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The present study reports the combined enzymatic production efficiency of thermophilic fungus Thermomyces lanuginosus VAPS25 using a combinatory artificial intelligence-based tool, resulting in 2.7 IU/ml, 5.2 IU/ml, and 18.85 U/ml activity of endoglucanase, amylase, and lipase, respectively with good thermostability at 90 °C (pH 8-10). Interestingly, the metal ions viz. Cu 2+ and Mg 2+ increased the endoglucanase activity to 5 folds, i.e.,5.6 IU/ml compared to control. Further, the amylase and lipase activity was also enhanced by Fe 2+ and Co 2+ to 5.4 IU/ml and 19.57 U/ml, respectively. Additionally, the deinking efficiency was improved by 68.9%, 42.7%, and 52.8% by endoglucanase, amylase, and lipase, respectively, while the consortium increased the deinking efficiency to 72.7%. The bio-bleached paper strength parameters such as burst index, breaking length, tear index, and tensile index of sheets were significantly improved by 1.38%, 13.54%, 7.54%, and 20.88%, respectively. These enzymes at an industrial scale would help develop an economical paper recycling process., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

In this work, effects of sulfomethylated lignins (SLs) prepared from masson pine (SL M ) and poplar (SL P ) on enzymatic hydrolysis and cellulase-lignin interaction were comparatively investigated. The results showed that both SL M and SL P significantly promoted the substrate enzymatic digestibility. The total sugar yield increased from 38.6% to 74.4% and ∼ 100%, respectively at 10 FPU/g-cellulose of cellulase dosage. The protein content in hydrolysate linearly increased with the addition of SL (0 - 1.6 g/g-substrate lignin), which suggested the competitive adsorption of cellulase may occur to substrate lignin and SLs. Further structural analysis of lignin revealed the high S/(V + H) ratio was directly related to the high enzymatic saccharification efficiency. The strong interaction between SL and cellulase decreased the nonproductive adsorption of cellulase onto substrate lignin and increased the accessibility of cellulase to carbohydrate, which was considered to be the key factor for the improvement of substrate enzymatic digestibility., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

An integration of different pretreatments is important to overcome recalcitrance and realize efficient bioconversion of lignocellulosic biomass. This study aims at the effects of combination of hydrothermal pretreatment and different chemi-mechanical pretreatments on enzymatic hydrolysis, and understanding the enzymes adsorption mechanism. The combination of hydrothermal and chemi-mechanical pretreatments effectively improved the enzymatic hydrolysis of poplar substrates, in which the enzymatic hydrolysis of substrates pretreated by hydrothermal pretreatment + Fenton pretreatment + mechanical refining (HFM) was the highest (92.39% of glucose conversion yield, and 20.88 g/L of glucose concentration). The substrates' main characteristics were obviously changed after combined pretreatments, such as swelling ability and specific surface area of substrates were increased. The Langmuir adsorption model (R 2  > 0.98) and pseudo second-order adsorption kinetic model (R 2 ≈1) were well suitable to describe the adsorption of enzymes on substrates, meanwhile the adsorption mechanism was summarized., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The aim of this work was to develop a feasible ball mill-assisted alkaline peroxide pretreatment followed by stepwise hydrolysis to improve the yield of xylo-oligosaccharides (XOS) and fermentable sugars. The hydrogen peroxide charge, ball-milling time, and solid-to-liquid ratio affected the compositions, particle sizes, morphology, and crystallinity of the corn stover, directly improving the following hydrolytic efficiency. The optimal pretreatment was with 0.45 g/g (H 2 O 2 : substrate) and 1:3 solid-to-liquid ratio (w/v) for 1.0 h ball-milling, resulting in 84.29% delignification. Physicochemical properties of the pretreated samples were characterized and their correlations to the enzymatic hydrolysis were revealed. Compared with one-step cellulase hydrolysis, the two-step xylanase-cellulase hydrolysis of the pretreated corn stover showed significant advance in preparing XOS, producing 69.65% (on the base of xylan content in pretreated sample) of XOS, along with 20.55% xylose, 68.94% glucose, and 21.15% gluco-oligosaccharides. The yield of XOS was 2-7 times higher than those in previous studies., (Copyright © 2021. Published by Elsevier Ltd.)

