Fractionation of oil palm fronds using ethanol-assisted deep eutectic solvent: Influence of ethanol concentration on enhancing enzymatic saccharification and lignin ß-O-4 content.

Numerous fractionation methods have been developed in recent years for separating components such as cellulose, hemicellulose, and lignin from lignocellulosic biomass wastes. Deep eutectic solvents (DES) have recently been widely investigated as captivating green solvents for biomass fractionation. However, most acidic-based deep eutectic solvent fractionation produces condensed lignin with low ß-O-4 content. Besides, most DESs exhibit high viscosity, which results in poor mass transfer properties. This study aimed to address the challenges above by incorporating ethanol into the deep eutectic solvent at various concentrations (10-50 wt%) to fractionate oil palm fronds at a mild condition, i.e., 80 °C, 1 atm. Cellulose residues fractionated with ethanol-assisted deep eutectic solvent showed a maximum glucose yield of 85.8% when 20 wt% of ethanol was incorporated in the deep eutectic solvent, significantly higher than that achieved by pure DES (44.8%). Lignin extracted with ethanol-assisted deep eutectic solvent is lighter in color and higher in ß-O-4 contents (up to 44 ß-O-4 per 100 aromatic units) than pure DES-extracted lignin. Overall, this study has demonstrated that incorporating ethanol into deep eutectic solvents could enhance the applicability of deep eutectic solvents in the complete valorization of lignocellulosic biomass. Highly enzymatic digestible cellulose-rich solid and ß-O-4-rich lignin attained from the fractionation could serve as sustainable precursors for the production of biofuels.

Novel techniques for the mass production of nutritionally improved, fungus-treated lignocellulosic biomass for ruminant nutrition.

BACKGROUND: Laboratory-scale experiments have shown that treatment with selective lignin-degrading white-rot fungi improves the nutritional value and ruminal degradability of lignocellulosic biomass (LCB). However, the lack of effective field-applicable pasteurization methods has long been recognized as a major obstacle for scaling up the technique for fungal treatment of large quantities of LCB for animal feeding. In this study, wheat straw (an LCB substrate) was subjected to four field-applicable pasteurization methods - hot-water, formaldehyde fumigation, steam, and hydrated lime - and cultured with Pleurotus ostreatus grain spawn for 10, 20, and 30 days under solid-state fermentation. Samples of untreated, pasteurized but non-inoculated and fungus-treated straws were analyzed for chemical composition, aflatoxin B1 (AFB1 ), and in vitro dry matter digestibility (IVDMD), in vitro total gas (IVGP), methane (CH4 ), and volatile fatty acid (VFA) production. RESULTS: During the 30-day fungal treatment, steam and lime pasteurized straws had the greatest loss of lignin, resulting in marked improvements in crude protein (CP), IVDMD, IVGP, and total VFAs. Irrespective of the pasteurization method, the increase in IVDMD during fungal treatment was linearly (R2 = 0.77-0.92) related to lignin-loss in the substrate during fungal treatment. The CH4 production of the fungus-treated straw was not affected by the pasteurization methods. Aflatoxin B1 was within the safe level (<5 µg kg-1 ) in all pasteurized, fungus treated straws. CONCLUSION: Steam and lime were promising field-applicable pasteurization techniques to produce nutritionally improved fungus-treated wheat straw to feed ruminants. Lime pasteurization was more economical and did not require expensive energy inputs. © 2023 Society of Chemical Industry.

Deep Eutectic Solvents for Biotechnology Applications.

Deep eutectic solvents (DESs) are an alternative to traditional organic solvents and ionic liquids and meet the requirements of "green" chemistry. They are easy to prepare using low-cost constituents, are non-toxic and biodegradable. The review analyzes literature on the use of DES in various fields of biotechnology, provides data on the types of DESs, methods for their preparation, and properties. The main areas of using DESs in biotechnology include extraction of physiologically active substances from natural resources, pretreatment of lignocellulosic biomass to improve enzymatic hydrolysis of cellulose, production of bioplastics, as well as a reaction medium for biocatalytic reactions. The aim of this review is to summarize available information on the use of new solvents for biotechnological purposes.

