Recovery of Polyphenols from Grape Pomace Using Polyethylene Glycol (PEG)-Grafted Silica Particles and PEG-Assisted Cosolvent Elution.

Adsorption on a functionalized surface can be an effective way of purifying polyphenols from complex plant extracts. Polymeric resins that rely on hydrophobic interactions suffer from low selectivity, weak affinity towards polyphenols, and lack tunability therefore making the purification of polyphenols less efficient. In this study, a purification process for the recovery of polyphenols from grape pomace extract was successfully developed using hydrogen bonding affinity ligands grafted on silica particles and PEG-assisted elution solvents. Bare silica (SiO2) and polyethylene glycol (mPEG)-grafted silica microparticles with molecular weights of 2000 and 5000 were tested to determine their polyphenol binding and release characteristics. Functionalizing the surface of bare silica with mPEG ligands increased the adsorption capacity by 7.1- and 11.4-fold for mPEG-2000 and mPEG-5000 compared to bare silica particles, respectively. This was likely due to the introduction of more polyphenol binding sites with mPEG functionalization. Altering the molecular weight (MW) of mPEG grafted on silica surfaces provided tunability in the adsorption capacity. A complete recovery of polyphenols (~99.9%) from mPEG-grafted silica particles was achieved by utilizing PEG-ethanol or PEG-water cosolvent systems. Recovered polyphenols showed up to ~12-fold antioxidant activity compared to grape pomace extract. This study demonstrates that mPEG-grafted silica particles and elution of polyphenols with PEG cosolvents can potentially be used for large-scale purification of polyphenols from complex plant extracts and simplify the use of polyphenols, as PEG facilitates remarkable solvation and is an ideal medium for the final formulation of polyphenols.

Importance of "weak-base" poplar wastes to process performance and methane yield in solid-state anaerobic digestion.

Failure of methane yield is common for anaerobic digestion (AD) of "weak-acid/acid" wastes alone. In order to verify the importance of pH of materials on the process performance and the methane yield, the "weak-base" wastes-poplar wastes (PW) were used as substrate of solid-state AD (SS-AD). The results show that PW could be used for efficient methane yield after NaOH treatment, the total methane yield was 81.1 L/kg volatile solids (VS). PW also could be used for anaerobic co-digestion with high-pH cattle slurry (CM). For the group with NaOH pretreatment, time used for reaching stable state was 2 days earlier than that of the group without NaOH pretreatment. The maximal methane yield of 98.2 L/kg VS was obtained on conditions of 1:1 of PW-to-CM (P/C) ratio and NaOH pretreatment, which was 21.1% (p < 0.05) higher than that of PW. The maximal reductions of total solids (TS), VS, cellulose and hemicellulose were 51.3%, 57.5%, 46.0% and 47.0%, respectively, which were associated with the maximal methane yield. The results indicate that PW could be alone used for efficient SS-AD for methane yield after NaOH treatment.

Screening of ligninolytic fungi for biological pretreatment of lignocellulosic biomass.

To identify white rot fungi with high potential in biological pretreatment of lignocellulosic biomass, preliminary screening was carried out on plates by testing different strains for their ability to oxidize guaiacol and decolorize the dyes azure B and Poly R-478. Of the 86 strains screened, 16 were selected for secondary screening for their ligninolytic ability; however, low manganese peroxidase activity and no lignin peroxidase activity were detected. Strain BBEL0970 proved to be the most efficient in laccase production and was subsequently identified as Trametes versicolor by analysis of the ribosomal DNA internal transcribed spacer gene sequence. In combining laccase production with biological pretreatment, the replacement of glucose with barley straw significantly improved the laccase activity by up to 10.3 U/mL, which provided evidence toward potential utilization of barley straw in laccase production by BBEL0970. Simultaneously, comparison by thermogravimetric analysis of the untreated and pretreated barley straw in liquid fermentation of laccase also demonstrated the high potential of BBEL0970 in biological pretreatment of lignocellulosic biomass. This work sheds light on further exploration on the integrated process of low-cost laccase production and efficient biological pretreatment of barley straw by T. versicolor BBEL0970.

