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This study reports the cellulo-xylanolytic cocktail production from Hypocrea lixii GGRK4 using multi-objective genetic algorithm-artificial neural network tool, resulting in 8.32 ± 1.07 IU/mL, 51.53 ± 3.78 IU/mL activity of CMCase and xylanase, respectively with more than 85 % residual activity at 60 °C and pH 6.0. Interestingly, metal ions viz. K + and Ca 2+ stimulated the enzyme activity, whereas Fe 2+ and Cu 2+ reduced the activity. Significant amounts of hydrophobic compounds, chromophores, and phenolics were released after wastepapers deinking. The deinking efficiency of 73.60 ± 2.45 % and 38.60 ± 1.34 % was obtained for photocopier paper and newspaper, respectively, whereas brightness of 89.90 ± 2.10 % ISO and 44.90 ± 1.63 % ISO was reported for both types of waste papers. The physical strength of deinked photocopier paper and newspapers, i.e., tensile index (3.10 and 0.50 %), tearing index (7.10 and 4.83 %), and burst factor (8.61) were enhanced whereas double fold property was decreased proving wastepaper reusability. This consortium showed effective and significant enzymatic deinking efficiency for recycled wastepapers., 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 © 2024 Elsevier Ltd. All rights reserved.)

In this work, laccase treatment was employed to enhance nanofiltration process by lignin removal. Results showed that the membrane filterability was increased in terms of deionized water flux and PHL filtration process. On the other hand, the hemicellulosic sugars were negligible affected and can be concentrated to 172 g/L, which was increased about 300% from the original one. The combined laccase-nanofiltration process provides an alternative approach to utilize hemicellulosic sugars of PHL in an environmentally friendly way., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

In present study, xylanase and laccase were produced in a cost-effective manner up to 10 kg substrate level and evaluated in elemental chlorine free bleaching of Eucalyptus kraft pulp. Compared to the pulp pre-bleached with xylanase (15%) or laccase (25%) individually, the ClO2 savings were higher with sequential treatment of xylanase followed by laccase (35%) at laboratory scale. The sequential enzyme treatment when applied at pilot scale (50 kg pulp), resulted in improved pulp properties (50% reduced post color number, 15.71% increased tear index) and reduced AOX levels (34%) in bleach effluents. The decreased AOX level in effluents will help to meet AOX discharge limits, while improved pulp properties will be value addition to the paper., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Lignin removal is essential for value-added utilization of hemicelluloses and acetic acid present in the prehydrolysis liquor (PHL) of a kraft-based hardwood dissolving pulp production. In this paper, a novel process concept, consisting of laccase-induced lignin polymerization, followed by filtration/flocculation, was developed to enhance the lignin removal. The results showed that the lignin removal increased from 11% to 46-61% at laccase concentration of 1-4 U mL(-1). The GPC results showed that the molecular weight of the lignin from the laccase treated PHL was increased by 160% in comparison with the original one. The subsequent flocculation using singular Poly-DADMAC system or dual polymer system of Poly-DADMAC/CPAM can further remove 10-15% lignin. The concentrations of hemicelluloses and acetic acid were negligibly affected during the laccase treatment, while flocculation caused 12-15% of total sugar loss. Additionally, the process incorporates this new concept into the kraft-based dissolving pulp production process was proposed., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Bleached cellulose with good end-properties (≈ 90% ISO brightness and 62% cellulose preservation) was obtained by using a totally chlorine-free biobleaching process (TCF). Unbleached sulphite cellulose was treated with Trametes villosa laccase in combination with violuric acid. This enzymatic stage (L) was followed by a chelating stage (Q) and then by a hydrogen peroxide stage reinforced with pressurized oxygen (Po), resulting to an overall LQPo sequence. The use of violuric acid was dictated by the results of a preliminary study, where the bleaching efficiency of various natural (syringaldehyde and p-coumaric acid) and synthetic mediators (violuric acid and 1-hydroxybenzotriazole) were assessed. The outstanding results obtained with laccase-violuric acid system fulfil most of the characteristics of commercial dissolving pulp, totally acceptable for viscose manufacturing or CMC derivatives, with the added advantage that the enzymatic treatment saved 2h of reaction time and about 70% of hydrogen peroxide consumption, relative to a conventional sequence (Po)., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

