Lignocellulosic Plant Biomass Research Articles

The Technology Used for Synthetic Polyploid Production of Miscanthus as Cellulosic Biofuel Feedstock

Background: The contemporary bioethanol production technologies are based on the utilization of plant lignocellulosic biomass. These technologies require conducting regular search, breeding, and creation of new energy crops. In particular, significant attention is paid to plants of the genus Miscanthus - perennial grasses that have a great potential as renewable energy sources. The main advantages of representatives of this species are considered to be high biomass yield, cold tolerance, a low requirement to soil conditions, long-term use of plantation, etc. M. × giganteus is the most promising species of the genus. The last is a sterile allotriploid originated through the hybridization of M. sinensis and M. sacchariflorus. Due to the problem with sexual reproduction, there is a lack of genotypes necessary for plant breeding programs to improve this species. Thus, polyploidization is an indispensable approach for obtaining new genotypes of M. × giganteus. Objective: The aim of this work is to review the attempts and methodologies employed to induce polyploidy in plants belonging to the genus Miscanthus. Methods: For this purpose, the concentrations and duration of treatment with different antimitotic agents in species, within this genus, have been considered. Methods for ploidy level determination and evaluation of biological and biochemical traits in the resulting polyploids have also been reported. Results: The application of antimitotic agents in vitro is the most effective and commonly used method of polyploidization in Miscanthus. The most effective antimitotic compounds appeared to be the well-known dinitroanilines, such as oryzalin and trifluralin, as well as new dinitroanilines with significantly lower phytotoxicity level. Conclusion: Polyploidization in Miscanthus has been investigated by various research groups worldwide. Currently, polyploid forms of M. sinensis, M. sacchariflorus and M. × giganteus have been obtained. The biological and biochemical traits of the obtained polyploids differ significantly from their original forms. However, the challenge of fertility restoration of M. × giganteus has not been resolved yet.

Towards a Miniaturized Culture Screening for Cellulolytic Fungi and Their Agricultural Lignocellulosic Degradation.

The substantial use of fungal enzymes to degrade lignocellulosic plant biomass has widely been attributed to the extensive requirement of powerful enzyme-producing fungal strains. In this study, a two-step screening procedure for finding cellulolytic fungi, involving a miniaturized culture method with shake-flask fermentation, was proposed and demonstrated. We isolated 297 fungal strains from several cellulose-containing samples found in two different locations in Thailand. By using this screening strategy, we then selected 9 fungal strains based on their potential for cellulase production. Through sequence-based identification of these fungal isolates, 4 species in 4 genera were identified: Aspergillus terreus (3 strains: AG466, AG438 and AG499), Penicillium oxalicum (4 strains: AG452, AG496, AG498 and AG559), Talaromyces siamensis (1 strain: AG548) and Trichoderma afroharzianum (1 strain: AG500). After examining their lignocellulose degradation capacity, our data showed that P. oxalicum AG452 exhibited the highest glucose yield after saccharification of pretreated sugarcane trash, cassava pulp and coffee silverskin. In addition, Ta. siamensis AG548 produced the highest glucose yield after hydrolysis of pretreated sugarcane bagasse. Our study demonstrated that the proposed two-step screening strategy can be further applied for discovering potential cellulolytic fungi isolated from various environmental samples. Meanwhile, the fungal strains isolated in this study will prove useful in the bioconversion of agricultural lignocellulosic residues into valuable biotechnological products.

Superior cellulolytic activity of Trichoderma guizhouense on raw wheat straw.