Mutagenesis and evaluation of cellulase properties and cellulose hydrolysis of Talaromyces piceus.

A fungal species with a high yield of ß-glucosidase was isolated and identified as Talaromyces piceus 9-3 (anamorph: Penicillium piceum) by morphological and molecular characterization. Through dimethyl sulphate mutagenesis, the cellulase over-producing strain T. piceus H16 was obtained. The FPase activity and ß-glucosidase activity of T. piceus H16 were 5.83 and 53.12 IU ml(-1) respectively--a 5.34- and 4.43-times improvement from the parent strain T. piceus 9-3. The optimum pH and temperature for enzyme activity were pH 5.0 and 50 °C for FPase activity and pH 5.0 and 55 °C for ß-glucosidase activity, respectively. The cellulase were quite stable at 37 °C, only losing <10% of their initial activity after 24 h of incubation. Hydrolysis analysis results showed that a highly efficient synergistic effect was achieved by combining cellulase from T. piceus H16 with that from Trichoderma reesei RUT C30 on hydrolyzing different substrates due to the high ß-glucosidase activity of T. piceus H16. These data suggest that T. piceus H16 can be used as a potential cellulase producer with good prospects.

Effect of earlier unfolded protein response and efficient protein disposal system on cellulase production in Rut C30.

Trichoderma reesei (T. reesei) has been widely used in production of cellulolytic enzymes and heterologous proteins because of its high secretion capacity. The lack of knowledge on protein secretion mechanisms, however, still hinders rational improvement on cellulase production. The transcript levels of cellulases and components involved in post-transcriptional procedures were compared in this study between two mutants, QM9414 and Rut C30 for evaluating the effects of modification and secretion upon cellulase production. The results showed that cellulase induction by cellulose drastically up-regulated expressions of the sensor of unfolded protein, chaperone and folding-assisted enzymes in endoplasmic reticulum and resulted in unfolded protein response (UPR) and low-grade increase in secretory transporters' expression similar to that of chemical treatment. Rut C30 demonstrated earlier and more sustainable expressions of elements involved in UPR and lower amount of cellular retained cellulase compared to QM9414, indicating that Rut C30 had hypercellulolytic property partially for its earlier and enhanced UPR to more efficiently dispose of protein. Modifying post-translational peptides and enhancing protein flux to avoid protein accumulation during cellulase production may be a feasible approach for strain improvement.

Nature-inspired pretreatment of lignocellulose - Perspective and development.

As sustainability gains increasing importance in addition to cost-effectiveness as a criterion for evaluating engineering systems and practices, biological processes for lignocellulose pretreatment have attracted growing attention. Biological systems such as white and brown rot fungi and wood-consuming insects offer fascinating examples of processes and systems built by nature to effectively deconstruct plant cell walls under environmentally benign and energy-conservative environments. Research in the last decade has resulted in new knowledge that advanced the understanding of these systems, provided additional insights into these systems' functional mechanisms, and demonstrated various applications of these processes. The new knowledge and insights enable the adoption of a nature-inspired strategy aiming at developing technologies that are informed by the biological systems but superior to them by overcoming the inherent weakness of the natural systems. This review discusses the nature-inspired perspective and summarizes related advancements, including the evolution from biological systems to nature-inspired processes, the features of biological pretreatment mechanisms, the development of nature-inspired pretreatment processes, and future perspective. This work aims to highlight a different strategy in the research and development of novel lignocellulose pretreatment processes and offer some food for thought.

Biodegradation of hardwood lignocellulosics by the western poplar clearwing borer, Paranthrene robiniae (Hy. Edwards).