A new approach for the hydrophobization of finished cellulosic substrates based on a previously reported enzymatic technique is proposed. Commercial finished paper was hydrophobized by using laccase from Trametes villosa in combination with lauryl gallate (LG) as hydrophobic compound. The efficiency of the method was increased by the use of a lignosulfonate as a natural dispersant to improve the surface distribution of LG on the paper, raise its hydrophobicity and help preserve the enzyme activity. No similar threefold effect from a single compound for the improvement of enzymatic treatments was previously reported. The influence of processing conditions including the LG dose, treatment time and temperature was also examined, resulting in further increased hydrophobicity. Efficient fiber bonding and chemical functionalization were confirmed by thorough washing and Soxhlet extraction of the paper. As shown here for the first time, enzyme treatments have the potential to improve the surface hydrophobicity of paper-based media., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Pycnoporus cinnabarinus laccase and a chimeric laccase-CBM were applied in softwood kraft pulp biobleaching in the presence of 1-hydroxybenzotriazole (HBT). The presence of CBM could enhance the laccase biobleaching potential as a decrease in the enzymatic charge and chlorine dioxide consumption, as well as an increase in pulp brightness were observed. Laccase/HBT treatment could be improved by increasing oxygen pressure from 1 to 3bar and pulp consistency from 5% to 10%. Conversely, under the same conditions, no improvement of laccase-CBM/HBT treatment was observed, indicating a different behavior of both systems. However, laccase-CBM/HBT treatment led to a better preservation of pulp properties. This effect was probably due to fiber surface modifications involving the action of the CBM. Transmission electron microscopy examination of pulp fibers indicated a retention of laccase-CBM inside the pulp fibers due to CBM binding and an increased external microfibrillation of the fibers due to enzymatic treatments., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Nine phenol derivatives, p-coumaric acid (PC), vanillin (V), acetovanillone (AV), acetosyringone (AS), syringaldehyde (SA), coniferaldehyde (CLD), ferulic acid (FRC), sinapic acid (SNC), and sinapyl aldehyde (SLD) were assayed as laccase redox mediators in the biobleaching of kenaf and sisal pulps. As a general behaviour, the phenolic mediators increased the kappa number (KN) and reduced the brightness of pulps. In particular, these changes were found to depend in a linear manner on the energy of the highest occupied molecular orbital (E(HOMO)) of the mediators. The phenolic mediator with the lowest E(HOMO) (PC) led to the highest increase of KN and the lowest reduction of brightness. On the contrary, syringyl derivatives (i.e. SA) with high E(HOMO) values caused small KN increases and significant losses of brightness. This behaviour was explained on the basis of a competition between grafting and polymerisation processes. The former basically affects KN, whereas the latter affects pulp brightness., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Modification of the physical properties of old corrugated container (OCC) pulp with laccase or a laccase-mediator (ABTS, HBT, VA) system was investigated under select enzymatic concentrations and reaction times. The optimal conditions for laccase treatment shown to be using a laccase dose of 160U/g o.d. pulp, a treatment time of 20h at 25°C, pH 7 with a pulp consistency of 5%. Results showed that the Lac-HBT treated OCC pulp gave the best strength properties, improving tensile strength by 15.7%. The increase in the carboxyl group content of OCC laccase or Lac-HBT treated pulp led to the increase in the swelling ability and bonding between fibers. Microscope images showed the fiber surface became rougher and more collapsible after Lac-HBT treatment. FT-IR data showed that new carboxylic acid groups were formed during Lac-HBT treatment., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The environmental impact of enzyme bleaching stages applied to oxygen-delignified eucalypt kraft pulp was assessed via the chemical oxygen demand (COD), color, absorbance spectrum, residual enzyme activity and Microtox toxicity of the effluents from a laccase-HBT (1-hydoxybenzotriazole) treatment. The influence of the laccase and HBT doses, and reaction time, on these effluent properties was also examined. The laccase dose was found to be the individual variable most strongly affecting COD, whereas the oxidized form of HBT was the main source of increased color and toxicity in the effluents. Moreover, it inactivated the enzyme. Oxidation of the mediator was very fast and essentially dependent on the laccase dose. Using the laccase-mediator treatment after a xylanase stage improved pulp properties without affecting effluent properties. This result holds great promise with a view to the industrial implementation of biobleaching sequences involving the two enzymes in the future., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Thermogravimetric analyses (TGA) was applied to study the effects of enzymatic bleaching of eucalyptus pulp with xylanase and a laccase-mediator system. The thermal degradation profile of the pulps was sensitive to the enzymatic treatments. Xylanase treatment produced an ordered and clean microfibril, whereas laccase oxidized surface cellulose chains and increased the amorphous (paracrystalline) cellulose content. In this case, pulp viscosity decreased from 972 to 859 mL/g and apparent pulp crystallinity calculated from TGA data decreased almost 50%. Alkaline extraction was necessary to recover pulp crystallinity and to remove oxidized lignin in the laccase-treated samples. TGA data allowed differentiating and quantifying crystalline and amorphous cellulose. This thermogravimetric approach is a simple method in order to monitor superficial changes in cellulosic microfibrils., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Functionalization of sisal specialty pulp fibers by laccase-catalysed grafting of ferulic