Deconstruction Of Lignocellulosic Biomass Research Articles

Systematic bioprospection for cellulolytic actinomycetes in the Chihuahuan Desert: isolation and enzymatic profiling.

The quest for microbial cellulases has intensified as a response to global challenges in biofuel production. The efficient deconstruction of lignocellulosic biomass holds promise for generating valuable products in various industries such as food, textile, and detergents. This article presents a systematic bioprospection aimed at isolating actinomycetes with exceptional cellulose deconstruction capabilities. Our methodology explored the biodiverse oligotrophic region of Cuatro Cienegas, Coahuila, within the Chihuahuan Desert. Among the evaluated actinomycetes collection, 78% exhibited cellulolytic activity. Through a meticulous screening process based on enzymatic index evaluation, we identified a highly cellulolytic Streptomyces strain for further investigation. Submerged fermentation of this strain revealed an endoglucanase enzymatic activity of 149 U/mg. Genomic analysis of strain Streptomyces sp. STCH565-A revealed unique configurations of carbohydrate-active enzyme (CAZyme) genes, underscoring its potential for lignocellulosic bioconversion applications. These findings not only highlight the significance of the Chihuahuan Desert as a rich source of cellulolytic microorganisms but also offer insights into the systematic exploration and selection of high-performing cellulolytic microorganisms for application in diverse environmental contexts. In conclusion, our bioprospecting study lays a foundation for harnessing the cellulolytic potential of actinomycetes from the Chihuahuan Desert, with implications for advancing cellulose deconstruction processes in various industries. The findings can serve as a blueprint for future bioprospecting efforts in different regions, facilitating the targeted discovery of microorganisms with exceptional cellulosic deconstruction capabilities.

Exploring the cellulolytic and hemicellulolytic activities of manganese peroxidase for lignocellulose deconstruction

BackgroundA cost-effective pretreatment and saccharification process is a necessary prerequisite for utilizing lignocellulosic biomass (LCB) in biofuel and biomaterials production. Utilizing a multifunctional enzyme with both pretreatment and saccharification functions in a single step for simultaneous biological pretreatment and saccharification process (SPS) will be a green method of low cost and high efficiency. Manganese peroxidase (MnP, EC 1.11.1.13), a well-known lignin-degrading peroxidase, is generally preferred for the biological pretreatment of biomass. However, exploring the role and performance of MnP in LCB conversion will promote the application of MnP for lignocellulose-based biorefineries.ResultsIn this study, we explored the ability of an MnP from Moniliophthora roreri, MrMnP, in LCB degradation. With Mn2+ and H2O2, MrMnP decomposed 5.0 g/L carboxymethyl cellulose to 0.14 mM of reducing sugar with a conversion yield of 5.0 mg/g, including 40 μM cellobiose, 70 μM cellotriose, 20 μM cellotetraose, and 10 μM cellohexaose, and degraded 1.0 g/L mannohexaose to 0.33 μM mannose, 4.08 μM mannotriose, and 4.35 μM mannopentaose. Meanwhile, MrMnP decomposed 5.0 g/L lichenan to 0.85 mM of reducing sugar with a conversion yield of 30.6 mg/g, including 10 μM cellotriose, 20 μM cellotetraose, and 80 μM cellohexose independently of Mn2+ and H2O2. Moreover, the versatility of MrMnP in LCB deconstruction was further verified by decomposing locust bean gum and wheat bran into reducing sugars with a conversion yield of 54.4 mg/g and 29.5 mg/g, respectively, including oligosaccharides such as di- and tri-saccharides. The catalytic mechanism underlying MrMnP degraded lignocellulose was proposed as that with H2O2, MrMnP oxidizes Mn2+ to Mn3+. Subsequently, it forms a complex with malonate, facilitating the degradation of CMC and mannohexaose into reducing sugars. Without H2O2, MrMnP directly oxidizes malonate to hydroperoxyl acetic acid radical to form compound I, which then attacks the glucosidic bond of lichenan.ConclusionThis study identified a new function of MrMnP in the hydrolysis of cellulose and hemicellulose, suggesting that MrMnP exhibits its versatility in the pretreatment and saccharification of LCB. The results will lead to an in-depth understanding of biocatalytic saccharification and contribute to forming new enzymatic systems for using lignocellulose resources to produce sustainable and economically viable products and the long-term development of biorefinery, thereby increasing the productivity of LCB as a green resource.