Lignocellulosic plant biomass is the world's most abundant carbon source and has consequently attracted attention as a renewable resource for production of biofuels and commodity chemicals that could replace fossil resources. Due to its recalcitrant nature, it must be pretreated by chemical, physical or biological means prior to hydrolysis, introducing additional costs. In this paper, we tested the hypothesis that fungi which thrive on lignocellulosic material (straw, bark or soil) would be efficient in degrading untreated lignocellulose. Wheat straw was used as a model. We developed a fast and simple screening method for cellulase producers and tested one hundred Trichoderma strains isolated from wheat straw. The most potent strain-UB483FTG2/ TUCIM 4455, was isolated from substrate used for mushroom cultivation and was identified as T. guizhouense. After optimization of growth medium, high cellulase activity was already achieved after 72h of fermentation on raw wheat straw, while the model cellulase overproducing strain T. reesei QM 9414 took 170h and reached only 45% of the cellulase activity secreted by T. guizhouense. Maximum production levels were 1.1 U/mL (measured with CMC as cellulase substrate) and 0.7 U/mL (β-glucosidase assay). The T. guizhouense cellulase cocktail hydrolyzed raw wheat straw within 35h. Our study shows that screening for fungi that successfully compete for special substrates in nature will lead to the isolation of strains with qualitatively and quantitatively superior enzymes needed for their digestion which could be used for industrial purposes.

Fine Comminution of Pine Bark: How Does Mechanical Loading Influence Particles Properties and Milling Efficiency?

The use of lignocellulosic plant biomass as an alternative to fossil feedstocks for chemistry, energy and materials often involves an intense dry comminution step, for which the energy consumed can vary significantly according to the process parameters, the particle size targeted, and the properties of the biomass. Here we studied the fine milling of maritime pine bark in an impact-mill configuration and in an attrition-mill configuration. The properties of the resulting powders (particle size distribution, particle shape, specific surface area, agglomeration level) obtained in each configuration were compared in relation to process energy consumption. Results evidenced that the agglomeration phenomena drive milling efficiency and limit the possibilities for reaching ultrafine particles. Interestingly, impact loading proved more effective at breaking down coarse particles but tended to generate high agglomeration levels, whereas attrition milling led to less agglomeration and thus to finer particles.

The White-Rot Basidiomycete Dichomitus squalens Shows Highly Specific Transcriptional Response to Lignocellulose-Related Aromatic Compounds.

Lignocellulosic plant biomass is an important feedstock for bio-based economy. In particular, it is an abundant renewable source of aromatic compounds, which are present as part of lignin, as side-groups of xylan and pectin, and in other forms, such as tannins. As filamentous fungi are the main organisms that modify and degrade lignocellulose, they have developed a versatile metabolism to convert the aromatic compounds that are toxic at relatively low concentrations to less toxic ones. During this process, fungi form metabolites some of which represent high-value platform chemicals or important chemical building blocks, such as benzoic, vanillic, and protocatechuic acid. Especially basidiomycete white-rot fungi with unique ability to degrade the recalcitrant lignin polymer are expected to perform highly efficient enzymatic conversions of aromatic compounds, thus having huge potential for biotechnological exploitation. However, the aromatic metabolism of basidiomycete fungi is poorly studied and knowledge on them is based on the combined results of studies in variety of species, leaving the overall picture in each organism unclear. Dichomitus squalens is an efficiently wood-degrading white-rot basidiomycete that produces a diverse set of extracellular enzymes targeted for lignocellulose degradation, including oxidative enzymes that act on lignin. Our recent study showed that several intra- and extracellular aromatic compounds were produced when D. squalens was cultivated on spruce wood, indicating also versatile aromatic metabolic abilities for this species. In order to provide the first molecular level systematic insight into the conversion of plant biomass derived aromatic compounds by basidiomycete fungi, we analyzed the transcriptomes of D. squalens when grown with 10 different lignocellulose-related aromatic monomers. Significant differences for example with respect to the expression of lignocellulose degradation related genes, but also putative genes encoding transporters and catabolic pathway genes were observed between the cultivations supplemented with the different aromatic compounds. The results demonstrate that the transcriptional response of D. squalens is highly dependent on the specific aromatic compounds present suggesting that instead of a common regulatory system, fine-tuned regulation is needed for aromatic metabolism.

Manganese Deficiency Is Required for High Itaconic Acid Production From D-Xylose in Aspergillus terreus.