Cryo-Induced Cellulose-Based Nanogel from Elaeis guineensis for Antibiotic Delivery Platform.

Cryo-induced hydrogel from cellulose is a new class of biomaterials for drug delivery, cell delivery, bone and skin tissue engineering for cell proliferation and regeneration applications. This research aimed to synthesize cryo-induced hydrogel from cellulose and carboxymethyl cellulose (CMC) produced from empty bunch's cell wall of Elaeis guineensis. First, the experiment was to produce cellulose-rich material using hot-compressed water extraction followed by alkaline delignification and bleaching with H2O2. The obtained bleached EFB cellulose was used as the substrate for CMC, and the optimal condition with the highest degree of carboxyl substitution (DS) of 0.75 was achieved when varying NaOH and monochloroacetic acid concentration as well as etherification temperature using fractional factorial design. For cryogelation study, hydrogels were synthesized from cellulose, CMC and beta-cyclodextrin (ß-CD) by dissolving cellulose-based matrix in a NaOH/urea system, and the cellulose (CEL) solution was frozen spontaneously at -40 °C followed by high speed mixing to loosen cellulose fibrils. Epichlorohydrin (ECH) and Polyethylene glycol diglycidyl ether (PEGDE) were used as a cross-linker. First, the ratio of cellulose and CMC with different amounts of ECH was investigated, and subsequently the proper ratio was further studied by adding different crosslinkers and matrices, i.e., CMC and ß-CD. From the result, the ECH crosslinked CMC-CEL (E-CMC-CEL) gel had the highest swelling properties of 5105% with the average pore size of lyophilized hydrogel of 300 µm. In addition, E-CMC-CEL gel had the highest loading and release capability of tetracycline in buffer solution at pH 7.4 and 3.2. At pH 7.4, tetracycline loading and release properties of E-CMC-CEL gel were 65.85 mg g-1 dry hydrogel and 46.48 mg g-1 dry hydrogel (70.6% cumulative release), respectively. However, at pH 3.2, the loading and release capabilities of Tetracycline were moderately lower at 16.25 mg g-1 dry hydrogel and 5.06 mg g-1 dry hydrogel, respectively. The findings presented that E-CMC-CEL hydrogel was a suitable material for antibiotic tetracycline drug carrying platform providing successful inhibitory effect on Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, respectively.

Agro-industrial wastes revalorization as feedstock: production of lignin-modifying enzymes extracts by solid-state fermentation using white rot fungi.

The purpose of the study was to evaluate the production of lignin-modifying enzyme extracts and delignified biomass from agro-industrial wastes using white rot fungi (Inonotus sp. Sp2, Stereum hirsutum Ru-104, Bjerkandera sp. BOS55, Pleurotus eryngii IJFM 169 and Phanerochaete chrysosporium BKM-F-1767). These were screened based on their adaptability and colonization ability on different substrates, as well as by the Laccase, Manganese peroxidase, and Lignin peroxidase enzymatic production. Native strains (Inonotus sp. Sp2 and S. hirsutum Ru-104) showed the highest growth kinetics under the solid-substrate fermentation conditions and the growth rate parameters of the kinetic logistic model for the different substrates were between 0.39-0.81 (1/d) and 0.42-0.83 (1/d), respectively; the determination coefficients were ≥0.99. Inonotus sp. Sp2 was subsequently cultured in static flasks to produce crude enzyme extracts, obtaining manganese peroxidase activity levels of 18.5 and 31.3 (U/g) when growing in corn cob husk and spent tea leaves, respectively. Besides, it was to establish that the best conditions for lignin-modifying enzymes production using corn cob husk are 70% of initial moisture and 2.12 mm of particle size; reaching after 30 incubation days a manganese peroxidase activity of 21 ± 6 (U/g) under these conditions; enzyme that showed a suitable thermostability.

Improving the Cellulose Enzymatic Digestibility of Sugarcane Bagasse by Atmospheric Acetic Acid Pretreatment and Peracetic Acid Post-Treatment.