Laccase-Based Self-Amplifying Catalytic System Enables Efficient Antibiotic Degradation for Sustainable Environmental Remediation.

Antibiotic contamination poses potential risks to ecosystems and human health. Laccase (LAC) has emerged as a promising biocatalyst for the oxidation of environmentally toxic contaminants with high catalytic efficiency; however, its large-scale application is hindered by enzyme costs and dependency on redox mediators. Herein, a novel self-amplifying catalytic system (SACS) for antibiotic remediation that does not require external mediators is developed. In SACS, a natural mediator-regenerating koji with high-activity LAC, derived from lignocellulosic waste, initiates the chlortetracycline (CTC) degradation. Subsequently, an intermediate product, CTC327, identified as an active mediator for LAC via molecular docking, is formed and then starts a renewable reaction cycle, including CTC327-LAC interaction, stimulated CTC bioconversion, and self-amplifying CTC327 release, thus enabling highly efficient antibiotic bioremediation. In addition, SACS exhibits excellent performance in producing lignocellulose-degrading enzymes, highlighting its potential for lignocellulosic biomass deconstruction. To demonstrate its effectiveness and accessibility in the natural environment, SACS is used to catalyze in situ soil bioremediation and straw degradation. The resulting CTC degradation rate is 93.43%, with a straw mass loss of up to 58.35% in a coupled process. This mediator regeneration and waste-to-resource conversion in SACS provides a promising route for environmental remediation and sustainable agricultural practices.

Contribution of calcium ligands in substrate binding and product release in the Acetovibrio thermocellus glycoside hydrolase family 9 cellulase CelR

Enzymatic deconstruction of lignocellulosic biomass is crucial to establishment of the renewable biofuel and bioproduct economy. Better understanding of these enzymes, including their catalytic and binding domains, and other features offer potential avenues for improvement. Glycoside hydrolase family 9 (GH9) enzymes are attractive targets because they have members that exhibit exo- and endo-cellulolytic activity, processivity of reaction, and thermostability. This study examines a GH9 from Acetovibrio thermocellus ATCC 27405, AtCelR containing a catalytic domain and a carbohydrate binding module (CBM3c). Crystal structures of the enzyme without substrate, bound to cellohexaose (substrate) or cellobiose (product), show the positioning of ligands to calcium and adjacent residues in the catalytic domain that may contribute to substrate binding and facilitate product release. We also investigated the properties of the enzyme engineered to contain an additional carbohydrate binding module (CBM3a). Relative to the catalytic domain alone, CBM3a gave improved binding for Avicel (a crystalline form of cellulose), and catalytic efficiency (kcat/KM) was improved 40×with both CBM3c and CBM3a present. However, because of the molecular weight added by CBM3a, the specific activity of the engineered enzyme was not increased relative to the native construct consisting of only the catalytic and CBM3c domains. This work provides new insight into a potential role of the conserved calcium in the catalytic domain and identifies contributions and limitations of domain engineering for AtCelR and perhaps other GH9 enzymes.

Pretreatment of poplar with eco-friendly levulinic acid to achieve efficient utilization of biomass

Organic acid pretreatment is an effective method for green separation of lignocellulosic biomass. However, repolymerization of lignin seriously affects the dissolution of hemicellulose and the conversion of cellulose during organic acid pretreatment. Therefore, a new organic acid pretreatment, levulinic acid (Lev) pretreatment, was studied for the deconstruction of lignocellulosic biomass without adding additional additives. The preferred separation of hemicellulose was realized at Lev concentration 7.0%, temperature 170 °C, and time 100 min. The separation of hemicellulose increased from 58.38% to 82.05% compared with acetic acid pretreatment. It was found that the repolymerization of lignin was effectively inhibited in the efficient separation of hemicellulose. This was attributed to the fact that γ-valerolactone (GVL) is a good green scavenger of lignin fragments. The lignin fragments in the hydrolysate were effectively dissolved. The results provided theoretical support for creating green and efficient organic acid pretreatment and effectively inhibiting lignin repolymerization.