The high content of lignin in bamboo is considered as the major obstacle for its biorefining. In this work, a green, lignin-selective, and recyclable solvent of phenoxyethanol was coupled with acid solution to deconstruct recalcitrant structure of bamboo residues (BR) to boost its enzymatic digestibility. Results showed phenoxyethanol has excellent lignin-removal ability from 29.4% to 91.6% when phenoxyethanol:acid ratios increased from 0:1 to 4:1 at 120 °C. 82.5%-87.8% of cellulose was preserved in pretreated BR. The enzymatic digestibility of BR significantly improved from 20.0% to 91.3% when it was pretreated under optimized conditions. With lower enzyme dosages (10 FPU/g) and 5 recycled using of pretreatment liquor, pretreated BR still showed a good enzymatic digestibility of 67.4%-93.7% and 67.1-76.8%, respectively. Examination of physicochemical changes revealed that improvements to accessibility, reduction of crystallite size, decrease of surface lignin and hydrophobicity for pretreated BR showed positive correlations (R 2  > 0.7) with their enzymatic digestibility., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Viscosity control and reactivity enhancement are of practical importance for high-quality dissolving pulp manufacturing. In this work, we demonstrate a two-step activating process consisting of a phosphotungstic acid (PTA)-assisted prerefining (PTA/R pretreatment), followed by cellulase treatment for this purpose. The cellulase adsorption can increase from 29.1% to 49.7% as a result of PTA/R pretreatment (8000 r at 90 °C). The viscosity of the resultant pulp decreases from 665 to 430 mL/g, while its Fock reactivity increases from 31.5% to 74.4% under a low-loading cellulase treatment (0.5 mg cellulase /g odp), which mainly due to the fact that the PTA/R pretreatment can increase fiber accessibility and viscosity control, thus facilitating cellulase adsorption and reaction efficiency. Moreover, PTA also shows a high recyclability/ reusability (more than 86%) during the PTA/R pretreatment. Therefore, the new proposed two-step activating process provides a green, and efficient pathway for large-scale manufacturing of high-quality dissolving pulp., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Enzymatic hydrolysis of three pre-treated lignocellulosic biomasses -LCB- (wheat straw-WS-, corn stover-CSV- and cardoon stems -CS-) is studied. These biomasses were pre-treated by two methods: diluted sulfuric acid and acid ethanol-water extraction at six severity levels (H values). Pretreated solid fractions were hydrolyzed with commercial enzyme cocktails at standard conditions. A first-order kinetic fractal model was fitted to the experimental results. This model accurately describes the hydrolysis of all biomasses at all pre-treatment conditions studied. The results show that the formal first-order kinetic constant k depends on the biomass nature. The hydrolysis rate increases as the pre-treatment severity does, while the fractal exponent value h decreases. With these pre-treatments, and in terms of k and h, WS is highly reactive and, at medium H with EW pretreatment, highly accessible; CSV has a low reactivity and high accessibility and CS has the lowest reactivity and an increasing accessibility as severity rises., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Understanding the adsorption behavior and the interaction between substrates and enzymes are critical to improving enzymatic hydrolysis efficiency and reducing bioconversion cost. Herein, the adsorption of cellulase on wet ground corncob residues was studied, and the effects of main characteristics of wet ground corncob residues on adsorption capacity were quantitatively analyzed with the combination of principal component analysis and multiple linear regression models. The results showed that the adsorption of cellulase on wet ground corncob residues was fitted well with Langmuir isotherm adsorption and pseudo second-order kinetics model, the adsorption rate and adsorption capacity were greatly enhanced with increasing grinding time; the multiple linear regression models describing the relationship between main characteristics of corncob residues and adsorption capacity to cellulase were established; the significance of these characteristics were in the following order: average particle size, crystallinity index, specific surface area, surface lignin concentration, water retention value, and surface charge density., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Most additives that capable of enhancing enzymatic hydrolysis of lignocellulose are petroleum-based, which are not easy to recycle with poor biodegradability. In this work, highly recyclable and biodegradable sodium caseinate (SC) was used to enhance lignocellulosic hydrolysis with improved cellulase recyclability. When the pH decreased from 5.5 to 4.8, more than 96% SC could be precipitated from the solution and recovered. Adding SC increased enzymatic digestibility of dilute acid pretreated eucalyptus (Eu-DA) from 39.5% to 78.2% under Eu-DA loading of 10 wt% and pH = 5.5, and increase cellulase content in 72 h hydrolysate from only 15.2% of the original to 60.0%, which facilitated the recovery of cellulases through re-adsorption by fresh substrates. With multiple cycles of re-adsorption, application of SC not only increased the sugar yield of Eu-DA by 95.5%, but also reduced cellulase loading by 40%., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Enzymatic hydrolysis of lignocellulose to produce bioethanol by cellulase is an important method to alleviate the energy crisis. In this paper, in order to overcome the shortcomings of low efficiency, high cost and easy deactivation of cellulase in the process of bio-refinery, pH-responsive lignin-based magnetic nanoparticles (Fe 3 O 4 /LSQA) were synthesized to immobilize and recover cellulase. It was shown that a high immobilization ratio of 55.52% for cellulase was obtained. Meanwhile, the desorption ratio was 68.27% by adjusting the pH of the system. After five reusing cycles, the desorbed cellulase retained 31.79% of the relative activity due to the pH responsiveness of Fe 3 O 4 /LSQA. These results not only provide a new idea for the recycling of cellulase, but also broaden the application of industrial lignin and increase the extra value., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The isoelectric point (pI) of lignin-based surfactant is an important factor in the enhancement on the enzymatic hydrolysis of lignocellulose. In this work, lignin carboxylate (LC) and quaternary ammonium lignin carboxylates (LCQ-x, x%: the mass ratio of quaternizing agent to enzymatic hydrolysis lignin) with different isoelectric points were synthesized. LC or LCQ-x with pI significantly lower or higher than 4.8 reduced the non-productive adsorption of cellulase on lignin, but for the significant inhibitory effect on cellulase activity, their enhancements on the enzymatic hydrolysis of lignocellulose were not remarkable. However, LCQ-x with pI around 4.8 preserved the cellulase activity, and significantly reduced the non-productive adsorption of cellulase, therefore remarkably enhanced the enzymatic hydrolysis. 