Itaconic acid is used as a bio-based, renewable building block in the polymer industry. It is produced by submerged fermentations of the filamentous fungus Aspergillus terreus from molasses or starch, but research over the efficient utilization of non-food, lignocellulosic plant biomass is soaring. The objective of this study was to test whether the application of two key cultivation parameters for obtaining itaconic acid from D-glucose in high yields – Mn2+ ion deficiency and high concentration of the carbon source – would also occur on D-xylose, the principal monomer of lignocellulose. To this end, a carbon and energy balance for itaconic acid formation was established, which is 0.83 moles/mole D-xylose. The effect of Mn2+ ions on itaconic acid formation was indeed similar to that on D-glucose and maximal yields were obtained below 3 μg L–1 Mn2+ ions, which were, however, only 0.63 moles of itaconic acid per mole D-xylose. In contrast to the case on D-glucose, increasing D-xylose concentration over 50 g L–1 did not change the above yield. By-products such as xylitol and α-ketoglutarate were found, but in total they remained below 2% of the concentration of D-xylose. Mass balance of the fermentation with 110 g L–1 D-xylose revealed that >95% of the carbon from D-xylose was accounted as biomass, itaconic acid, and the carbon dioxide released in the last step of itaconic acid biosynthesis. Our data show that the efficiency of biomass formation is the critical parameter for itaconic acid yield from D-xylose under otherwise optimal conditions.

Modifications of microcrystalline cellulose (MCC), nanofibrillated cellulose (NFC), and nanocrystalline cellulose (NCC) for antimicrobial and wound healing applications

AbstractRecently, great attention has been paid to nano-composites of cellulose, due to their unique structure as a most abundant natural polymer with having exceptional properties such as renewable, biodegradable and high specific tensile strength, aspect ratio, and Young’s modulus. Prominent cellulose is naturally present in plant lignocellulosic biomass as a biocomposite made of cellulose, hemi-celluloses, lignin, etc. In addition, it can be extracted from other natural sources including bacteria, algae, and sea animals. Microcrystalline cellulose (MCC), nanocrystalline cellulose (NCC), and nanofibrillated cellulose (NFC) is an emerging renewable nanomaterial that has various applications, such as food, paper production, industrial and pharmaceutical biomaterials. The surface modification on NCC can improves its disperse ability in different solvents and its utilization in protein immobilization, tissue engineering, drug delivery, and inorganic reaction template. Therefore, based on recent studies, this review illustrated considerable progresses with addressing medicinal properties involving antimicrobial and biocompatibility of nano-cellulose (NC) in the case of wound healing.

Phoenix dactylifera lignocellulosic biomass as precursor for nanostructure fabrication using integrated process

Agricultural residue is an inevitable byproduct of agricultural activities. Valorization of agricultural residue is necessary to avoid wastage. In this study, we developed an integrated process for fabricating various nanostructures using Phoenix dactylifera (PD) lignocellulosic biomass as a sustainable bioprecursor. The PD biomass was treated using a hydrothermal method at 180 °C to derive the liquid fraction containing carbon nanodots (CDs). After pretreatment, the biomass was treated with an alkali solution to disassociate the lignin and cellulose. The extracted lignin was dissolved in tetrahydrofuran and subjected to a dialysis process. During the dialysis process, the lignin transformed into lignin nanoparticles (LNPs). The cellulose-containing fraction was transformed into cellulose nanostructures (CNs) via a sequential process (bleaching and acid hydrolysis). The structural properties, morphological features, crystallinity, and thermal stability of the fabricated CDs, LNPs, and CNs were analyzed. The synthesized CDs and LNPs had a spherical shape with different sizes: 2–10 nm (CDs) and 100–900 nm (LNPs). The CNs exhibited a fibrillated architecture and were 5–10 nm wide and 400–700 nm long. A biocompatibility analysis indicated that the synthesized nanostructures are nontoxic to human mesenchymal stem cells at concentrations of up to 200 μg/mL. This process is suitable for synthesizing nanostructures using different types of plant lignocellulosic biomass.

Heterologous co-expression of two β-glucanases and a cellobiose phosphorylase resulted in a significant increase in the cellulolytic activity of the Caldicellulosiruptor bescii exoproteome.