Efficient co-production of fermentable sugars and biobutanol from corn stover based on a novel butyric acid pretreatment strategy

Efficient deconstruction of lignocellulosic biomass by pretreatment is important for the production of fermentable sugars and biofuels. This study used butyric acid as a novel catalyst for corn stover pretreatment. The effects of butyric acid pretreatments on chemical composition and enzymatic digestibility were systematically investigated. The results indicate that a high linear correlation between xylan removal and fermentable sugars yield (r2 = 0.91) was found. Pretreatment of corn stover with 50 g/L butyric acid at 180 °C for 0.5 h removed 83.17% of xylan and recovered 86.99% of glucan, and fermentable sugars yield of pretreated corn stover reached 97.16%. The physiochemical characterization results show that crystalline index (CrI) of corn stover was increased from 30.13% to 45.04%−48.38% after pretreatments, and the chemical structure and morphological features also exhibited positive changes for enhancing enzymatic hydrolysis. Meanwhile, the liquid fraction of pretreated slurry can be used for recycled pretreatment with 73.81% of fermentable sugars yield after four cycles. Additionally, the residual butyric acid was re-utilized as the co-substrate for Clostridium acetobutylicum ATCC 824 to produce biobutanol via acetone-butanol-ethanol (ABE) fermentation. The final production of butanol and solvents reached 8.38 g/L and 12.63 g/L under an optimized intermittent feeding strategy with a higher butanol/acetone ratio of 2.40 g/g. Overall, this work presents a novel pretreatment strategy to co-produce fermentable sugars and advanced biofuel from abundant lignocellulosic biomass.

Sustainable Strategies for the Conversion of Lignocellulosic Materials into Biohydrogen: Challenges and Solutions toward Carbon Neutrality

The present review mainly discusses advanced pretreatment techniques for converting lignocellulosic biomass into hydrogen. The focus of this review is also to acquire knowledge concerning lignocellulosic biomass pretreatment processes and their impact on the efficiency of biohydrogen fermentation. The deconstruction of lignocellulosic biomass is presented using various pretreatment techniques albeit with several advantages and disadvantages, particularly about the interference due to the generated inhibitory compounds is toxic to microbes used for fermentation. The use of an appropriate pretreatment process can make the recalcitrant lignocellulosic biomass substrates amenable for further microbial fermentation to produce hydrogen. Although till date there is no ideal pretreatment step available to develop a cost-effective process for conversion of lignocellulosic materials into fermentable sugars, nanotechnology seem to be a more sustainable approach as compared to the traditional processes.

PACER: a novel 3D plant cell wall model for the analysis of non-catalytic and enzymatic responses

BackgroundSubstrate accessibility remains a key limitation to the efficient enzymatic deconstruction of lignocellulosic biomass. Limited substrate accessibility is often addressed by increasing enzyme loading, which increases process and product costs. Alternatively, considerable efforts are underway world-wide to identify amorphogenesis-inducing proteins and protein domains that increase the accessibility of carbohydrate-active enzymes to targeted lignocellulose components.ResultsWe established a three-dimensional assay, PACER (plant cell wall model for the analysis of non-catalytic and enzymatic responses), that enables analysis of enzyme migration through defined lignocellulose composites. A cellulose/azo-xylan composite was made to demonstrate the PACER concept and then used to test the migration and activity of multiple xylanolytic enzymes. In addition to non-catalytic domains of xylanases, the potential of loosenin-like proteins to boost xylanase migration through cellulose/azo-xylan composites was observed.ConclusionsThe PACER assay is inexpensive and parallelizable, suitable for screening proteins for ability to increase enzyme accessibility to lignocellulose substrates. Using the PACER assay, we visualized the impact of xylan-binding modules and loosenin-like proteins on xylanase mobility and access to targeted substrates. Given the flexibility to use different composite materials, the PACER assay presents a versatile platform to study impacts of lignocellulose components on enzyme access to targeted substrates.