Pretreatment of sugarcane bagasse (SCB) by aqueous acetic acid (AA), with the addition of sulfuric acid (SA) as a catalyst under mild condition (<110 °C), was investigated. A response surface methodology (central composite design) was employed to study the effects of temperature, AA concentration, time, and SA concentration, as well as their interactive effects, on several response variables. Kinetic modeling was further investigated for AA pretreatment using both Saeman's model and the Potential Degree of Reaction (PDR) model. It was found that Saeman's model showed a great deviation from the experimental results, while the PDR model fitted the experimental data very well, with determination coefficients of 0.95-0.99. However, poor enzymatic digestibility of the AA-pretreated substrates was observed, mainly due to the relatively low degree of delignification and acetylation of cellulose. Post-treatment of the pretreated cellulosic solid well improved the cellulose digestibly by further selectively removing 50-60% of the residual linin and acetyl group. The enzymatic polysaccharide conversion increased from <30% for AA-pretreatment to about 70% for PAA post-treatment.

Integral valorisation of walnut shells based on a three-step sequential delignification.

Walnut kernels represent no more than 50-60% of the total weight of the fruit, so the sum of walnut shells generated every year is immense. Nonetheless, these shells could be further valorised for the extraction of their main constituents following a biorefinery scheme. Hence, the objective of this work was an integral valorisation of walnut shells, which involved a sequential organosolv delignification (200 °C, 90 min, 70/30 v/v EtOH/H2O, LSR 6:1) and several posterior non-isothermal hydrothermal treatments (180, 195 and 210 °C, LSR 8:1). Moreover, the spent solids after the aforementioned treatments were evaluated as possible sources of cellulose nanocrystals. The results showed that the sequential organosolv delignifications presented relative lignin yields up to 60%, which leaded to lignins that just differed on their molecular weight distributions. The hydrothermal treatments were efficient for the removal of still present hemicelluloses (14.7-71.8%), and permitted a successful cellulose nanocrystal obtaining whereas the spent solid from the delignification stages did not. Thus, this study presented an innovative strategy for the integral valorisation of walnut shells.

Dual phase statistical optimization of biological pre-treatment of sugarcane bagasse with Pycnoporus coccineus MScMS1 for polyhydroxyalkanoates production.

Biological pre-treatment is the removal of recalcitrant lignin from lignocellulose through the action of lignin degrading organisms and/or their ligninolytic enzymes system. Despite numerous environmental benefits, biological pre-treatment has been side-lined due to its prolonged periods of fermentation, ascribed to the slow growth rate of lignin degrading organisms. Thus, the present work adopted a dual phase statistical optimization approach for the biological pre-treatment of sugarcane bagasse, with Pycnoporus coccineus MScMS1, using Taguchi Orthogonal Array, in conjunction with Response Surface Methodology, to address this issue. Amplification of the organism's functioning resulted in an enhancement of sugar productivity and yield accompanied by a significant reduction in fermentation time. Optimized sugar concentration was approx. 18 g/L within 4 days of pre-treatment, with productivity of 4.5 g/(L.day). Substrate compositional analysis revealed significant (p < 0.05) reduction of lignin by 70% in the biologically pre-treated substrate, along with significantly (p < 0.05) higher quantities of water soluble components (35 ± 0.95 g) and cellulose content (33 ± 0.18 g), as compared to the untreated substrate. Appreciable levels of xylose, arabinose, glucose and galactose were detected in hydrolysates from biologically pre-treated bagasse. Furthermore, Bacillus megaterium Ti3, a potent polyhydroxyalkanoates (PHA) producer, was grown on these sugar-rich hydrolysates and generated 0.58 g/L PHA in 24 h of fermentation accompanied by 0.88 g/L dry cell weight and 65% PHA accumulation. These results were comparable with those from a glucose medium. Thus, the present study was successful in optimizing the biological pre-treatment of sugarcane bagasse and utilizing the resultant sugar-rich hydrolysates, as inexpensive and renewable raw materials, for PHA production.

Molecular Characteristics and Antioxidant Activity of Spruce (Picea abies) Hemicelluloses Isolated by Catalytic Oxidative Delignification.