The ability to deconstruct plant biomass without conventional pretreatment has made members of the genus Caldicellulosiruptor the target of investigation for the consolidated processing of plant lignocellulosic biomass to biofuels and bioproducts. To investigate the synergy of enzymes involved and to further improve the ability of C. bescii to degrade cellulose, we introduced CAZymes that act synergistically with the C. bescii exoproteome in vivo and in vitro. We recently demonstrated that the Acidothermus cellulolyticus E1 endo-1,4-β-D-glucanase (GH5) with a family 2 carbohydrate-binding module (CBM) increased the activity of C. bescii exoproteome on biomass, presumably acting in concert with CelA. The β-glucanase, GuxA, from A. cellulolyticus is a multi-domain enzyme with strong processive exoglucanase activity, and the cellobiose phosphorylase from Thermotoga maritima catalyzes cellulose degradation acting synergistically with cellobiohydrolases and endoglucanases. We identified new chromosomal insertion sites to co-express these enzymes and the resulting strain showed a significant increase in the enzymatic activity of the exoproteome.

A heterodimeric glutathione S-transferase that stereospecifically breaks lignin's β(R)-aryl ether bond reveals the diversity of bacterial β-etherases

Lignin is a heterogeneous polymer of aromatic subunits that is a major component of lignocellulosic plant biomass. Understanding how microorganisms deconstruct lignin is important for understanding the global carbon cycle and could aid in developing systems for processing plant biomass into valuable commodities. Sphingomonad bacteria use stereospecific glutathione S-transferases (GSTs) called β-etherases to cleave the β-aryl ether (β-O-4) bond, the most common bond between aromatic subunits in lignin. Previously characterized bacterial β-etherases are homodimers that fall into two distinct GST subclasses: LigE homologues, which cleave the β(R) stereoisomer of the bond, and LigF homologues, which cleave the β(S) stereoisomer. Here, we report on a heterodimeric β-etherase (BaeAB) from the sphingomonad Novosphingobium aromaticivorans that stereospecifically cleaves the β(R)-aryl ether bond of the di-aromatic compound β-(2-methoxyphenoxy)-γ-hydroxypropiovanillone (MPHPV). BaeAB's subunits are phylogenetically distinct from each other and from other β-etherases, although they are evolutionarily related to LigF, despite the fact that BaeAB and LigF cleave different β-aryl ether bond stereoisomers. We identify amino acid residues in BaeAB's BaeA subunit important for substrate binding and catalysis, including an asparagine that is proposed to activate the GSH cofactor. We also show that BaeAB homologues from other sphingomonads can cleave β(R)-MPHPV and that they may be as common in bacteria as LigE homologues. Our results suggest that the ability to cleave the β-aryl ether bond arose independently at least twice in GSTs and that BaeAB homologues may be important for cleaving the β(R)-aryl ether bonds of lignin-derived oligomers in nature.

Engineer multi-functional cellulase/xylanase/β-glucosidase with improved efficacy to degrade rice straw

Plant lignocellulosic biomass can be enzymatically degraded and converted into biofuel. We have studied a bifunctional cellulase/xylanase from thermophilic Clostridium thermocellum (CtCel5E), which can digest two major plant cell wall polysaccharides, cellulose (a polymer of glucose) and xylan (a major hemicellulose composed of xylose), into disaccharides cellobiose and xylobiose, respectively. In the study presented here, the bifunctional CtCel5E was fused with a β-glucosidase from Clostridium cellulovorans (CcBglA) to convert cellulose into glucose with improved activities toward carboxymethyl cellulose (CMC), regenerated amorphous cellulose (RAC), and alkali-treated rice straw as compared to the mixture of two individual enzymes by eliminating accumulation of cellobiose intermediate. The engineered CtCel5E-CcBglA was secreted from yeast to degrade alkali-treated rice straw into glucose that sustained cell growth. Our study provides a strategy of consolidating enzymes for more efficient biomass processing in vitro and by yeast cells.