Microbial conversion of lignin rich biomass hydrolysates to medium chain length polyhydroxyalkanoates (mcl-PHA) using Pseudomonas putida KT2440

As world moves toward increasing number of products being produced from renewable lignocellulosic agricultural and forest residues, the major classes of products that will shift to greener routes on priority are energy, fuels, and materials in that order. In materials segment, polyhydroxyalkanoates are an emerging class of biopolyesters with several potential industrial uses. The present work investigates medium chain length polyhydroxyalkanoates (mcl-PHA) producing capabilities of Pseudomonas putida KT2440 from a mixture of compounds produced from lignocellulosic biomass deconstruction. The hydrolysates obtained from nitric acid pretreatment of lignin rich cotton stalk (CS) and palm empty fruit bunch (EFB) were used as substrates for production of mcl-PHA. Presence of 3-hydroxydecanoate and 3-hydroxyocytanoate observed on GC-MS confirmed PHA accumulation in the cells. PHA accumulation was estimated between 20% and 35% of cell dry weight when grown on both model substrates as well as biomass hydrolysates. PHA titers obtained on hydrolysates of CS and EFB were 0.24 g/L and 0.21 g/L, respectively.

Pathways of lignocellulosic biomass deconstruction for biofuel and value-added products production

As the world attempts to transition from fossil fuels, lignocellulosic biomass (LCB) serves as a promising alternative due to its high abundance. Hydrolysing LCB can generate various bioproducts, such as biofuels and value-added chemicals. However, the presence of lignin inhibits the solubilization of LCBs, presenting a major techno-economic challenge in the biorefinery concept. Therefore, this paper addresses the gaps left by most of the recent review works that fail to comprehensively review different pretreatment methods and the full scope of applications of LCBs, and do not incorporate techno-economic considerations of the technologies, the latter being the greatest bottleneck in the commercialization of the processes. The literature review revealed that while many of the physical and chemical pretreatment methods exhibit great effectiveness, they have a huge dependence on energy, chemicals, water, and/or specialized equipment, and produce harmful waste and inhibitory compounds. The pretreatment of lignocellulosic biomass can account for 40% of total production costs. Biological pretreatment can address these challenges but is limited by long incubation times. For instance, the bacterial pretreatment can noticeably reduce sawdust cellulose, hemicelluloses, and lignin contents by 35.8%, 37.1%, and 46.2%, respectively. Recently, integrated/coupling (hybrid) methods, such as chemical-assisted liquid hot water/steam and microwave or ultrasound-assisted alkaline pretreatment, have been gaining popularity due to their potential to improve chemical yield, but at the expense of the high cost of operation. To make pretreatment processes more techno-economically feasible, there is a need for process integration and the standardization and optimization of process parameters.

Evaluation of Exogenous Laccase Enzyme Treated Finger Millet Straw on Body Weight Gain, DM Intake and Nutrient Digestibility in Heifers

Background: Fungal laccases have widespread application in a number of biotechnological processes including the biodegradation of lignin. Their low yield in the native state limits their practical use in the deconstruction of lignocellulosic biomass for feeding ruminants. Enzymes in bulk quantities are required to treat biomass which has got greater product consistency and less lot to lot variations. The present study was an attempt to evaluate the effect of feeding finger millet straw treated with exogenous laccases in heifers.Methods: The effect of feeding exogenous laccases obtained from immobilized Schizophyllum commune (MTCC 11893) on body weight gain, dry matter intake and nutrient digestibility in heifers was evaluated in three groups of heifers (4-each) with body weights ranging between 275 ± 47.12 to 276.75 ± 64.48kg. The control group received ad lib. finger millet straw treated with only production media (GI). Test group 1 (GII) received ad lib. straw treated with laccase rich media in a 3:5 (w/v) ratio, while test group 2 (GIII) received ad lib. straw treated in a 4:5 (w/v) ratio. Result: After 14- days of feeding, gain in body weights for GI, GII and GIII were 277.9 ± 68.47; 277.50 ± 46.43 and 278.85 ± 37.22 respectively with an overall increase of 1.15, 4.25 and 3.60 kgs. No significant variation (P greater than 0.05) was observed with regard to DMD (%) between the groups which was 46 ± 5.8 for the control animals and 41±13.0 and 41± 3.4 in G2 and G3 groups. Though significant variations were observed digestibility studies proved inconclusive. Though preliminary results indicate that applying lignin degrading enzymes as feed supplements could enhance digestibility of crop residues in ruminants.