2 g/L LC, LCQ-40 (pI = 5.0) and LCQ-100 (pI = 9.2) increased the enzymatic digestibility of pretreated eucalyptus from 35.2% to 53.4%, 95.3% and 60.4% respectively. In addition, for the excellent pH-response performance, LCQ could be efficiently recovered after enzymatic saccharification., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Enzymatic pretreatment has shown great potential in making the disintegration of cellulosic fibers to fibrils cost-effectively and environmental-friendly. In this study, an extensive commercial endoglucanase was used to pretreat cellulosic fibers for fibrillation. The pretreatment time and the enzyme dosage were optimized using response surface methodology. A 100% fiber recovery was obtained at endoglucanase usage of 9.0 mg/g (substrate) and pretreatment time of less than 3.0 h. A highly fibrillated and fractured surface of pretreated fibers was observed after 0.5 h of pretreatment compared to native fibers. Meanwhile, the progressive deconstruction of cellulosic fibers was occurred with the enzymatic pretreatment time increasing. The degree of deconstruction of fibers was evidenced by changes of the fiber microstructure, such as the inter-/intra-molecular H-bonds, the β-1,4-glucosidic linkages, crystallinity and crystallite size. These discoveries provide new insights into a more efficient and economic pretreatment methods for the disintegration of fibrils from cellulosic fibers., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Dissolving pulp is an important source of cellulose raw material, and its key quality parameter is the Fock reactivity for viscose rayon application. Cellulase treatment is an effective method for improving the Fock reactivity of kraft-based dissolving pulp. In this study, a novel process concept of improving the cellulase treatment for this purpose was developed, and it consists of mechanical pre-refining and PDADMAC-assisted cellulase treatment. The hypothesis is based on: 1) opening up the fiber structures to improve the cellulase accessibility by pulp prerefining, 2) the addition of cationic poly DADMAC to the subsequent cellulase stage enhances the cellulase adsorption onto anionic fibers due to favorable electrostatic interactions. The results showed that the Fock reactivity of the resultant pulp from the combined treatment is much higher than that of the control, yet, achieved at a much lower cellulase dosage., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The reactivity enhancement and viscosity control are of practical importance during the manufacture of high-quality cellulose (also known as dissolving pulp). In the study, the concept of using phosphotungstic acid (HPW) for this purpose was demonstrated. The Fock reactivity of resultant pulp increased from 49.1% to 74.1% after the HPW catalytic treatment at a dosage of 86.4 mg HPW/g odp. The improved results can be attributed to the increased fiber accessibility, thanks to the favorable fiber morphologic changes, such as increased pore volume/size, water retention value and specific surface area. HPW can be readily recycled/reused by evaporating method, where maintaining 87.1% catalytic activity after six recycle times. The HPW catalytic treatment concept may provide a green alternative for the manufacture of high-quality dissolving pulp., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The effective separations of cellulose and hemicelluloses from cellulosic fibers are the prerequisite for creating high-value to the abundant and green cellulose materials. In this study, the process concept of cellulase pretreatment, followed by a cold caustic extraction (CCE) was investigated for a softwood sulfite pulp. The results showed that the cellulase pretreatment led to favorable fiber morphological changes, including the increases of the specific surface area (SSA), pore volume and diameter, and the water retention value (WVR). These changes can induce more pronounced fiber swelling in the subsequent CCE process so that the hemicelluloses removal is enhanced. After the cellulase pretreatment (cellulase dosage of 1 mg/g) and CCE process, the cellulose purity was as high as 97.49%, while the hemicelluloses removal selectivity reached 76.42%., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Many positive effects of additives on enzymatic hydrolysis of lignocellulosic materials have been investigated, but limited information has been reported on the use of long-chain fatty alcohols (LFAs) for enzymatic hydrolysis by biospired models. In this study, effects of LFAs on enzymatic hydrolysis were evaluated using biomimetic film asa structure model. LFAs clearly improved the digestibility of cellulose film from 65.1% to 77.9%, which was higher than that the digestibility of lignin-cellulose film from 53.9% to 66.2%. Further study indicated that the promotion ascribed to the effect of LFAs, which might provide more active points of chemical reaction and keep the stability of cellulase. Moreover, the digestibility of lignin-cellulose film increased by 12.3%, which might because the denaturation and nonproductive adsorption of cellulase were well prevented by supplementation of LFAs. An efficient strategy was developed to improve the enzymatic hydrolysis efficiency in the study of lignocellulosic bioconversion., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Effects of nonionic surfactants on enzymatic hydrolysis of Avicel at different agitation rates and solid loadings and the mechanism were studied. Nonionic surfactants couldn't improve the enzymatic hydrolysis efficiency at 0 and 100rpm but could enhance the enzymatic hydrolysis significantly at high agitation rate (200 and 250rpm). Cellulase was easily deactivated at high agitation rate and the addition of nonionic surfactants can protect against the shear-induced deactivation, especially when the cellulase concentration was low. When 25mg protein/L of cellulase solution was incubated at 200rpm for 72h, the enzyme activity increased from 36.0% to 89.5% by adding PEG4600. Moreover nonionic surfactants can compete with enzyme in air-liquid interface and reduce the amount of enzyme exposed in the air-liquid interface. The mechanism was proposed that nonionic surfactants could enhance the enzymatic hydrolysis of Avicel by reducing the cellulase deactivation caused by shear force and air-liquid interface., (Copyright © 2017. Published by Elsevier Ltd.)