acid (FRC) was investigated. To this end, the extent of phenol coupling to fibers under different reaction conditions (laccase and FRC rates, and time) was evaluated in terms of pulp properties including kappa number (expressed as the combined contributions of lignin and hexenuronic acids), brightness, Klason lignin and surface anionic charge after Soxhlet extraction of acetone-treated pulp. The specific treatment resulting in the highest degree of grafting was then used in a comparative study of the effects of applying the laccase-FRC system to refined and unrefined pulp with a view to confirming whether the increased surface area obtained by effect of fibrillation would lead to enhanced grafting. Based on the results, refining the pulp prior to the enzyme treatment resulted in increased grafting which in turn led to handsheets with improved strength-related properties (particularly wet tensile strength) relative to control samples., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The effect of a pretreatment consisting of steam explosion (SE) followed by a laccase mediator system (LMS) stage on Eucalyptus globulus kraft pulping has been evaluated and compared with fungal pretreatments. Pretreatment with SE and LMS was more efficient than pretreatments using Pycnoporus sanguineus and Trametes sp. I-62. Steam explosion not only improved the enzyme penetration into the wood chips and shortened the pulping process by 60%, but also extracted around 50% of the hemicelluloses which could be converted into value-added products. The optimal conditions for the LMS treatment were 3h, 3UA/g and 40°C. Compared to SE, the SE/LMS treatment yielded an increase in delignification of 13.9% without affecting pulp properties, provided a similar screened kraft yield, and reduced consumption of chemical reagents Na(2)S and NaOH by 11.5% and 6.3%, respectively. Therefore, SE/LMS is a promising pretreatment for converting the pulp mill into a forest bio-refinery., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The ability of two natural phenols to act as mediators of the recombinant Myceliophthora thermophila laccase (MtL) in eucalypt-pulp delignification was investigated. After alkaline peroxide extraction, the properties of the enzymatically-treated pulps improved with respect to the control. The pulp brightness increased (3.1 points) after the enzymatic treatment with MtL alone, but the highest improvements were obtained after the MtL treatment using syringaldehyde (4.7 points) and especially methyl syringate (8.3 points) as mediators. Likewise, a decrease in kappa number up to 2.7 points was obtained after the MtL-methyl syringate treatment, followed by decreases of 1.4 and 0.9 points after the treatments with MtL-syringaldehyde and MtL alone, respectively. On the other hand, removal of the main lipophilic extractives present in eucalypt pulp was observed after the above laccase-mediator treatments. Finally, the doses of both MtL and methyl syringate were reduced, and results compatible with industrial implementation were obtained., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The effects of 1-hydroxybenzotriazole (HBT) and the natural laccase mediators gallic acid, caffeic acid and p-hydroxybenzoic acid, on the enzymatic bleaching of flax pulp were compared. The treatment was performed under atmospheric air and oxygen pressure, and, for the first time, monitored by thermogravimetric analysis (TGA) for comparison with chemical analysis, FTIR and X-ray diffraction (XRD) spectroscopies. Thermogravimetric data were simulated by applying a nucleation kinetic equation to a combustion model based on four pseudo-components (hemicellulose, amorphous and crystalline cellulose, and lignin). The results thus obtained show that the effects of the natural mediators are similar to those of HBT and lead to increased order in cellulose microfibril surfaces. An increase in pulp crystallinity was additionally exposed by the XRD and FTIR techniques, and a reduction in microfibril equatorial size by the XRD patterns. Simulated DTG curves were used to determine the kinetic parameters for thermal degradation., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Deinking of old newsprint (ONP) by combining hemicellulase with laccase-mediator system (LMS) was investigated, and surface chemical composition and fiber morphology changes during the deinking process were studied by electron spectroscopy for chemical analysis (ESCA), contact angle (CA), attenuated total reflectance fourier transform infrared spectrometry (ATR-FTIR), fiber quality analyzer (FQA), and environmental scanning electronic microscopy (ESEM). Results showed that, compared to the pulp deinked with hemicellulase or LMS individually, effective residual ink concentration (ERIC) was lower for the hemicellulase/LMS-deinked pulp. This indicated that there is a synergistic deinking effect between hemicellulase and LMS. It was found that O/C ratio of the fiber surface increased and the surface coverage of lignin decreased during the hemicellulase/LMS deinking process. The contact angle of the hemicellulase/LMS-deinked pulp was lower than that of pulps deinked with each individual enzyme. ESEM observations showed that more fibrils appeared on the fiber surface due to synergistic treatment., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Several effluents from laccase-mediator treatments of kraft pulp were recovered and subsequently reused with fresh pulp in order to simulate recirculation of effluents during biobleaching. The effluents were used as a new bleaching stage without any modification except enzyme addition. Pulp treated with effluents were afterwards chemically bleached by using the simple sequence LQPo, where L represents the treatment with effluent and laccase addition, Q is a chelating stage and Po is an alkaline peroxide stage. This system showed a promising potential on delignification, with kappa number ranging from 5.5 to 6.6 after LQPo sequence, depending on the type of effluent employed in L stage. Improvements on pulp brightness were also reported compared with control experiment., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