Iron incorporation both intra- and extra-cellularly improves the yield and saccharification of switchgrass (Panicum virgatum L.) biomass

BackgroundPretreatments are commonly used to facilitate the deconstruction of lignocellulosic biomass to its component sugars and aromatics. Previously, we showed that iron ions can be used as co-catalysts to reduce the severity of dilute acid pretreatment of biomass. Transgenic iron-accumulating Arabidopsis and rice plants exhibited higher iron content in grains, increased biomass yield, and importantly, enhanced sugar release from the biomass.ResultsIn this study, we used intracellular ferritin (FerIN) alone and in combination with an improved version of cell wall-bound carbohydrate-binding module fused iron-binding peptide (IBPex) specifically targeting switchgrass, a bioenergy crop species. The FerIN switchgrass improved by 15% in height and 65% in yield, whereas the FerIN/IBPex transgenics showed enhancement up to 30% in height and 115% in yield. The FerIN and FerIN/IBPex switchgrass had 27% and 51% higher in planta iron accumulation than the empty vector (EV) control, respectively, under normal growth conditions. Improved pretreatability was observed in FerIN switchgrass (~ 14% more glucose release than the EV), and the FerIN/IBPex plants showed further enhancement in glucose release up to 24%.ConclusionsWe conclude that this iron-accumulating strategy can be transferred from model plants and applied to bioenergy crops, such as switchgrass. The intra- and extra-cellular iron incorporation approach improves biomass pretreatability and digestibility, providing upgraded feedstocks for the production of biofuels and bioproducts.

Molecular Insight into Glucose-Induced Conformational Change to Investigate Uncompetitive Inhibition of GH1 β-Glucosidase

β-glucosidase-catalyzed cellobiose conversion to glucose is the principal rate-limiting step in the deconstruction of lignocellulosic biomass to biofuel production as most β-glucosidases are feedba...

Improvement of potassium permanganate pretreatment by enzymatic saccharification of rice straw for production of biofuels

Commonly, the agricultural waste, i.e. lignocellulosic biomass is disposed through combustion causing air pollution with production of PM2.5 and PM10 particles. However, it has been found that these biomasses can be used as source for the production of biofuels and other valuable biochemicals. Though deconstruction of lignocellulosic biomass is challenging due to its complex structure. In this study, rice straw (RS) was pretreated using potassium permanganate (KMnO4) to enhance the enzymatic saccharification efficiency. The study was carried out by varying the operational factors in pretreatment, including temperature (30-90°C), time (30-360 min) and concentration of KMnO4 (0.5-3.0, % w/v), respectively, based on Box-Behnken design (BBD). Through multi-regression analysis of the experimental data obtained after pretreatment, the optimum conditions were determined. The optimum conditions for temperature, time and potassium permanganate concentration were 48.09°C, 360 min, and 1.36% w/v, respectively. The saccharifications of pretreatment and untreated rice straw were carried out using Cellic Ctec2. The reducing sugar was determined by using DNS method and the yields of the untreated and pretreated RS were 32.38 and 49.011 mg/mL, respectively. The results showed that the sugar for pretreated RS were 1.51 fold times higher compared to untreated RS. Therefore, this work illustrates the pretreatment efficiency for KMnO4 to enhance the reducing sugar yield during saccharification, which can be used for biofuel and value-added product productions.

Availability of the Molecular Switch XylR Controls Phenotypic Heterogeneity and Lag Duration during Escherichia coli Adaptation from Glucose to Xylose.