Some zwitterionic surfactants exhibit upper critical solution temperature (UCST) in aqueous solutions. For the zwitterionic surfactant solution mixed with cellulase, when its temperature is below UCST, the cellulase can be recovered by coprecipitation with zwitterionic surfactant. In this work, 3-(Hexadecyldimethylammonio) propanesulfonate (SB3-16) was selected to enhance the enzymatic hydrolysis of lignocelluloses and recover the cellulase. After adding 2mmol/L of SB3-16, the enzymatic digestibility of eucalyptus pretreated by dilute acid (Eu-DA) and by sulfite (Eu-SPORL) increased from 27.9% and 35.1% to 72.6% and 89.7%, respectively. The results showed that SB3-16 could reduce the non-productive adsorption of cellulase on hydrophobic interface, while it did not significantly inhibit the activity of cellulase. For the solution contained 1wt% SB3-16 and 200mg protein/L CTec2 cellulase, 55.2% of protein could be recovered by cooling. The filter paper activity of the recovered cellulase was 1.93FPU/mg protein, which was 95.8% of its initial activity., (Copyright © 2017. Published by Elsevier Ltd.)

Dilute acid pre-extraction enhanced the mechanically refined poplar pulp substrates' enzymatic hydrolysis efficiency obviously. The results showed that the surface lignin distribution was changed significantly in residual wood chips and pulp substrates, and the surface lignin distribution showed important impact on the following enzymatic hydrolysis. Acid pre-extraction can lead to a redistribution of lignin in fiber cell walls, i.e., the lignin was degraded and migrated to fiber surface in the form of re-deposited lignin and pseudo-lignin. However, higher pre-extraction intensity was not desired due to the formation of redeposited lignin and pseudo-lignin. This study will help to reach a deeper understanding on the lignin distribution in the view of molecular and ultrastructure, and promote the development of a cost-efficient pretreatment strategy for biomass processing., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The aim of this study was to investigate a combined process involving pulp fractionation and cellulase treatment of each fraction for improving the molecular weight distribution (MWD) and reactivity of a kraft-based dissolving pulp. Three pulp fractions, namely long-fiber, mid-fiber and short-fiber fractions (LF, MF and SF, respectively), were used as the substrates. The results showed that the SF had the highest accessibility, lowest viscosity, and highest cellulase adsorption capacity, while the opposite was true for the LF. At a given viscosity, the combined process led to a lower polydispersity index (3.71 vs 4.98) and a higher Fock reactivity (85.6% vs 76.3%), in comparison to the conventional single-stage cellulase treatment., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Cost-effectiveness is vital for enzymatic treatment of dissolving pulp towards industrial application. The strategy of cellulase recycling with fresh cellulase addition was demonstrated in this work to activate the dissolving pulp, i.e. decreasing viscosity and increasing Fock reactivity. Results showed that 48.8-35.1% of cellulase activity can be recovered from the filtered liquor in five recycle rounds, which can be reused for enzymatic treatment of dissolving pulp. As a result, the recycling cellulase with addition fresh cellulase of 1mg/g led to the pulp of viscosity 470mL/g and Fock reactivity 80%, which is comparable with cellulase charge of 2mg/g. Other pulp properties such as alpha-cellulose, alkaline solubility and molecular weight distribution were also determined. Additionally, a zero-release of recycling cellulase treatment was proposed to integrate into the dissolving pulp production process., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

A modified process consisting of an initial mechanical refining (R) followed by a low-alkali (5.5% NaOH) cold caustic extraction (CCE) and finally an endoglucanase (EG) treatment (R-5.5%CCE-EG) was investigated for upgrading paper-grade pulp to dissolving pulp. Results showed that compared to the conventional process (9%CCE-EG), the modified process can decrease the alkali concentration (from 9% to 5.5%) to achieve a similar hemicelluloses removal while simultaneously enhancing the Fock reactivity (from 62.2% to 81.0%). The improved results were due to the fact that the mechanical refining resulted in favorable fiber morphological changes, including increased pore volume/size, water retention value and specific surface area. Consequently, the hemicelluloses removal was enhanced even under the subsequent low-alkali CCE condition. A synergic effect of refining, low alkali concentration and enzymatic activation was responsible for the higher reactivity of resulting dissolving pulp., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Water-soluble lignin-based polyoxyethylene ether (EHL-PEG), prepared from enzymatic hydrolysis lignin (EHL) and polyethylene glycol (PEG1000), was used to improve enzymatic hydrolysis efficiency of corn stover. The glucose yield of corn stover at 72h was increased from 16.7% to 70.1% by EHL-PEG, while increase in yield with PEG4600 alone was 52.3%. With the increase of lignin content, EHL-PEG improved enzymatic hydrolysis of microcrystalline cellulose more obvious than PEG4600. EHL-PEG could reduce at least 88% of the adsorption of cellulase on the lignin film measured by quartz crystal microbalance with dissipation monitoring (QCM-D), while reduction with PEG4600 was 43%. Cellulase aggregated at 1220nm in acetate buffer analyzed by dynamic light scattering. EHL-PEG dispersed cellulase aggregates and formed smaller aggregates with cellulase, thereby, reduced significantly nonproductive adsorption of cellulase on lignin and enhanced enzymatic hydrolysis of lignocelluloses., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Cellulase treatment for decreasing viscosity and increasing Fock reactivity of dissolving pulp is a promising approach to reduce the use of toxic chemicals, such as hypochlorite in the dissolving pulp