The aim of this work was to develop an innovative method for the internal sizing of paper by use of laccase and hydrophobic compounds. Nine different products containing hydrophobic moieties were tested in combination with laccase derived from Trametes villosa on Eucalyptus globulus kraft pulp in order to assess their internal sizing capability. The strongest internal sizing effect was obtained with lauryl gallate (LG). Heat treatment of the handsheets was found to increase the resistance to water absorption of internally sized samples significantly. Tests were conducted under variable operating conditions, including enzyme and reactant doses and treatment time. In addition to altering the water absorption rate, internal sizing with the laccase-LG treatments was found to affect the mechanical and optical properties of the handsheets. As shown in this work, treatments based on laccase and a hydrophobic compound (particularly lauryl gallate), can provide a new, effective biotechnological method for the internal sizing of paper., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Application of a laccase mediator system (an L stage) to TCF and ECF bleached pulp from Eucalyptus globulus with low residual lignin content (KN(lig)1.0) provides useful information about its effects on hexenuronic acids, functional groups (carboxyl and carbonyl) and electrokinetic properties such as zeta potential and surface charge. The use of laccase from Trametes villosa in combination with the mediator 1-hydroxybenzotriazole (HBT) was found to oxidize cellulose to carbonyl groups and reduce the amount of carboxyl groups present in TCF pulp by effect of its partially removing hexenuronic acids from it. This result may open up new prospects for improving brightness stability in pulp. In addition, the laccase mediator system modifies the surface charge and zeta potential in the fibre suspension for the removal of ionizables groups in TCF pulp. This result has no adverse effect on the pulp refining efficiency. L treatment requires less mechanical energy than conventionally refined pulp to obtain an optimal tensile and tear index in handsheets. This behavior may be attributable to the modification of the electrokinetic properties., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Modification of properties of old newspaper (ONP) deinked pulp with laccase and histidine was investigated. It was found that the optimum conditions for laccase-histidine treatment were: the concentration of laccase 0.9 U/g dry pulp, the concentration of histidine 1% relative to the dry pulp, room temperature, reaction time 1.5h, pH 7, the pulp consistency 5% and O(2) atmosphere. The results also showed that, in the optimum conditions, compared to the control pulp, the wet tensile strength, the carboxyl group content and water retention value of ONP treated with laccase and histidine were increased by 55.1%, 39.1% and 45.7%, respectively. Moreover, environmental scanning microscope images showed that more collapse and more fibrillation were observed on the laccase-histidine-treated fiber surface than the control samples, which led to form better bonding between fibers in handsheets resulting in the increase of the paper strength of laccase-histidine-treated ONP pulp., (Copyright 2010 Elsevier Ltd. All rights reserved.)