Spruce (Piceaabies) wood hemicelluloses have been obtained by the noncatalytic and catalytic oxidative delignification in the acetic acid-water-hydrogen peroxide medium in a processing time of 3-4 h and temperatures of 90-100 °C. In the catalytic process, the H2SO4, MnSO4, TiO2, and (NH4)6Mo7O24 catalysts have been used. A polysaccharide yield of up to 11.7 wt% has been found. The hemicellulose composition and structure have been studied by a complex of physicochemical methods, including gas and gel permeation chromatography, Fourier-transform infrared spectroscopy, and thermogravimetric analysis. The galactose:mannose:glucose:arabinose:xylose monomeric units in a ratio of 5:3:2:1:1 have been identified in the hemicelluloses by gas chromatography. Using gel permeation chromatography, the weight average molar mass Mw of hemicelluloses has been found to attain 47,654 g/mol in noncatalytic delignification and up to 42,793 g/mol in catalytic delignification. Based on the same technique, a method for determining the α and k parameters of the Mark-Kuhn-Houwink equation for hemicelluloses has been developed; it has been established that these parameters change between 0.33-1.01 and 1.57-472.17, respectively, depending on the catalyst concentration and process temperature and time. Moreover, the FTIR spectra of the hemicellulose samples contain all the bands characteristic of heteropolysaccharides, specifically, 1069 cm-1 (C-O-C and C-O-H), 1738 cm-1 (ester C=O), 1375 cm-1 (-C-CH3), 1243 cm-1 (-C-O-), etc. It has been determined by the thermogravimetric analysis that the hemicelluloses isolated from spruce wood are resistant to heating to temperatures of up to ~100 °C and, upon further heating, start destructing at an increasing rate. The antioxidant activity of the hemicelluloses has been examined using the compounds simulating the 2,2-diphenyl-2-picrylhydrazyl free radicals.

Improving the nutritional value and digestibility of wheat straw, rice straw, and corn cob through solid state fermentation using different Pleurotus species.

BACKGROUND: The growing food-feed-fuel competition, declining availability of traditional feeds, higher prices, and the urgent need to provide long-term sustainability for animal production have all triggered global research into the optimum extraction of energy and nutrients from lignin-rich plant biomass. Recent studies have shown that the Pleurotus species of white rot fungus can selectively degrade lignin in lignin-rich plant biomass; however, its effectiveness in selectively degrading lignin depends on the type of substrate and species of fungus. This study was therefore designed to treat wheat straw, rice straw, and corn cob, with Pleurotus eryngii, P. ostreatus, and P. florida for 30 days under solid-state fermentation, to identify a promising fungus-substrate combination for the selective degradation of lignin and optimal improvement in the nutritional value and digestibility of each substrate. RESULTS: The type of fungus strongly influenced (P < 0.01) selectivity in lignin degradation, and the level of improvement in crude protein (CP), in vitro dry matter digestibility (IVDMD), and in vitro gas production (IVGP), in wheat straw, rice straw, and corn cob. Fungus-substrate interaction data revealed that P. ostreatus caused maximum (P < 0.05) degradation of lignin, and greater (P < 0.05) improvement in CP, IVDMD, and IVGP in wheat straw and rice straw. The lowest (P < 0.05) degradation of lignin and improvement in CP, IVDMD, and IVGP was caused by P. eryngii in corn cob. Among the fungi, the maximum (P < 0.05) degradation of lignin, and greater (P < 0.05) improvement in CP, IVDMD, and IVGP were caused by P. florida as compared with those of P. ostreatus and P. eryngii. CONCLUSION: The results highlight significant influence of fungus-substrate combination for selective lignin degradability and the consequent improvement in the nutritional value of the substrates. Maximum selective lignin degradability and improvement in nutritional value and digestibility was caused by P. ostreatus in wheat straw and in rice straw, and by P. florida in corn cob. © 2021 Society of Chemical Industry.

Comparison of performances of different fungal laccases in delignification and detoxification of alkali-pretreated corncob for bioethanol production.