Growth studies of Leuconostoc mesenteroides on corn biomass based substrates

Leuconostoc mesenteroides is a well-known industrial producer of extracellular bacterial polysaccharide dextran. The main raw material for industrial cultivation of these microorganisms is molasses. Sucrose of molasses is the main source of carbon and energy for bacteria, as well as a precursor of dextran biosynthesis. It is known that the bacteria of the genus Leuconostoc are heterofermentative microorganisms and are capable to utilize various carbohydrates. The purpose of this study was to evaluate the of the corn biomass based mediumon the yield of bacterial biomass and the concentration of the extracellular polysaccharide.The hydrolyzate is a product of chemo-enzymatic treatment of plant lignocellulosic corn biomass and contains such monosaccharides as glucose, arabinose and xylose. The parameters of the composition of the medium were the concentration of hydrolyzate carbohydrates in the form of reducing substances (RS), the concentration of nitrogen (protein and non-protein), sucrose and the presence of minerals. The parameters of the cultivation process are pH and temperature. According to experimental data, the maximum yield of bacterial biomass (9.3 and 8.5%) were achieved on combined media with a sucrose content of 0.2% and a sodium salt (sodium phosphate), respectively. The highest specific growth rates of microorganisms (0.0840 and 0.0746) are demonstrated on a combined medium with a content of 0.15 and 0.3% inorganic nitrogen, respectively. The highest concentrations of extracellular polysaccharides (0.840 and 0.583%) are formed when magnesium and manganese salts are present in the nutrient medium. The optimal pH level for effective growth s is about 7, and the temperature in the range of 30-35 ° C.

Molecular-level driving forces in lignocellulosic biomass deconstruction for bioenergy

The plant cell wall biopolymers lignin, cellulose and hemicellulose are potential renewable sources of clean biofuels and high-value chemicals. However, the complex 3D structure of lignocellulosic biomass is recalcitrant to deconstruction. Major efforts to overcome this recalcitrance have involved pretreating biomass before catalytic processing. This Perspective describes recent work aimed at elucidating the molecular-level physical phenomena that drive biomass assembly. These are at play in commonly employed aqueous-based and thermochemical pretreatments. Several key processes have been found to be driven by biomass solvation thermodynamics, an understanding of which therefore facilitates the rational improvement of methods aimed at the complete solubilization and fractionation of the major biomass components. This Perspective describes the physical molecular driving forces that stabilize native lignocellulosic plant biomass structures and govern thermochemical biomass pretreatments. Understanding these driving forces can help us to design efficient methods for deconstructing biomass into biofuels and other bioproducts.

Paenibacillus polymyxa ND25: candidate genome for lignocellulosic biomass utilization.

Genome sequence of Paenibacillus polymyxa ND25 isolated from cow rumen is reported for being a potential candidate in hydrolysis of lignocellulosic plant biomass. Draft genome sequence generated 5.73Mb data containing 4922 putative protein coding genes, of which 140 are annotated for glycoside hydrolases. P. polymyxa ND25 strain comprises diverse lignocellulolytic components, especially 12 cellulase along with 23 hemicellulases and 11 esterases, signifying its potential for lignocellulose hydrolysis. Subsequent enzyme assay exhibited the potential of strain to produce 0.49, 0.24 and 0.44U/ml U/ml of endoglucanase, exoglucanase and β-glucosidase, respectively, utilizing sugarcane bagasse as the sole carbon source. This study signifies the efficient application of P. polymyxa ND25 for facilitating plant-biomass utilization.

Secreted protein extract analyses present the plant pathogen Alternaria alternata as a suitable industrial enzyme toolbox