manufacturing process in the industry. Improving the cellulase treatment efficiency during the process is of practical interest. In the present study, the concept of using cationic polyacrylamide (CPAM) to enhance the cellulase treatment efficiency was demonstrated. This was mainly attributed to the increased cellulase adsorption onto cellulose fibers based on the patching/bridging mechanism. Results showed that the cellulase adsorption was increased by about 20% with the addition of 250 ppm of CPAM under the same conditions as those of the control. It was found that the viscosity decrease and Fock reactivity increase for the cellulase treatment was enhanced from using CPAM. The CPAM-assisted cellulase treatment concept may provide a practical alternative to the present hypochlorite-based technology for viscosity control in the industry., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The presence of lignin in lignocellulosic biomass is correlated with its enzymatic digestibility. Their correlation and mechanism have been investigated widely but have not been elucidated clearly. In this study, hydrophilic sulfonated lignin and hydrophobic kraft lignin were introduced into the enzymatic hydrolysis process to investigate their effects on the enzymatic digestibility of different pretreated lignocellulose. The influence of lignin addition on the enzymatic digestibility varied with both introduced lignin type and the pretreatment methods of substrates. Slight enhancement of enzymatic hydrolysis was observed for all substrates by adding kraft lignin. The addition of sulfonated lignin could effectively improve the enzymatic digestibility of green liquor and acidic bisulfite pretreated materials, but had little effect on sulfite-formaldehyde pretreated samples. The enzymatic digestibility of green liquor pretreated masson pine increased from 42% without lignin addition to 75% with 0.3g/g-substrate sulfonated lignin addition., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

For enzymatic treatment of dissolving pulp, there is a need to improve the process to facilitate its commercialization. For this purpose, the high consistency cellulase treatment was conducted based on the hypothesis that a high cellulose concentration would favor the interactions of cellulase and cellulose, thus improves the cellulase efficiency while decreasing the water usage. The results showed that compared with a low consistency of 3%, the high consistency of 20% led to 24% increases of cellulase adsorption ratio. As a result, the viscosity decrease and Fock reactivity increase at consistency of 20% were enhanced from 510 mL/g and 70.3% to 471 mL/g and 77.6%, respectively, compared with low consistency of 3% at 24h. The results on other properties such as alpha cellulose, alkali solubility and molecular weight distribution also supported the conclusion that a high consistency of cellulase treatment was more effective than a low pulp consistency process., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Three strategies were presented to achieve high solids loading while maximizing carbohydrate conversion, which are fed-batch, splitting/thickening, and clarifier processes. Enzymatic hydrolysis was performed at water insoluble solids (WIS) of 15% using washed dilute-acid pretreated corn stover. The carbohydrate concentration increased from 31.8 to 99.3g/L when the insoluble solids content increased from 5% to 15% WIS, while the final carbohydrate conversion was decreased from 78.4% to 73.2%. For the fed-batch process, a carbohydrate conversion efficiency of 76.8% was achieved when solid was split into 60:20:20 ratio, with all enzymes added first. For the splitting/thickening process, a carbohydrate conversion of 76.5% was realized when the filtrate was recycled to simulate a steady-state process. Lastly, the clarifier process was evaluated and the highest carbohydrate conversion of 81.4% was achieved. All of these results suggests the possibility of enzymatic hydrolysis at high solids to make the overall conversion cost-competitive., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Given that the glycerol organosolv pretreatment (GOP) can effectively improve the hydrolyzability of various lignocellulosic substrates, physicochemical changes of the substrate before and after the pretreatment was characterized to elucidate what is responsible for it. The effect of GOP on the main components and hydrolyzability of wheat straw was revisited. Results demonstrate that the GOP should be a promising candidate for the current pretreatment. Then the composition and structure of substrates was measured at multi-dimensional scales by using various analytic equipment such as TGA, SEM, AFM, CLSM, FT-IR, XRD and solid-state CP/MAS (13)C NMR. This paper reports some new insights on the mechanism behind that, which can be beneficial for further development, optimization, and scale-up of the GOP process., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Steam explosion and steam pre-treatment have proved capable of enhancing enzymatic saccharification of lignocellulosic materials. However, until now, these methods had not been compared under the same operational conditions and using the same raw material. Both pre-treatments lead to increased yields in the saccharification of Eucalyptus globulus; but results have been better with steam pre-treatments, despite the more accessible surface of exploded samples. The reason for this finding could be enzymatic inhibition: steam explosion causes a more extensive extraction of hemicelluloses and releases a greater amount of degradation products which can inhibit enzymatic action. Enzymatic inhibition is also dependent on the amount and chemical structure of lignin, which was also a contributing factor to the lower enzymatic yields obtained with the most severe pre-treatment. Thus, the highest yields (46.7% glucose and 73.4% xylose yields) were obtained after two cycle of steam treatment, of 5 and 3 min, at 183°C., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In this study, cellulose-rich fractions