Lignin in plant cell wall is a source of useful chemicals and also the major barrier for saccharification of lignocellulosic biomass for producing biofuel and bioproducts. Enzymatic lignin degradation/modification process could bypass the need for chemical pretreatment and thereby facilitate bioprocess consolidation. Herein, we reveal our new discovery in elucidating the process of hardwood lignin modification/degradation by clearwing borer, Paranthrene robiniae . The wood-boring clearwing borer, P. robiniae , effectively tunnels hardwood structures during the larval stage; its digestion products from wood components, however, has not yet been investigated. A series of analysis conducted in this study on tunnel walls and frass produced provided evidence of structural alterations and lignin degradation during such hardwood digestion process. The analysis included solid state (13)C cross-polarization magic angle spinning (CP/MAS) nuclear magnetic resonance (NMR) spectroscopy, attenuated total reflectance Fourier transform infrared (ATR-FTIR), pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS), and thermogravimetric (TG) analysis; the results strongly suggest that the structural alteration of lignin primarily involved a preferential degradation of syringyl units accompanied by oxidation on the side chains of lignin guaiacyl moieties. This study also further indicated that unlike the wood-feeding termite the clearwing borer does not target cellulose as an energy source, and thus its lignin degradation ability should provide potential information on how to disassemble and utilize hardwood lignin. Overall, this biological model with an efficient lignin disruption system will provide the new insight into novel enzyme system required for effective plant cell wall disintegration for enhanced cellulose accessibility by enzymes and production of value-added lignin derived products.

Potential of biofilm-based biofuel production.

Biofilm technology has been extensively applied to wastewater treatment, but its potential application in biofuel production has not been explored. Current technologies of converting lignocellulose materials to biofuel are hampered by costly processing steps in pretreatment, saccharification, and product recovery. Biofilms may have a potential to improve efficiency of these processes. Advantages of biofilms include concentration of cell-associated hydrolytic enzymes at the biofilm-substrate interface to increase reaction rates, a layered microbial structure in which multiple species may sequentially convert complex substrates and coferment hexose and pentose as hydrolysates diffuse outward, and the possibility of fungal-bacterial symbioses that allow simultaneous delignification and saccharification. More importantly, the confined microenvironment within a biofilm selectively rewards cells with better phenotypes conferred from intercellular gene or signal exchange, a process which is absent in suspended cultures. The immobilized property of biofilm, especially when affixed to a membrane, simplifies the separation of biofuel from its producer and promotes retention of biomass for continued reaction in the fermenter. Highly consolidated bioprocessing, including delignification, saccharification, fermentation, and separation in a single reactor, may be possible through the application of biofilm technology. To date, solid-state fermentation is the only biofuel process to which the advantages of biofilms have been applied, even though it has received limited attention and improvements. The transfer of biofilm technology from environmental engineering has the potential to spur great innovations in the optimization of biofuel production.

Duckweed (Lemna minor) is a novel natural inducer of cellulase production in Trichoderma reesei.

An inducer is crucial for cellulase production. In this study, duckweed was used as an inducer of cellulase production by Trichoderma reesei RUT C30. In a reaction induced by 50 g/L duckweed in shake flasks, the filter-paper activity (FPA) reached 6.5 FPU/mL, a value comparable to that induced by avicel. The enzyme-hydrolysis rate induced by steam-exploded corn stalk was 54.2%, representing a 28% improvement over that induced by avicel. The duckweed starch was hydrolyzed to glucose, which was subsequently used for biomass accumulation during the fermentation process. Furthermore, to optimize the control of the fermentation process, a combined substrate of avicel and duckweed was used to induce cellulase production by T. reesei RUT C30. The cellulase production and hydrolysis rates of the combined substrate, compared with avicel alone, were 39.6% and 36.7% higher, respectively. The results of this study suggest that duckweed is a good inducer of cellulase production in T. reesei, and it might aid in decreasing the cost of lignocellulosic materials hydrolysis.

Kinetic modeling of enzymatic hydrolysis of cellulose in differently pretreated fibers from dairy manure.