A new biobleaching sequence, with two enzymatic stages based on the application of laccase-mediator systems, was tested (L(1)EL(2)QPo) in order to increase the effectiveness of enzyme delignification on eucalypt kraft pulp. Different synthetic -1-hydroxybenzotriazole (HBT) and violuric acid (VA) - and natural - syringaldehyde (SyAl) - mediators were used in the laccase stages and the biobleached pulp were compared in terms of chemical, optical and physico-mechanical properties. The pulp bleached with HBT or VA showed similar delignification (64.1% and 65.9% respectively) and optical properties (86.4% and 86.1% ISO brightness respectively) than an industrial TCF pulp (68.3% delignification and 84.8% ISO brightness). SyAl improved these properties in a lower extent (56.71% delignification and 80.52% ISO brightness). Regarding physico-mechanical properties of pulp, the biobleaching sequence had no a negative effect, even some slight improvements were observed in very specific cases., (Copyright 2010 Elsevier Ltd. All rights reserved.)

High-quality flax pulp was bleached in a short totally-chlorine-free (TCF) sequence. The LP biobleaching sequence includes an enzyme treatment with laccase in the presence of HBT as mediator (L stage) and a hydrogen peroxide stage (P stage). The operating conditions for the laccase HBT system were optimized by using a sequential statistical plan involving four variables, the influence of which on the properties of the pulp after the P stage was examined over enclosed operation conditions regarding a future industrial application. Mathematical models accurately predicting both pulp properties in terms of the previous four variables. This biobleaching sequence allows obtaining an increase in ISO brightness of 40% and a delignification of 80%. Process variables optimization in flax pulp biobleaching allows establishing the application conditions most appropriated to obtain certain pulp properties and that turn out to be easily adaptable to existing industrial processes. As a novelty, toxicity measurements were determined in L stage effluents. The results of toxicity show that it is possible to apply this LP sequence without having a negative environmental impact.

The effects of laccase-natural mediator systems (LMS) on sisal pulp and their potential for either biobleaching or functionalizing (via radical-coupling) its fibres were investigated. The enzyme treatment (L stage) was followed by extraction with hydrogen peroxide in order to determine whether observable effects could be enhanced by removing LMS-modified lignin. Four different plant phenols [viz. the p-hydroxycinnamic compounds sinapic acid (SNC), ferulic acid (FRC), coniferyl aldehyde (CLD) and sinapyl aldehyde (SLD)] were used as laccase redox mediators and their effects on pulp and effluents compared with those of the synthetic compound 1-hydroxybenzotriazole (HBT). During the L stage performed with HBT, laccase underwent a loss of 99% and 78% of the initial activity, in the absence and presence of pulp, respectively. With natural mediators inactivation was markedly reduced, being the residual activity between 65% and 100% of the initial one, in the presence of pulp. The pulp was found to protect the enzyme against inactivation: the activity was only reduced by 45% in its presence. Under the operating conditions used the natural mediators proved less efficient than HBT in facilitating pulp bleaching; rather, they tended to bind to pulp fibres. This effect could be used to functionalize fibres in order to improve intrinsic properties of pulp or introducing novel ones (e.g. antimicrobial, antioxidant, optical properties, etc.). This paper shows for the first time the application of laccase-mediator systems to sisal pulp.