from bamboo were prepared with steam explosion pretreatment (SEP) followed by a successive alkaline delignification to improve the enzymatic digestibility for an efficient bioethanol production. The cellulose-rich fractions obtained were characterized by FT-IR, XRD, CP/MAS (13)C NMR, SEM, and BET surface area. It was found that the SEP alone significantly removed partial hemicelluloses, while the synergistic treatment by SEP and alkaline delignification removed most hemicelluloses and lignin. Results from enzymatic hydrolysis showed that SEP alone improved the enzymatic hydrolysis rate by 7.9-33.1%, while the synergistic treatment by SEP and alkaline delignification enhanced the rate by 45.7-63.9%. The synergistic treatment by SEP at 2.0 MPa for 5 min with water impregnation followed by a successive alkaline delignification with 0.5% NaOH and 70% ethanol containing 1.5% NaOH resulted in a maximum enzymatic hydrolysis rate of 70.6%., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Sulfite pretreatment to overcome the recalcitrance of lignocelluloses (SPORL) was applied to an empty fruit bunches (EFB) for ethanol production. SPORL facilitated delignification through lignin sulfonation and dissolution of xylan to result in a highly digestible substrate. The pretreated whole slurry was enzymatically saccharified at a solids loading of 18% using a relatively low cellulase loading of 15 FPU/g glucan and simultaneously fermented without detoxification using Saccharomyces cerevisiae of YRH400. An ethanol yield of 217 L/tonne EFB was achieved at titer of 32 g/L. Compared with literature studies, SPORL produced high ethanol yield and titer with much lower cellulase loading without detoxification., (Published by Elsevier Ltd.)

Bamboo was non-isothermally pretreated with hot water at 140-200°C for different times (10-120 min). The effects of pretreatment conditions on the degradation of carbohydrates, cellulose crystallinity, partial removal/relocation of lignin, morphologic change of the feedstock, and glucose yield during the enzymatic hydrolysis were investigated. The effective removal of amorphous cellulose and hemicelluloses led to the increase of crystalline index of the residues. In comparison with the raw material, the surface of the pretreated samples was irregular and numerous lignin droplets appeared on the cellulose bundle surface under the intense pretreatment conditions. The glucose conversion increased with the raise of pretreatment temperature and the prolongation of time, and the maximum conversion of 75.7% was achieved for the sample pretreated at 200°C for 120 min, whereas the untreated sample was only 15.7%. The result illustrated that hydrothermal pretreatment affected the composition of bamboo, and remarkably enhanced the enzymatic hydrolysis efficiency., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Most biomass pretreatment processes for sugar production are run at low-solid concentration (<10 wt.%). Subcritical carbon dioxide (CO2) could provide a more sustainable pretreatment medium while using relative high-solid contents (15 wt.%). The effects of subcritical CO2 pretreatment of sugarcane bagasse to the solid and glucan recoveries at different pretreatment conditions were investigated. Subsequently, enzymatic hydrolysis at different hydrolysis time was applied to obtain maximal glucose yield, which can be used for ethanol fermentation. The maximum glucose yield in enzyme hydrolyzate reached 38.5 g based on 100g raw material after 72 h of enzymatic hydrolysis, representing 93.0% glucose in sugarcane bagasse. The enhanced digestibilities of subcritical CO2 pretreated sugarcane bagasse were due to the removal of hemicellulose, which were confirmed by XRD, FTIR, SEM, and TGA analyses., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The commercial pre-hydrolysis kraft-based dissolving pulp production process can be a typical example for the demonstration/implementation of the integrated forest biorefinery concept. In this study, the concept of cellulase treatment of this dissolving pulp for enhancement of accessibility/reactivity in terms of viscose rayon production was demonstrated. The cellulase treatment resulted in the formation of additional openings/surface areas in the fiber structure via the possible action of "etching". As a result, the pore volume of pulp fibers increased, which led to the increase in the accessibility to xanthation, and thus Fock reactivity. Results showed that the cellulase treatment was effective in increasing the Fock reactivity, at a cellulase dosage of 2u/g (based on the dry weight of pulp), the Fock reactivity increased from 47.67% to 79.9%. The adoption of cellulase treatment to hardwood kraft-based dissolving pulp can provide an efficient approach for enhancing its performance in the commercial viscose-rayon process., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The present work investigated the effects of subcritical CO2 pretreatment of sugarcane bagasse at different CO2 pressure, pretreatment time, and temperature with relative high-solid concentration (15% w/v) to the composition of prehydrolyzate and the enzymatic hydrolysis. The results indicated that the maximum xylose yields in prehydrolyzate liquid were 15.78 g (combined 3.16 g xylose and 12.62 g xylo-oligosaccharides per 100g raw material). Due to the effective removal of hemicellulose, the maximum glucose yield in enzyme hydrolyzate reached 37.99 g per 100g raw material, representing 91.87% of glucose in the sugarcane bagasse. The maximal total sugars yield (combined xylose and glucose both in prehydrolyzate and enzymatic hydrolyzate) were 52.95 g based on 100g raw material. These results indicated that subcritical CO2 pretreatment can effectively improve the enzymatic hydrolysis, so it could be successfully applied to sugarcane bagasse., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Four fractions of one commercial sodium lignosulfonate (SXP) with different molecular weight (MW) and anionic polymers were studied to reduce non-productive adsorption of cellulase on