ABSTRACTThe glucose-xylose metabolic transition is of growing interest as a model to explore cellular adaption since these molecules are the main substrates resulting from the deconstruction of lignocellulosic biomass. Here, we investigated the role of the XylR transcription factor in the length of the lag phases when the bacterium Escherichia coli needs to adapt from glucose- to xylose-based growth. First, a variety of lag times were observed when different strains of E. coli were switched from glucose to xylose. These lag times were shown to be controlled by XylR availability in the cells with no further effect on the growth rate on xylose. XylR titration provoked long lag times demonstrated to result from phenotypic heterogeneity during the switch from glucose to xylose, with a subpopulation unable to resume exponential growth, whereas the other subpopulation grew exponentially on xylose. A stochastic model was then constructed based on the assumption that XylR availability influences the probability of individual cells to switch to xylose growth. The model was used to understand how XylR behaves as a molecular switch determining the bistability set-up. This work shows that the length of lag phases in E. coli is controllable and reinforces the role of stochastic mechanism in cellular adaptation, paving the way for new strategies for the better use of sustainable carbon sources in bioeconomy.

Three-Dimensional Imaging of Plant Cell Wall Deconstruction Using Fluorescence Confocal Microscopy

Lignocellulosic biomass (LB) is recalcitrant to enzymatic hydrolysis due to its compact and complex cell wall structure. To identify the parameters behind LB recalcitrance, experimental data over hydrolysis time must be collected. Here, we describe a novel method to collect time-lapse images during cell wall deconstruction by enzymatic hydrolysis. The protocol includes instructions for sample preparation, layout of a custom designed incubation chamber and instructions for confocal time lapse acquisition. The protocol sets out a detailed plan where cross-sections of untreated and pretreated poplar samples are mounted in a sealed frame containing a buffer and an enzymatic cocktail. The sealed frame is then placed into an incubator to maintain the sample at a constant temperature of 50 °C, which is optimal for enzymatic reaction while avoiding enzymatic cocktail evaporation. Using lignin natural autofluorescence, confocal z-stacks of untreated and pretreated samples were acquired at regular time intervals during enzymatic hydrolysis for 24 h. Acquisition parameters were optimized to compromise between image resolution and reduced photo-bleaching. The acquired image might then be processed by further development of algorithms to extract precise quantitative information on cell wall deconstruction. This protocol is an important first step towards elucidating the underlying parameters of LB recalcitrance by allowing the acquisition of high-quality images of LB hydrolysis for extracting quantitative data on LB deconstruction.

Endogenous carbohydrate esterases of Clostridium thermocellum are identified and disrupted for enhanced isobutyl acetate production from cellulose.

Medium-chain esters are versatile chemicals with broad applications as flavors, fragrances, solvents, and potential drop-in biofuels. Currently, these esters are largely produced by the conventional chemical process that uses harsh operating conditions and requires high energy input. Alternatively, the microbial conversion route has recently emerged as a promising platform for sustainable and renewable ester production. The ester biosynthesis pathways can utilize either lipases or alcohol acyltransferase (AAT), but the AAT-dependent pathway is more thermodynamically favorable in an aqueous fermentation environment. Even though a cellulolytic thermophile Clostridium thermocellum harboring an AAT-dependent pathway has recently been engineered for direct conversion of lignocellulosic biomass into esters, the production is not efficient. One potential bottleneck is the ester degradation caused by the endogenous carbohydrate esterases (CEs) whose functional roles are poorly understood. The challenge is to identify and disrupt CEs that can alleviate ester degradation while not negatively affecting the efficient and robust capability of C. thermocellum for lignocellulosic biomass deconstruction. In this study, by using bioinformatics, comparative genomics, and enzymatic analysis to screen a library of CEs, we identified and disrupted the two most critical CEs, Clo1313_0613 and Clo1313_0693, that significantly contribute to isobutyl acetate degradation in C. thermocellum. We demonstrated that an engineered esterase-deficient C. thermocellum strain not only reduced ester hydrolysis but also improved isobutyl acetate production while maintaining effective cellulose assimilation.

Eucalyptus red grandis pretreatment with protic ionic liquids: effect of severity and influence of sub/super-critical CO2 atmosphere on pretreatment performance.