The performance of the alkaline fungal laccase PIE5 (pH 8.5) in the delignification and detoxification of alkali-pretreated corncob to produce bioethanol was evaluated and compared with that of the neutral counterpart (rLcc9, 6.5), with the acidic laccase rLacA (4.0) was used as an independent control. Treatment with the three laccases facilitated bioethanol production compared with their respective controls. The lignin contents of alkali-pretreated corncob reduced from 4.06%, 5.06%, and 7.80% to 3.44%, 3.95%, and 5.03%, after PIE5, rLcc9, and rLacA treatment, respectively. However, the performances of the laccases were in the order rLacA > rLcc9 > PIE5 in terms of decreasing total phenol concentration (0.18, 0.36, and 0.67 g/l), boosting ethanol concentration (8.02, 7.51, and 7.31 g/l), and volumetric ethanol productivity (1.34, 0.94, and 0.91 g/l hr), and shortening overall fermentation time. Our results would inform future attempts to improve laccases for ethanol production. Furthermore, based on our data and the fact that additional procedures, such as pH adjustment, are needed during neutral/alkaline fungal laccase treatment, we suggest acidic fungal laccases may be a better choice than neutral/alkaline fungal laccases in bioethanol production.

Enhancement of lignocellulosic biomass anaerobic digestion by optimized mild alkaline hydrogen peroxide pretreatment for biorefinery applications.

Lignocellulosic energy crops are promising feedstocks for producing renewable fuels, such as methane, that can replace diminishing fossil fuels. However, there is a major handicap in using lignocellulosic sources to produce biofuels, which is their low biodegradability. In this study, the application and the optimization of a lignocellulose pretreatment process, named alkaline hydrogen peroxide, was investigated for the enhancement of methane production from the energy crop switchgrass. Four independent process variables, solid content (3-7%), reaction temperature (50-100 °C), H2O2 concentration (1-3%), and reaction time (6-24 h), and three response variables, soluble reducing sugar, soluble chemical oxygen demand, and biochemical methane potential were used in process optimization and modeling. The optimization was performed by two different approaches as maximum methane production and cost minimization. The optimum conditions for the highest methane production were found as 6.65 wt% solid content, 50.6 °C reaction temperature, 2.94 wt% H2O2 concentration, and 16.05 h reaction time. The conditions providing the lowest cost were 6.43 wt% solid content, 50 °C reaction temperature, 1.83 wt% H2O2 concentration, and 6.78 h reaction time. For maximum methane production and cost minimization, specific methane yields of 338.52 mL CH4/g VS and 291.34 mL CH4/g VS were predicted with 62.4 % and 39.8 % enhancements compared to untreated switchgrass, respectively. Finally, it was found that the predicted methane production for the maximum methane production represents 77 % of the theoretical methane yield and 82.22 % energy recovery.

Green Deep Eutectic Solvents for Microwave-Assisted Biomass Delignification and Valorisation.

Aiming to fulfil the sustainability criteria of future biorefineries, a novel biomass pretreatment combining natural deep eutectic solvents (NaDESs) and microwave (MW) technology was developed. Results showed that NaDESs have a high potential as green solvents for lignin fractionation/recovery and sugar release in the following enzymatic hydrolysis. A new class of lignin derived NaDESs (LigDESs) was also investigated, showing promising effects in wheat straw delignification. MW irradiation enabled a fast pretreatment under mild condition (120 °C, 30 min). To better understand the interaction of MW with these green solvents, the dielectric properties of NaDESs were investigated. Furthermore, a NaDES using the lignin recovered from biomass pretreatment as hydrogen bond donor was prepared, thus paving the way for a "closed-loop" biorefinery process.

Atomic force microscopy imaging of delignified secondary cell walls in liquid conditions facilitates interpretation of wood ultrastructure.

Deep understanding of the physicochemical and structural characteristics of wood at the nanoscale is essential for improving wood usage in biorefining and advancing new high performance materials design. Herein, we use in situ atomic force microscopy and a simple delignification treatment to elucidate the nanoscale architecture of individual secondary cell wall layers. Advantages of this approach are: (i) minimal sample preparation that reduces the introduction of potential artifacts; (ii) prevention of structural rearrangements due to dehydration; (iii) increased accessibility to structural details masked by the lignin matrix; and (iv) possibility to complement results with other analytical techniques without sample manipulation. The methodology permits the visualization of parallel and helicoidally arranged microfibril aggregates in the S1 layer and the determination of lignin contribution to microfibril aggregates forming S2 layers. Cellulose and hemicelluloses constitute the core of the aggregates with a mean diameter of approximately 19 nm, and lignin encloses the core forming single structural entities of about 30 nm diameter. Furthermore, we highlight the implications of sample preparation and imaging parameters on the characterization of microfibril aggregates by AFM.