bound lignin in a lignocellulosic substrate. SXP with higher MW had stronger blocking effect on non-productive adsorption of a commercial Trichoderma reesi cellulase cocktail (CTec2) on lignin measured by quartz crystal microgravimetry with dissipation monitoring. Linear anionic aromatic polymers have strong blocking effect, but they would also reduce CTec2 adsorption on cellulose to decrease the enzymatic activity. The copolymer of lignin and polyethylene glycol (AL-PEG1000) has strong enhancement in enzymatic hydrolysis of lignocelluloses, because it not only improves the cellulase activity to cellulose, but also blocks the non-productive cellulase adsorption on lignin. Apart from improving the cellulase activity to cellulose, the enhancements of enzymatic hydrolysis of lignocellulose by adding AL-PEG1000 and SXPs are the result of the decreased cellulase non-productive adsorption on lignin., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Rice straw is one of the most abundant agricultural residues in China. It is considered as a promising raw material for bioethanol production. In this work, rice straw was pretreated by sodium sulfite-formaldehyde (SF) for improving enzymatic saccharification. The SF pretreatment, using proven technology and industrialized equipment, showed efficient delignification selectivity and high carbohydrates retention in pretreated solid. The highest sugar yields of 79.0%, 88.8% and 71.1% for total sugar, glucan and xylan, respectively were obtained at an enzyme loading of 40 FPU/g-substrate after the raw material pretreated with 12% sodium sulfite at 160°C. About 75% of lignin was dissolved in pretreatment spent liquor and 78% of silica was retained in the residue of enzymatic hydrolysis. The results proved sulfite-formaldehyde as a promising pretreatment for the production of bioethanol as well as potential high value added by-products of silica nanoparticles and lignosulfonate., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

In this work, mixed hardwoods were pretreated by sodium sulfite-formaldehyde (SF). The effects of SF pretreatment on the chemical compositions and enzymatic hydrolysis of mixed hardwoods were investigated. SF pretreatment temperature had a significant effect on pulp yield and delignification, resulting in an increased efficiency of enzymatic hydrolysis. After 96 h of enzymatic hydrolysis at the cellulase loading of 40 FPU/g substrate, the yields of glucan and xylan on the basis of original wood were 37% and 11% for the pulp produced with 12% sulfite charge at 170 °C for 2 h. The total sugar recovery based on the sugar in original wood was 74%. These results indicate that sulfite-formaldehyde cooking is of great potential to be a pretreatment method for a greenfield mill to produce fuel ethanol from hardwood., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Pretreatment of rice straw by using renewable cholinium lysine ionic liquid ([Ch][Lys] IL)-water mixtures and subsequent enzymatic hydrolysis of the residues were conducted in this work. There is a clear correlation between the delignification capacity of the pretreatment solvent and its basicity. After pretreatment, surface area and pore volume of rice straw increased significantly, which substantially improved polysaccharides accessibility to enzymes and thus enhanced polysaccharides digestion. By carefully controlling the pretreatment severity (IL content, temperature and duration), loss of readily extractable xylan could be minimized. The sugar yields of 81% for glucose and 48% for xylose were achieved in the enzymatic hydrolysis of rice straw after pretreatment with 20% [Ch][Lys]-water mixture at 90 °C for 1 h. This pretreatment process is highly promising for industrial application because of high sugar yields, low energy input, short pretreatment time, and being environmentally benign and highly tolerant to moisture., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Filamentous fungi are widely used to produce cellulase, but how the fermentation conditions affect their production is not well known. In this study, cellulase production by Trichoderma viride in submerged fermentations with free cells in a stirred-tank reactor (STR) and immobilized cells in a rotating fibrous-bed bioreactor (RFBB) were investigated. Compared to free-cell fermentation, immobilized-cell fermentation gave 35.5% higher FPase activity and 69.7% higher saccharification yield of sugarcane bagasse (SCB). The secretory proteins in the fermentation broths were analyzed with two-dimensional gel electrophoresis (2-DE) and MALDI-TOF-TOF mass spectrometry, which identified 24 protein spots with differential expression levels. Among them, cellobiohydrolase CBH II and endoglucanase EG II were highly expressed and secreted in the immobilized-cell fermentation, while the free-cell fermentation produced more CBH І and EG ІV. These results showed that immobilized-cell fermentation with T. viride in the RFBB was advantageous for cellulase production., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Combination of liquid hot water pretreatment (LHWP) and wet disk milling (WDM) was investigated in this study to enhance the sugar recovery yield both in prehydrolyzate and enzymatic hydrolyzate. The results show that WDM with LHWP at 180 °C for 20 min produced maximum xylose and glucose yields of 91.62% and 88.12%, respectively, which are higher than that of dilute acid pretreatment or individual LHWP. Corresponding concentration of fermentation inhibitors such as acetic acid, HMF, and furfural in the prehydrolyzate are about 0.98, 0.07 and 0.78 g/L, respectively, which indicated that the detoxification may be not required in the next fermentation step. The acid-insoluble lignin recovery in the insoluble solid resulting from enzymatic hydrolysis, was 25.67/100g raw material, representing 90.7% of acid-insoluble lignin in the eucalyptus biomass. It can be concluded that liquid hot water pretreatment combined with wet disk milling can be successfully applied to eucalyptus., (Copyright © 2012. Published by Elsevier Ltd.)