A kinetic model incorporating dynamic adsorption, enzymatic hydrolysis, and product inhibition was developed for enzymatic hydrolysis of differently pretreated fibers from a nitrogen-rich lignocellulosic material-dairy manure. The effects of manure proteins on the enzyme adsorption profile during hydrolysis have been discussed. Enzyme activity, instead of protein concentration, was used to describe the enzymatic hydrolysis in order to avoid the effect of manure protein on enzyme protein analysis. Dynamic enzyme adsorption was modeled based on a Langmiur-type isotherm. A first-order reaction was applied to model the hydrolysis with consideration being given for the product inhibition. The model satisfactorily predicted the behaviors of enzyme adsorption, hydrolysis, and product inhibition for all five sample manure fibers. The reaction conditions were the substrate concentrations of 10-50 g/L, enzyme loadings of 7-150 FPU/g total substrate, and the reaction temperature of 50 degrees C.

The white-rot fungus Phanerochaete chrysosporium: conditions for the production of lignin-degrading enzymes.

Investigating optimal conditions for lignin-degrading peroxidases production by Phanerochaete chrysosporium (P. chrysosporium) has been a topic for numerous researches. The capability of P. chrysosporium for producing lignin peroxidases (LiPs) and manganese peroxidases (MnPs) makes it a model organism of lignin-degrading enzymes production. Focusing on compiling and identifying the factors that affect LiP and MnP production by P. chrysosporium, this critical review summarized the main findings of about 200 related research articles. The major difficulty in using this organism for enzyme production is the instability of its productivity. This is largely due to the poor understanding of the regulatory mechanisms of P. chrysosporium responding to different nutrient sources in the culture medium, such as metal elements, detergents, lignin materials, etc. In addition to presenting the major conclusions and gaps of the current knowledge on lignin-degrading peroxidases production by P. chrysosporium, this review has also suggested further work, such as correlating the overexpression of the intra and extracellular proteins to the nutrients and other culture conditions to discover the regulatory cascade in the lignin-degrading peroxidases production process, which may contribute to the creation of improved P. chrysosporium strains leading to stable enzyme production.

Co-production of fumaric acid and chitin from a nitrogen-rich lignocellulosic material - dairy manure - using a pelletized filamentous fungus Rhizopus oryzae ATCC 20344.

Fumaric acid is widely used as a food additive for flavor and preservation. Rhizopus oryzae ATCC 20344 is a fungus known for good fumaric acid production. It also has been reported that the fungal biomass has high chitin content. This study investigated the possibility of producing both fumaric acid and chitin via R. oryzae fermentation of dairy manure. Co-production of valuable bio-based chemicals such as fumaric acid and chitin could make the utilization of manure more efficient and more profitable. A three step fermentation process was developed which effectively utilized the nitrogen as well as the carbohydrate sources within the manure. These steps were: the culturing of pellet seed; biomass cultivation on liquid manure to produce both biomass and chitin; and fumaric acid production on the hydrolysate from the manure fiber. Under the identified optimal conditions, the fermentation system had a fumaric acid yield of 31%, and a biomass concentration of 11.5 g/L that contained 0.21 g chitin/g biomass.

Studying the effects of reaction conditions on components of dairy manure and cellulose accumulation using dilute acid treatment.

The objectives of this study were to statistically study the effects of reaction conditions of temperature, acid concentration, and reaction time on manure components of cellulose, hemicellulose, lignin, and nitrogen during dilute acid treatment of dairy manure; and to further optimize the accumulation of cellulose for later enzymatic conversion to glucose. A 2(3) full factorial design was adopted to investigate the effects of the reaction conditions on each individual component and later followed by a 3-factor central composite design which was used to obtain the optimal conditions for cellulose accumulation. The results indicated that acid was the most important factor for changes of all the components. The results also showed that two other individual factors, reaction time and temperature, as well as the interactions among all three factors had significant influences on the changes. In addition, the optimal conditions for cellulose accumulation were 2.8h reaction time, 140 degrees C reaction temperature, and 1.0% acid concentration. Under these conditions cellulose content reached 31.0% while hemicellulose, lignin and nitrogen content were 3.2%, 20.8% and 2.4%, respectively.

