The primary obstacle to producing renewable fuels from lignocellulosic biomass is a plant's recalcitrance to releasing sugars bound in the cell wall. From a sample set of wood cores representing 1,100 individual undomesticated Populus trichocarpa trees, 47 extreme phenotypes were selected across measured lignin content and ratio of syringyl and guaiacyl units (S/G ratio). This subset was tested for total sugar release through enzymatic hydrolysis alone as well as through combined hot-water pretreatment and enzymatic hydrolysis using a high-throughput screening method. The total amount of glucan and xylan released varied widely among samples, with total sugar yields of up to 92% of the theoretical maximum. A strong negative correlation between sugar release and lignin content was only found for pretreated samples with an S/G ratio < 2.0. For higher S/G ratios, sugar release was generally higher, and the negative influence of lignin was less pronounced. When examined separately, only glucose release was correlated with lignin content and S/G ratio in this manner, whereas xylose release depended on the S/G ratio alone. For enzymatic hydrolysis without pretreatment, sugar release increased significantly with decreasing lignin content below 20%, irrespective of the S/G ratio. Furthermore, certain samples featuring average lignin content and S/G ratios exhibited exceptional sugar release. These facts suggest that factors beyond lignin and S/G ratio influence recalcitrance to sugar release and point to a critical need for deeper understanding of cell-wall structure before plants can be rationally engineered for reduced recalcitrance and efficient biofuels production.

Building niches for multiple microbes: Microbial conversion of heterogeneous feedstocks such as lignocellulose into one or more desired products requires assembly of pathways to both break down the inputs and produce the outputs. Shahab et al. assembled a collection of microbes that occupy different spatial niches within a bioreactor and have different metabolic capabilities. In the simplest version of their setup, an aerobic fungus adeptly breaks down cellulose into short sugar chains, an oxygen-tolerant bacterium converts these to lactic acid, and an anaerobic bacterium uses the lactic acid to synthesize the short-chain fatty acid butyric acid. Additional microbes handle limitations in the intermediate stream or divert lactic acid to longer-chain fatty acids, which are potentially more valuable. The integrated production and use of lactic acid for biosynthetic reactions could ideally serve as a platform for biosynthesis using robust, heterogeneous microbial consortia. Science , this issue p. eabb1214

Steam explosion pretreatment has been examined in many studies for enhancing the enzymatic digestibility of lignocellulosic biomass and is currently the most common pretreatment method in commercial biorefineries. The information available about the effect of the explosive decompression on the biochemical conversion is, however, very limited, and no studies prove that the latter is actually enhanced by the explosion. Hence, it is of great value to discern between the effect of the explosion on the one hand and the steaming on the other hand, to identify their particular influences on enzymatic digestibility. The effect of the explosive decompression in the steam explosion pretreatment of spruce wood chips on their enzymatic cellulose digestibility was studied systematically. The explosion had a high influence on digestibility, improving it by up to 90 % compared to a steam pretreatment without explosion. Two factors were identified to be essentially responsible for the effect of the explosion on enzymatic digestibility: pretreatment severity and pressure difference of the explosion. A higher pretreatment severity can soften up and weaken the lignocellulose structure more, so that the explosion can better break up the biomass and decrease its particle size, which enhances its digestibility. In particular, increasing the pressure difference of the explosion leads to more defibration, a smaller particle size and a better digestibility. Though differences were found in the micro- and nanostructure of exploded and non-exploded biomass, the only influence of the explosion on digestibility was found to be the macroscopic particle size reduction. Steam explosion treatments with a high severity and a high pressure difference of the explosion lead to a comparatively high cellulose digestibility of the—typically very recalcitrant—softwood biomass. This is the first study to show that explosion can enhance the enzymatic digestibility of lignocellulosic biomass. If the enhancing effect of the explosion is thoroughly exploited, even very recalcitrant biomass like softwood can be made enzymatically digestible.

Lignocellulosic biomass – such as wood, agricultural residues or dedicated energy crops – is a promising renewable feedstock for production of fuels and chemicals that is available at large scale at low cost without direct competition for food usage. Its biochemical conversion in a sugar platform biorefinery includes three main unit operations that are illustrated in this review: the physico-chemical pretreatment of the biomass, the enzymatic hydrolysis of the carbohydrates to a fermentable sugar stream by cellulases and finally the fermentation of the sugars by suitable microorganisms to the target molecules. Special emphasis in this review is put on the technology, commercial status and future prospects of the production of second-generation fuel ethanol, as this process has received most research and development efforts so far. Despite significant advances, high enzyme costs are still a hurdle for large scale competitive lignocellulosic ethanol production. This could be overcome by a strategy termed 'consolidated bioprocessing' (CBP), where enzyme production, enzymatic hydrolysis and fermentation is integrated in one step – either by utilizing one genetically engineered superior microorganism or by creating an artificial co-culture. Insight is provided on both CBP strategies for the production of ethanol as well as of advanced fuels and commodity chemicals.

An autohydrolysis pretreatment that suppresses lignin repolymerisation helps overcoming the recalcitrance of softwood for enzymatic hydrolysis of its cellulose.

Lignocellulosic biomass is uniquely suited as a sustainable feedstock for the biotechnological production of alternative fuels and chemicals. However, due to the biomass recalcitrance, the enzymatic conversion process is complex and needs to be simplified. To this end, we developed a process, which allows the consolidated bioprocessing of lignocellulose to ethanol in a single multi-species biofilm membrane reactor featuring both aerobic and anaerobic conditions necessary for the simultaneous fungal cellulolytic enzyme production and alcoholic yeast fermentation of the hydrolysis-derived sugars. The general feasibility of the concept was successfully demonstrated by producing ethanol with a 67% yield from undetoxified whole slurry dilute acid pretreated wheat straw by the combined action of Trichoderma reesei, Saccharomyces cerevisiae and Scheffersomyces stipitis. The results achieved underscore the potential of the process as a versatile cheap sugar platform for the production of fuels and chemicals based on lignocellulosic biomass by specifically compiled consortia of industrially proven robust microorganisms.