Pseudo-lignin, which can be broadly defined as aromatic material that yields a positive Klason lignin value and is not derived from native lignin, has been recently reported to form during the dilute acid pretreatment of poplar holocellulose. To investigate the chemistry of pseudo-lignin formation, GPC, FT-IR and 13C NMR were utilized to characterize pseudo-lignin extracted from dilute-acid pretreated α-cellulose and holocellulose. The results showed that pseudo-lignin consisting of carbonyl, carboxylic, aromatic and aliphatic structures was produced from dilute acid pretreated cellulose and hemicellulose. Pseudo-lignin extracted from holocellulose pretreated at different conditions had similar molecular weights (Mn∼1000 g/mol; Mw∼5000 g/mol) and structural features (carbonyl, carboxylic, aromatic and methoxy structures). These characterizations have provided the pseudo-lignin formation mechanisms during pretreatment. The presence and structure of pseudo-lignin is important since pseudo-lignin decreases the enzymatic conversion., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Hydrothermal pretreatment was performed on the leaves and internodes portions of Alamo switchgrass, Panicum virgatum L., to enhance the digestibility of cellulose towards cellulase. It was observed that extractives free leaves portion provided 18.1% lower pretreatment gravimetrical yield and 33.8% greater cellulose-to-glucose yield than internodes portion. The degree of polymerization (DP) and ultrastructure of cellulose were determined by gel-permeation chromatography and solid-state cross polarization/magic angle spinning (13)C NMR experiments. The results suggested that hydrothermal pretreatment hydrolyzed amorphous cellulose and yielded a product enriched in paracrystalline cellulose. Furthermore, the DP of cellulose was reduced to one third of the origin value after hydrothermal pretreatment. The resulting biomass after pretreatment for leaves and internodes has similar cellulose ultrastructure and chemical profiles. The results of the enzymatic hydrolysis studies of cellulose suggest that the reduced DP of cellulose of pretreated switchgrass was an important factor influencing the enhanced digestibility of pretreated switchgrass., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

This study demonstrated the efficiency of Ca(II) and Mg(II) in removing inhibition of enzymatic hydrolysis by lignosulfonate through non-productive adsorption of enzymes. Adding 1 mmol/g cellulose of either metal salt restores approximately 65% of the activity lost when a pure cellulose/cellulase solution is spiked with lignosulfonate. Addition of either Ca(II) or Mg(II) is also effective in counteracting soluble inhibitors of cellulase present in unwashed aspen solid substrate produced by SPORL (sulfite pretreatment to overcome recalcitrance of lignocelluloses). Soluble inhibitors are often removed by thoroughly washing the lignocellulosic solid substrate following pretreatment. It was determined that adding 1 mmol of MgSO(4)/g substrate (oven dry) to the unwashed aspen substrate gave enzymatic substrate digestibility (SED) equivalent to that of washing for a range of enzyme loadings. These results demonstrate that applying divalent metal salts eliminates the need for washing, thereby saving considerable process water and cost for production of chemicals and biofuels from lignocellulose., (Published by Elsevier Ltd.)

Under the current energy scenario, the development of alternatives to fossil fuels, like bioethanol from lignocellulosic materials, is highly relevant. Therefore it is important to search and study new raw materials and to optimize the different steps that lead to bioethanol production. In this work, acid diluted pretreatment was optimized considering the release of sugars. Under the optimal conditions, the reducing sugars yield was of 293.4mg/g of dry biomass in liquid fraction. The tested pretreated samples of Pterospartum tridentatum that presented a higher glucose yield in enzymatic saccharification where those that were subject to a pretreatment at 180°C for 75min with 2.75% (w/w) of sulfuric acid when using a biomass/liquid ratio of 2.25g/10mL leading to a maximum yield of glucose that was 92% of the theoretical maximum. From the fermentation of filtrates it was possible to obtain a maximum ethanol yield of 0.26g ethanol/g total sugars, without previous detoxification., (Copyright © 2010. Published by Elsevier Ltd.)

In this study, we examined the effect of oxygen delignification on the rate and extent of enzymatic hydrolysis (using commercial cellulase and beta-glucosidase) of a number of lignocellulosic substrates, including kraft pulp (model substrate), pulp mill primary clarifier sludge (PCS) and steam-exploded Douglas fir chips. Oxygen delignification removed up to 67% of the lignin from softwood pulp and improved the rate of, and yield from, hydrolysis by up to 111% and 174%, respectively. Glucose yield varied linearly with fractional lignin removal. Oxygen delignification of primary clarifier sludge improved hydrolysis yield by up to 90%. However steam-exploded Douglas fir was very resistant to hydrolysis at low enzyme loading, and oxygen delignification decreased hydrolysis rate and yield.