Deconstruction of lignocellulosic biomass with low-cost ionic liquids (ILs) has proven to be a promising technology that could be implemented in a biorefinery to obtain renewable materials, fuels and chemicals. This study investigates the pretreatment efficacy of the ionoSolv pretreatment of Eucalyptus red grandis using the low-cost ionic liquid triethylammonium hydrogen sulfate ([N2220][HSO4]) in the presence of 20 wt% water at 10% solids loading. The temperatures investigated were 120 °C and 150 °C. Also, the influence of performing the pretreatment under sub-critical and supercritical CO2 was investigated. The IL used is very effective in deconstructing eucalyptus, producing cellulose-rich pulps resulting in enzymatic saccharification yields of 86% for some pretreatment conditions. It has been found that under a CO2 atmosphere, the ionoSolv process is pressure independent. The good performance of this IL in the pretreatment of eucalyptus is promising for the development of a large-scale ionoSolv pretreatment processes.

Mining the biomass deconstructing capabilities of rice yellow stem borer symbionts

BackgroundEfficient deconstruction of lignocellulosic biomass into simple sugars in an economically viable manner is a prerequisite for its global acceptance as a feedstock in bioethanol production. This is achieved in nature by suites of enzymes with the capability of efficiently depolymerizing all the components of lignocellulose. Here, we provide detailed insight into the repertoire of enzymes produced by microorganisms enriched from the gut of the crop pathogen rice yellow stem borer (Scirpophaga incertulas).ResultsA microbial community was enriched from the gut of the rice yellow stem borer for enhanced rice straw degradation by sub-culturing every 10 days, for 1 year, in minimal medium with rice straw as the main carbon source. The enriched culture demonstrated high cellulolytic and xylanolytic activity in the culture supernatant. Metatranscriptomic and metaexoproteomic analysis revealed a large array of enzymes potentially involved in rice straw deconstruction. The consortium was found to encode genes ascribed to all five classes of carbohydrate-active enzymes (GHs, GTs, CEs, PLs, and AAs), including carbohydrate-binding modules (CBMs), categorized in the carbohydrate-active enzymes (CAZy) database. The GHs were the most abundant class of CAZymes. Predicted enzymes from these CAZy classes have the potential to digest each cell-wall components of rice straw, i.e., cellulose, hemicellulose, pectin, callose, and lignin. Several identified CAZy proteins appeared novel, having an unknown or hypothetical catalytic counterpart with a known class of CBM. To validate the findings, one of the identified enzymes that belong to the GH10 family was functionally characterized. The enzyme expressed in E. coli efficiently hydrolyzed beechwood xylan, and pretreated and untreated rice straw.ConclusionsThis is the first report describing the enrichment of lignocellulose degrading bacteria from the gut of the rice yellow stem borer to deconstruct rice straw, identifying a plethora of enzymes secreted by the microbial community when growing on rice straw as a carbon source. These enzymes could be important candidates for biorefineries to overcome the current bottlenecks in biomass processing.

Approaches for More Efficient Biological Conversion of Lignocellulosic Feedstocks to Biofuels and Bioproducts

The future bioeconomy promises drop-in or performance-advantaged biofuels and bioproducts derived from lignocellulosic biomass, substantial greenhouse gas emissions reductions in sectors with few or no alternatives, and increased domestic energy production in countries with sufficient biomass resources. Despite the slower than anticipated pace of commercializing next-generation biofuels, the research community continues to make dramatic improvements at every stage of production, from feedstock cultivation through conversion to final products. However, the interdisciplinary nature of bioenergy research, and the need for cross-coordination among biologists, chemists, agronomists, and engineers, make coordinating and optimizing these strategies challenging. This Perspective surveys recent advancements in bioenergy crop engineering, lignocellulosic biomass deconstruction and fractionation, catabolism of biomass-derived sugars and aromatics, and biological conversion to fuels and products. We organize major research approaches into broad categories and comment on which strategies offer synergies or trade-offs in the context of four approaches to improving the economics and carbon-efficiency of advanced biofuels and bioproducts: (1) maximize sugar conversion to a single product, (2) utilize diverse carbon sources for producing a single product, (3) convert lignin to high-value products, and (4) fractionate the hydrolysate to derive maximum value from each component.