Lignocellulosic plant biomass is the most abundant carbon source in the planet, which makes it a potential substrate for biorefinery. It consists of polysaccharides and other molecules with applications in pharmaceutical, food and feed, cosmetics, paper and textile industries. The exploitation of these resources requires the hydrolysis of the plant cell wall, which is a complex process. Aiming to discover novel fungal natural isolates with lignocellulolytic capacities, a screening for feruloyl esterase activity was performed in samples taken from different metal surfaces. An extracellular enzyme extract from the most promising candidate, the natural isolate Alternaria alternata PDA1, was analyzed. The feruloyl esterase activity of the enzyme extract was characterized, determining the pH and temperature optima (pH 5.0 and 55–60 °C, respectively), thermal stability and kinetic parameters, among others. Proteomic analyses derived from two-dimensional gels allowed the identification and classification of 97 protein spots from the extracellular proteome. Most of the identified proteins belonged to the carbohydrates metabolism group, particularly plant cell wall degradation. Enzymatic activities of the identified proteins (β-glucosidase, cellobiohydrolase, endoglucanase, β-xylosidase and xylanase) of the extract were also measured. These findings confirm A. alternata PDA1 as a promising lignocellulolytic enzyme producer. SignificanceAlthough plant biomass is an abundant material that can be potentially utilized by several industries, the effective hydrolysis of the recalcitrant plant cell wall is not a straightforward process. As this hydrolysis occurs in nature relying almost solely on microbial enzymatic systems, it is reasonable to infer that further studies on lignocellulolytic enzymes will discover new sustainable industrial solutions. The results included in this paper provide a promising fungal candidate for biotechnological processes to obtain added value from plant byproducts and analogous substrates. Moreover, the proteomic analysis of the secretome of a natural isolate of Alternaria sp. grown in the presence of one of the most used vegetal substrates on the biofuels industry (sugar beet pulp) sheds light on the extracellular enzymatic machinery of this fungal plant pathogen, and can be potentially applied to developing new industrial enzymatic tools. This work is, to our knowledge, the first to analyze in depth the secreted enzyme extract of the plant pathogen Alternaria when grown on a lignocellulosic substrate, identifying its proteins by means of MALDI-TOF/TOF mass spectrometry and characterizing its feruloyl esterase, cellulase and xylanolytic activities.

Evaluation of gastrointestinal bacterial population for the production of holocellulose enzymes for biomass deconstruction.

The gastrointestinal (GI) habitat of ruminant and non-ruminant animals sustains a vast ensemble of microbes that are capable of utilizing lignocellulosic plant biomass. In this study, an indigenous swine (Zovawk) and a domesticated goat (Black Bengal) were investigated to isolate bacteria having plant biomass degrading enzymes. After screening and enzymatic quantification of eighty-one obtained bacterial isolates, Serratia rubidaea strain DBT4 and Aneurinibacillus aneurinilyticus strain DBT87 were revealed as the most potent strains, showing both cellulase and xylanase production. A biomass utilization study showed that submerged fermentation (SmF) of D2 (alkaline pretreated pulpy biomass) using strain DBT4 resulted in the most efficient biomass deconstruction with maximum xylanase (11.98 U/mL) and FPase (0.5 U/mL) activities (55°C, pH 8). The present study demonstrated that bacterial strains residing in the gastrointestinal region of non-ruminant swine are a promising source for lignocellulose degrading microorganisms that could be used for biomass conversion.

Boosting LPMO-driven lignocellulose degradation by polyphenol oxidase-activated lignin building blocks

BackgroundMany fungi boost the deconstruction of lignocellulosic plant biomass via oxidation using lytic polysaccharide monooxygenases (LPMOs). The application of LPMOs is expected to contribute to ecologically friendly conversion of biomass into fuels and chemicals. Moreover, applications of LPMO-modified cellulose-based products may be envisaged within the food or material industry.ResultsHere, we show an up to 75-fold improvement in LPMO-driven cellulose degradation using polyphenol oxidase-activated lignin building blocks. This concerted enzymatic process involves the initial conversion of monophenols into diphenols by the polyphenol oxidase MtPPO7 from Myceliophthora thermophila C1 and the subsequent oxidation of cellulose by MtLPMO9B. Interestingly, MtPPO7 shows preference towards lignin-derived methoxylated monophenols. Sequence analysis of genomes of 336 Ascomycota and 208 Basidiomycota reveals a high correlation between MtPPO7 and AA9 LPMO genes.ConclusionsThe activity towards methoxylated phenolic compounds distinguishes MtPPO7 from well-known PPOs, such as tyrosinases, and ensures that MtPPO7 is an excellent redox partner of LPMOs. The correlation between MtPPO7 and AA9 LPMO genes is indicative for the importance of the coupled action of different monooxygenases in the concerted degradation of lignocellulosic biomass. These results will contribute to a better understanding in both lignin deconstruction and enzymatic lignocellulose oxidation and potentially improve the exploration of eco-friendly routes for biomass utilization in a circular economy.

