terpenes.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-017-0927-5) contains supplementary material, which is available to authorized users.

Naturally, many aerobic organisms degrade lignin-derived aromatics through conserved intermediates including protocatechuate and catechol. Employing this microbial approach offers a potential solution for valorizing lignin into valuable chemicals for a potential lignocellulosic biorefinery and enabling bioeconomy. In this study, two hybrid biochemical routes combining lignin chemical depolymerization, plant metabolic engineering, and synthetic pathway reconstruction were demonstrated for valorizing lignin into value-added products. In the biochemical route 1, alkali lignin was chemically depolymerized into vanillin and syringate as major products, which were further bio-converted into cis, cis-muconic acid (ccMA) and pyrogallol, respectively, using engineered Escherichia coli strains. In the second biochemical route, the shikimate pathway of Tobacco plant was engineered to accumulate protocatechuate (PCA) as a soluble intermediate compound. The PCA extracted from the engineered Tobacco was further converted into ccMA using the engineered E. coli strain. This study reports a direct process for converting lignin into ccMA and pyrogallol as value-added chemicals, and more importantly demonstrates benign methods for valorization of polymeric lignin that is inherently heterogeneous and recalcitrant. Our approach also validates the promising combination of plant engineering with microbial chassis development for the production of value added and speciality chemicals.

A catalytic amount of fluoride salt is all that is required to generate high molecular weight poly(phenylene ethynylene)s from silylacetylene-functionalized monomers and C6F6. The sole side product is gaseous fluorotrimethylsilane and the catalyst is removed simply by washing with water. The polymerization proceeds through a reactive intermediate, causing significant deviation from theory describing the relationship between comonomer stoichiometry and degree of polymerization for classical step-growth polymerizations.

Delignification as a function of ionic liquid (IL) pretreatment has potential in terms of recovering and converting the fractionated lignin streams to renewable products. Renewable biogenic ionic liquids, or bionic liquids (e.g., cholinium lysinate, ([Ch][Lys])), provide opportunities in terms of effective, economic, and sustainable lignocellulosic biomass pretreatment. We have evaluated [Ch][Lys] pretreatment in terms of sugar and lignin yields for three different feedstocks: switchgrass, eucalyptus, and pine. Four lignin streams isolated during [Ch][Lys] pretreatment and enzymatic hydrolysis were comprehensively analyzed, tracking their changes in physical−chemical structures. We observed changes in major lignin linkages and lignin aromatics units (phydroxyphenyl (H), guaiacyl (G), and syringil (S)) that occurred during pretreatment. A compositional analysis of the different process streams and a comprehensive mass balance in conjunction with multiple analytical techniques (nuclear magnetic resonance (NMR), mass spectroscopy, gel permeation chromatography (GPC)) is presented. Qualitative and quantitative analyses indicates that there are significantly more lignin−carbohydrate interactions for G-rich lignin in pine. The lignin removal and extent of lignin depolymerization for switchgrass and eucalyptus were higher than pine and follows the order of switchgrass > eucalyptus > pine. The insights gained from this work contribute to better understanding of physiochemical properties of lignin streams generated during [Ch][Lys] pretreatment, offering a starting point for lignin valorization strategies.

A detailed study of chemical changes in lignin structure during the ionic liquid (IL) pretreatment process is not only pivotal for understanding and overcoming biomass recalcitrance during IL pretreatment but is also necessary for designing new routes for lignin valorization. Chemical changes in lignin were systematically studied as a function of pretreatment temperature, time, and type of IL used. Kraft lignin was used as the lignin source, and common pretreatment conditions were employed using three different ILs of varying chemical structure in terms of acidic or basic character. The chemical changes in the lignin structure due to IL pretreatment processes were monitored using 1H–13C heteronuclear single quantum coherence (HSQC) nuclear magnetic resonance (NMR), 31P NMR, elemental analysis, gel permeation chromatography (GPC), and Fourier transform infrared (FT-IR), and the depolymerized products were analyzed using gas chromatography mass spectrometry (GC-MS). Although, with pretreatment in acidic IL, triethyl­ammonium hydrogensulfate ([TEA]­[HSO4]) results in the maximum decrease in the β-aryl ether bond, maximum dehydration and recondensation pathways were also evident, with the net process showing a minimum decrease in the molecular weight of regenerated lignin. However, 1-ethyl-3-methyl­imidazolium acetate ([C2C1Im]­[OAc]) pretreatment yields a smaller decrease in the β-aryl ether content along with minimum evidence of recondensation, resulting in the maximum decrease in the molecular weight. Cholinium lysinate ([Ch]­[Lys]) pretreatment shows an intermediate result, with moderate depolymerization, dehydration, and recondensation observed. The depolymerization products after IL pretreatment are found to be a function of the pretreatment temperature and the specific chemical nature of the IL used. At higher pretreatment temperature, [Ch]­[Lys] pretreatment yields guaiacol, [TEA]­[HSO4] yields guaiacylacetone, and [C2C1Im]­[OAc] yields both guaiacol and guaiacylacetone as major products. These results clearly indicate that the changes in lignin structure as well as the depolymerized product profile depend on the pretreatment conditions and the nature of the ILs. The insight gained on lignin structure changes and possible depolymerized products during IL pretreatment process would help future lignin valorization efforts in a potential IL-based lignocellulosic biorefinery.