Acid hydrolysis of fibers from dairy manure.

Concentrated acid hydrolysis of lignocellulosic materials is a conventional treatment process for the production of mono-sugars. However, this method has been proved ineffective and undesirable for the treatment of dairy manure due to the high nitrogen content of dairy manure and the environmental issues caused by the use of highly concentrated acid solution. In an effort to overcome these barriers, a modified acid hydrolysis process with short reaction time was introduced that involved a nitrogen-removing pretreatment followed by decrystallization with concentrated acid and then hydrolysis using dilute acid. The effects of nitrogen, acid concentration, reaction time, and temperature were investigated. A pretreated manure with a low nitrogen content of 1.3% was used as the substrate. The results indicated that the optimal conditions for fiber decrystallization were 75% acid concentration, 3:5 sample to acid ratio (weight basis), and 30 min of reaction time; while the optimal conditions for acid hydrolysis were 12.5% acid and 10% dry sample at 135 degrees C for 10 min. These conditions produced 26 g/L glucose at a yield of 84% and 11 g/L hemicellulose-sugars at a yield of 80%.

Thermal behavior and kinetic study for catalytic co-pyrolysis of biomass with plastics.

The present study aims to investigate the thermal decomposition behaviors and kinetics of biomass (cellulose/Douglas fir sawdust) and plastics (LDPE) in a non-catalytic and catalytic co-pyrolysis over ZSM-5 catalyst by using a thermogravimetric analyzer (TGA). It was found that there was a positive synergistic interaction between biomass and plastics according to the difference of weight loss (ΔW), which could decrease the formation of solid residue at the end of the experiment. The first order reaction model well fitted for both non-catalytic and catalytic co-pyrolysis of biomass with plastics. The activation energy (E) of Cellulose-LDPE-Catalyst and DF-LDPE-Catalyst are only 89.51 and 54.51kJ/mol, respectively. The kinetics analysis showed that adding catalyst doesn't change the decomposition mechanism. As a result, the kinetic study on catalytic co-pyrolysis of biomass with plastics was suggested that the catalytic co-pyrolysis is a promising technique that can significantly reduce the energy input.

Optimizing carbon efficiency of jet fuel range alkanes from cellulose co-fed with polyethylene via catalytically combined processes.

Enhanced carbon yields of renewable alkanes for jet fuels were obtained through the catalytic microwave-induced co-pyrolysis and hydrogenation process. The well-promoted ZSM-5 catalyst had high selectivity toward C8-C16 aromatic hydrocarbons. The raw organics with improved carbon yield (∼44%) were more principally lumped in the jet fuel range at the catalytic temperature of 375°C with the LDPE to cellulose (representing waste plastics to lignocellulose) mass ratio of 0.75. It was also observed that the four species of raw organics from the catalytic microwave co-pyrolysis were almost completely converted into saturated hydrocarbons; the hydrogenation process was conducted in the n-heptane medium by using home-made Raney Ni catalyst under a low-severity condition. The overall carbon yield (with regards to co-reactants of cellulose and LDPE) of hydrogenated organics that mostly match jet fuels was sustainably enhanced to above 39%. Meanwhile, ∼90% selectivity toward jet fuel range alkanes was attained.

Effects of hemicellulose and lignin on enzymatic hydrolysis of cellulose from dairy manure.

This study focused on the effect of hemicellulose and lignin on enzymatic hydrolysis of dairy manure and hydrolysis process optimization to improve sugar yield. It was found that hemicellulose and lignin in dairy manure, similar to their role in other lignocellulosic material, were major resistive factors to enzymatic hydrolysis and that the removal of either of them, or for best performance, both of them, improved the enzymatic hydrolysis of manure cellulose. This result combined with scanning electron microscope (SEM) pictures further proved that the accessibility of cellulose to cellulase was the most important feature to the hydrolysis. Quantitatively, fed-batch enzymatic hydrolysis of fiber without lignin and hemicellulose had a high glucose yield of 52% with respect to the glucose concentration of 17 g/L at a total enzyme loading of 1300 FPU/L and reaction time of 160 h, which was better than corresponding batch enzymatic hydrolysis.