The low affinity glucose transporter HxtB is also involved in glucose signalling and metabolism in Aspergillus nidulans

One of the drawbacks during second-generation biofuel production from plant lignocellulosic biomass is the accumulation of glucose, the preferred carbon source of microorganisms, which causes the repression of hydrolytic enzyme secretion by industrially relevant filamentous fungi. Glucose sensing, subsequent transport and cellular signalling pathways have been barely elucidated in these organisms. This study therefore characterized the transcriptional response of the filamentous fungus Aspergillus nidulans to the presence of high and low glucose concentrations under continuous chemostat cultivation with the aim to identify novel factors involved in glucose sensing and signalling. Several transcription factor- and transporter-encoding genes were identified as being differentially regulated, including the previously characterized glucose and xylose transporter HxtB. HxtB was confirmed to be a low affinity glucose transporter, localizing to the plasma membrane under low- and high-glucose conditions. Furthermore, HxtB was shown to be involved in conidiation-related processes and may play a role in downstream glucose signalling. A gene predicted to encode the protein kinase PskA was also identified as being important for glucose metabolism. This study identified several proteins with predicted roles in glucose metabolic processes and provides a foundation for further investigation into the response of biotechnologically important filamentous fungi to glucose.

Screening of rice mutants with improved saccharification efficiency results in the identification of CONSTITUTIVE PHOTOMORPHOGENIC 1 and GOLD HULL AND INTERNODE 1.

The screening of rice mutants with improved cellulose to glucose saccharification efficiency (SE) identifies reduced xylan and/or ferulic acid, and a qualitative change of lignin to impact SE. To ensure the availability of sustainable energy, considerable effort is underway to utilize lignocellulosic plant biomass as feedstock for the production of biofuels. However, the high cost of degrading plant cell wall components to fermentable sugars (saccharification) has been problematic. One way to overcome this barrier is to develop plants possessing cell walls that are amenable to saccharification. In this study, we aimed to identify new molecular factors that influence saccharification efficiency (SE) in rice. By screening 22 rice mutants, we identified two lines, 122 and 108, with improved SE. Reduced xylan and ferulic acid within the cell wall of line 122 were probable reasons of improved SE. Line 108 showed reduced levels of thioglycolic-released lignin; however, the amount of Klason lignin was comparable to the wild-type, indicating that structural changes had occurred in the 108 lignin polymer which resulted in improved SE. Positional cloning revealed that the genes responsible for improved SE in 122 and 108 were rice CONSTITUTIVE PHOTOMORPHOGENIC 1 (OsCOP1) and GOLD HULL AND INTERNODE 1 (GH1), respectively, which have not been previously reported to influence SE. The screening of mutants for improved SE is an efficient approach to identify novel genes that affect SE, which is relevant in the development of crops as biofuel sources.

Folding of xylan onto cellulose fibrils in plant cell walls revealed by solid-state NMR

Exploitation of plant lignocellulosic biomass is hampered by our ignorance of the molecular basis for its properties such as strength and digestibility. Xylan, the most prevalent non-cellulosic polysaccharide, binds to cellulose microfibrils. The nature of this interaction remains unclear, despite its importance. Here we show that the majority of xylan, which forms a threefold helical screw in solution, flattens into a twofold helical screw ribbon to bind intimately to cellulose microfibrils in the cell wall. 13C solid-state magic-angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy, supported by in silico predictions of chemical shifts, shows both two- and threefold screw xylan conformations are present in fresh Arabidopsis stems. The twofold screw xylan is spatially close to cellulose, and has similar rigidity to the cellulose microfibrils, but reverts to the threefold screw conformation in the cellulose-deficient irx3 mutant. The discovery that induced polysaccharide conformation underlies cell wall assembly provides new principles to understand biomass properties.