Bioethanol Production from Vineyard Waste by Autohydrolysis Pretreatment and Chlorite Delignification via Simultaneous Saccharification and Fermentation.

In this paper, the production of a second-generation bioethanol from lignocellulosic vineyard cutting wastes was investigated in order to define the optimal operating conditions of the autohydrolysis pretreatment, chlorite delignification and simultaneous saccharification and fermentation (SSF). The autohydrolysis of vine-shoot wastes resulted in liquors containing mainly a mixture of monosaccharides, degradation products and spent solids (rich in cellulose and lignin), with potential utility in obtaining valuable chemicals and bioethanol. The autohydrolysis of the vine-shoot wastes was carried out at 165 and 180 °C for 10 min residence time, and the resulted solid and liquid phases composition were analysed. The resulted liquid fraction contained hemicellulosic sugars as a mixture of alpha (α) and beta (ß) sugar anomers, and secondary by-products. The solid fraction was delignified using the sodium chlorite method for the separation of lignin and easier access of enzymes to the cellulosic sugars, and then, converted to ethanol by the SSF process. The maximum bioethanol production (6%) was obtained by autohydrolysis (165 °C), chlorite delignification and SSF process at 37 °C, 10% solid loading, 72 h. The principal component analysis was used to identify the main parameters that influence the chemical compositions of vine-shoot waste for different varieties.

Improving enzymatic hydrolysis efficiency of corncob residue through sodium sulfite pretreatment.

The effects of sodium sulfite pretreatment on the delignification rate, cellulose content, enzymatic hydrolysis efficiency, and glucose yield of corncob residues (CCR) were investigated. The optimum pretreatment conditions were as follows: 12% sodium sulfite, with a pH value of 7, a temperature of 160 °C, and a holding time of 20 min. Under the optimal conditions, the cellulose content in the pretreated residue was 85.17%, and sodium lignosulfonate with a sulfonation degree of 0.677 mmol/g was obtained in the waste liquids. A delignification rate of 77.45% was also achieved after the pretreatment. Enzymatic hydrolysis of pretreated CCR was carried out with cellulase (5 FPU/g substrate) and ß-glucosidase (10 IU/g substrate) for 48 h. The untreated CCR were hydrolyzed using cellulase (20 FPU/g substrate) and ß-glucosidase (10 IU/g substrate) for 48 h. The comparison results showed that sodium sulfite pretreatment improved the enzymatic hydrolysis efficiency and glucose yield, which increased by 28.80% and 20.10%, respectively. These results indicated that despite the application of low cellulase dosage, high enzymatic hydrolysis efficiency substrate could be produced, and the sodium lignosulfonate which can be used for oilfields and concrete additives was obtained from the sodium sulfite-pretreated CCR.

Bacterial laccases: promising biological green tools for industrial applications.

Multicopper oxidases (MCOs) are a pervasive family of enzymes that oxidize a wide range of phenolic and nonphenolic aromatic substrates, concomitantly with the reduction of dioxygen to water. MCOs are usually divided into two functional classes: metalloxidases and laccases. Given their broad substrate specificity and eco-friendliness (molecular oxygen from air as is used as the final electron acceptor and they only release water as byproduct), laccases are regarded as promising biological green tools for an array of applications. Among these laccases, those of bacterial origin have attracted research attention because of their notable advantages, including broad substrate spectrum, wide pH range, high thermostability, and tolerance to alkaline environments. This review aims to summarize the significant research efforts on the properties, mechanisms and structures, laccase-mediator systems, genetic engineering, immobilization, and biotechnological applications of the bacteria-source laccases and laccase-like enzymes, which principally include Bacillus laccases, actinomycetic laccases and some other species of bacterial laccases. In addition, these enzymes may offer tremendous potential for environmental and industrial applications.