Structural characterization of lignin: a potential source of antioxidants guaiacol and 4-vinylguaiacol.

The structure of lignin obtained from the ozone and soaking aqueous ammonia pretreatment of wheat straw has been characterized utilizing chemical analytical methods in order to reveal its antioxidant characteristics, including attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS), pyrolysis/tetramethylammonium hydroxide-gas chromatography/mass spectrometry (Py/TMAH-GC/MS), gel permeation chromatography (GPC), ultra violet-visible spectroscopy (UV-vis), and 1,1-diphenyl-2-picrylhydrazyl (DPPH) antioxidant evaluation assay. The results demonstrated that the isolated lignin is a ρ-hydroxyphenyl- guaiacyl-syringyl (H-G-S) lignin, with S/G ratio of 0.35 and significant amounts of phenol 2-methoxy (guaiacol) and phenol 2-methoxy-4-vinyl (4-vinylguaiacol). The Py-GC/MS and Py/TMAH-GC/MS pyrograms indicated that the major units in this lignin are derived from hydroxycinnamic acids. The GPC results revealed the molecular weight of the lignin was considerably low and also the FTIR analysis showed that the lignin possessed hydroxyl and methoxy functional groups; the factors led to the extracted lignin having a comparable antioxidant activity to that of currently used commercial antioxidants. The UV-vis and DPPH antioxidant assay results suggested a percentage of inhibition of the DPPH radicals in the following order: guaiacol (103.6 ± 1.36)>butylated hydroxytoluene (103.3 ± 1)>ferulic acid (102.6 ± 0.79)>pretreated lignin (86.9 ± 0.34).

Simulation of the ozone pretreatment of wheat straw.

Wheat straw is a potential feedstock in biorefinery for sugar production. However, the cellulose, which is the major source of sugar, is protected by lignin. Ozonolysis deconstructs the lignin and makes cellulose accessible to enzymatic digestion. In this study, the change in lignin concentration with different ozonolysis times (0, 1, 2, 3, 5, 7, 10, 15, 20, 30, 60min) was fit to two different kinetic models: one using the model developed by Garcia-Cubero et al. (2012) and another including an outer mass transfer barrier or "cuticle" region where ozone mass transport is reduced in proportion to the mass of unreacted insoluble lignin in the cuticle. The kinetic parameters of two mathematical models for predicting the soluble and insoluble lignin at different pretreatment time were determined. The results showed that parameters derived from the cuticle-based model provided a better fit to experimental results compared to a model without a cuticle layer.

Hydrolysis of animal manure lignocellulosics for reducing sugar production.

Converting animal manure into value-added products provides a potential alternative for treatment and disposal of such materials. Lignocellulosics are a major component of animal manure and represent an undeveloped bioresource. In this work, a process was developed for hydrolyzing manure lignocellulosics into fermentable sugars. When raw dairy manure was pre-treated with 3% sulfuric acid at 110 degrees C for 1 h, hemicellulose was completely degraded into mainly arabinose, galactose and xylose. The pretreated materials were then treated with cellulolytic enzymes, Celluclast-1.5L and Novozyme-188, to hydrolyze the cellulose. The optimal enzyme loadings were identified as 13 FPU cellulase/g substrate and 5 IU beta-glucosidase/g substrate. The optimal temperature and pH were determined to be 46 degrees C and 4.8, respectively. A substrate concentration of 50 g/l favored both glucose concentration (in hydrolysate) and glucose yield (based on per 100 g manure). It was also found that a reduced particle size of 590-mum resulted in a high glucose yield with further decreases in particle size not increasing the yield. For each particle size investigated, the addition of 2% tween-80 resulted in at least 20% improvement in glucose yield. The optimized hydrolysis process achieved a glucose yield of 11.32 g/100 g manure, which corresponded to about 40% cellulose conversion.