Different operating conditions (viz. pulp consistency, oxygen pressure and treatment time) in the biobleaching of eucalyptus kraft pulp with the laccase-HBT system was tested in order to describe their effect and normalize a biobleaching protocol. A high O(2) pressure (0.6MPa) was found to result in improved laccase-assisted delignification of the pulp. Also, a high pulp consistency (10%) and a short treatment time (2h) proved the best choices with a view to obtaining good pulp properties (kappa number and ISO brightness) under essentially mild conditions. The laccase-HBT treatment was found to result in slight delignification (in the form of a 20-27% decrease in kappa number); however, an alkaline extraction stage raised delignification to 41-45%, a much higher level than those obtained in the control tests (16-23%). Also, the use of hydrogen peroxide in the extraction stage resulted in improved brightness (14-19%), but in scarcely improved delignification (4-7%). Treating the pulp with the laccase-HBT system reduced the amount of hydrogen peroxide required for subsequent alkaline bleaching by a factor of 3-4 relative to control tests.

The natural phenolic compounds syringaldehyde and vanillin were compared to the synthetic mediators 1-hydroxybenzotriazole, violuric acid and promazine in terms of boosting efficiency in a laccase-assisted biobleaching of eucalyptus kraft pulp. Violuric acid and 1-hydroxybenzotriazole revealed to be the most effective mediators of the bioprocess. Nevertheless, laccase-syringaldehyde system also improved the final pulp properties (28% delignification and 63.5% ISO brightness) compared to the process without mediator (23% and 61.5% respectively), in addition to insignificant denaturation effect over laccase. The efficiency of the biobleaching process was further related to changes in non-conventionally used optical and chromatic parameters of pulp, such as (L*), chroma (C*) and dye removal index (DRI) showing good correlation. Adverse coupling reactions of the natural phenolic mediators on pulp lignin were predicted by electrochemical studies, demonstrating the complexity of the laccase-mediator reaction on pulp.

There has been increasing interest in extracellular enzymes from white rot fungi, such as lignin and manganese peroxidases, and laccases, due to their potential to degrade both highly toxic phenolic compounds and lignin. The optimum cultivation conditions for laccase production in semi-solid and liquid medium by Trametes versicolor, Trametes villosa, Lentinula edodes and Botrytis cinerea and the effects of laccase mediator system in E1 effluent were studied. The higher laccase activity (12756 U) was obtained in a liquid culture of T. versicolor in the presence of 1 mM of 2,5-xylidine and 0.4 mM copper salt as inducers. The effluent biotreatments were not efficient in decolorization with any fungal laccases studied. Maximum phenol reduction was approximately 23% in the absence of mediators from T. versicolor. The presence of 1-hydroxybenzotriazole did not increase phenol reduction. However, acetohydroxamic acid, which was not degraded by laccase, acted very efficiently on E1 effluent, reducing 70% and 73% of the total phenol and total organic carbon, respectively. Therefore, acetohydroxamic acid could be applied as a mediator for laccase bioremediation in E1 effluent.

The recent interest in the utilisation of agricultural fibres has promoted research into their potential as raw materials for the pulp and paper industry. In the current study, we report on the effect of biological pretreatment of wheat straw by Streptomyces cyaneus on the performance of the handsheets produced from the treated pulps. The pre-treatment of wheat straw with S. cyaneus had a positive effect on both the burst and tear indexes of the pulps but had a negative impact on tensile index. No significant variation in permeability and in folding endurance was observed. Manipulation of handsheets from wheat straw through biological treatment may therefore result in improved quality traits.