Enhanced production, one-step affinity purification, and characterization of laccase from solid-state culture of Lentinus tigrinus and delignification of pistachio shell by free and immobilized enzyme.

Laccase mediated bio-delignification has shown promising results for the removal of lignin from bio-wastes and for providing a sustainable future for using of lignocellulosic materials in different industries. This study reports an extracellular laccase from Lentinus tigrinus with delignification capability. The production of laccase was enhanced through a solid-state fermentation on the pistachio shell bio-waste to 172.0 U mg-1 (8.2-fold) by one-factor-at-a-time optimizing of fermentation conditions. Laccase was purified using a new synthetic affinity resin yielding a specific activity of 543.6 U mg-1 and a 23.9-fold purification. The purified laccase was then immobilized covalently on the large pore magnetic SBA-15. Compared to free enzyme, immobilized enzyme maintained more stable at pH 2.0-11.0 and 25-55 °C, and against organic solvents, surfactants, metal ions, and inhibitors. The activity of both forms of the enzyme was increased with Cu2+, Ca+2, cetyltrimethylammonium bromide, and ethyl acetate. A 0.72 V redox potential caused enzyme specificity to various substrates. 80% of lignin content of the bio-waste was removed by 50 U mL-1 of immobilized enzyme after 8 h fermentation and delignification efficiency was greatly increased by applying higher enzyme dosages, surfactants, and organic solvents. In addition, residual activity was more than 50% after 20 cycles of delignification. The results of delignification were confirmed by GC-MS, SEM, and composition analysis of pistachio shells. This study illustrated the notable promise of the enzyme for biotechnological and environmental applications.

Glucuronoyl esterases: diversity, properties and biotechnological potential. A review.

Glucuronoyl esterases (GEs) belonging to the carbohydrate esterase family 15 (CE15) are involved in microbial degradation of lignocellulosic plant materials. GEs are capable of degrading complex polymers of lignin and hemicellulose cleaving ester bonds between glucuronic acid residues in xylan and lignin alcohols. GEs promote separation of lignin, hemicellulose and cellulose which is crucial for efficient utilization of biomass as an energy source and feedstock for further processing into products or chemicals. Genes encoding GEs are found in both fungi and bacteria, but, so far, bacterial GEs are essentially unexplored, and despite being discovered >10 years ago, only a limited number of GEs have been characterized. The first laboratory scale example of improved xylose and glucuronic acid release by the synergistic action of GE with cellulolytic enzymes was only reported recently (improved C5 sugar and glucuronic acid yields) and, until now, not much is known about their biotechnology potential. In this review, we discuss the diversity, structure and properties of microbial GEs and consider the status of their action on natural substrates and in biological systems in relation to their future industrial use.

Lignocellulosic biomass to biofuels and biochemicals: A comprehensive review with a focus on ethanol organosolv pretreatment technology.

Lignocellulosic biomass is one of the potential feedstocks to produce second-generation cellulosic ethanol and biochemicals. To enhance the enzymatic digestibility of lignocellulosic biomass for efficient enzymatic saccharification, a variety of pretreatment methods have been studied. Among these, organosolv pretreatment using ethanol is a promising pretreatment method owing to its inherent advantages, such as low solvent cost, lack of toxicity, the ability to retain most cellulose fraction in substrates for enzymatic hydrolysis, coproduction of high-purity lignin and hemicellulosic sugars, easy solvent recovery, and reuse. In this review, the research progress regarding different types of ethanol organosolv pretreatment processes has been summarized in terms of methods, substrate properties, reaction mechanisms, delignification kinetic as well as the impact of pretreatment methods on the enzymatic digestibility. Attempts are also made to provide insights into the complete utilization of lignocellulosic biomass to achieve high potential revenues. Though some ethanol organosolv processes have been studied or are being developed towards commercialization, ethanol organosolv pretreatment is still facing some challenges. Finally, the direction for future work is given to develop a proper ethanol organosolv pretreatment for commercialization.