Michel Petit-Conil

Fusion of a family 1 carbohydrate binding module of Aspergillus niger to the Pycnoporus cinnabarinus laccase for efficient softwood kraft pulp biobleaching.

Pycnoporus cinnabarinus laccase was fused to the C-terminal linker and carbohydrate binding module (CBM) of Aspergillus niger cellobiohydrolase B (CBHB). The chimeric enzyme of molecular mass 100 kDa was successfully produced in A. niger. Laccase-CBM was further purified to determine its main biochemical properties. The Michaelis-Menten constant and pH activity profile were not modified, but the chimeric enzyme was less thermostable than either the P. cinnabarinus laccase or the recombinant laccase produced in the same strain. Laccase-CBM was able to bind to a cellulosic substrate and, to a greater extent, to softwood kraft pulp. Binding to the pulp was shown to be mainly time and temperature-dependent. Laccase-CBM was further investigated for its softwood kraft pulp biobleaching potential and compared with the P. cinnabarinus laccase. Addition of a CBM was shown to greatly improve the delignification capabilities of the laccase in the presence of 1-hydroxybenzotriazole (HBT). In addition, ClO(2) reduction using 5 U of chimeric enzyme per gram of pulp was almost double than that observed using 20 U of P. cinnabarinus laccase per gram of pulp. We demonstrated that conferring a carbohydrate binding capability to the laccase could significantly enhance its biobleaching properties.

Green synthesis of xylan hemicellulose esters

The esterification of xylan type hemicelluloses, isolated from birchwood, was carried out firstly in homogeneous conditions using N,N-dimethylformamide (DMF) and lithium chloride (LiCl) in the presence of 4-dimethylaminipyridine (DMAP). The degree of substitution (DS) of xylan acetates ranged between 0.9 and 2.0 as a function of experimental conditions. Due to the problems of toxicity and recycling of DMF, an alternative method of esterification is reported in the second part of this work, performing in the absence of organic solvent and using DMAP or methanesulfonic acid (MSA) as catalysts. Acetylation reaction catalyzed by MSA was developed through an experimental design in order to achieve the highest DS under the mildest conditions. The significant factors and their interactions were identified. The optimization of reaction parameters allowed to obtain a high DS (1.6) and maximal yield (85%). Moreover, the reactivity of propionic and hexanoic anhydrides was evaluated and hydrophobic xylan esters with low degrees of substitution were obtained.

Green nondegrading approach to alkyne-functionalized cellulose fibers and biohybrids thereof: synthesis and mapping of the derivatization.

Alkyne-functionalized cellulose fibers have been generated through etherification under basic water or hydroalcoholic conditions (NaOH/H(2)O/isopropanol). For a given NaOH content, the medium of reaction and, more particularly, the water/IPA ratio, were shown to be of crucial importance to derivatize the fibers without altering their integrity and their crystalline nature. It was shown that the degree of substitution (DS) of the fibers increases concomitantly with isopropanol weight ratio and that, contrary to water or water-rich conditions, derivatization of fibers under isopropanol-rich conditions induces an alteration of the fibers. Optimization of etherification conditions in aqueous media afforded functionalized cellulose materials with DS up to 0.20. Raman confocal microscopy on derivatized fibers cross sections stressed that alkyne moieties are incorporated all over the fibers. The resulting fibers were postfunctionalized by molecular probes and macromolecules in aqueous or water-rich conditions. The effectiveness of the grafting was strongly impacted by the nature of the coupling agents.

Scale deposits in kraft pulp bleach plants with reduced water consumption: A review

The general tendency in the pulp industry towards reduced fresh water consumption and minimum effluent causes major deposit problems in mills. Chemical pulp bleach plants are affected by several types of mineral deposits, the most frequent being calcite, barite and calcium oxalate. In this review, the phenomena leading to scaling in chemical pulp bleaching are discussed, together with strategies for limiting deposits. The merits of various chemical methods in estimating scaling risks are compared. Chemical speciation methods are used throughout this review to gain a better understanding and prediction of scaling phenomena. Coupled chemical process simulations are anticipated to be a crucial way of solving deposition problems in bleach plants.

Hydrophobic properties conferred to Kraft pulp by a laccase-catalysed treatment with lauryl gallate.

Hydrophobic properties were conferred to a high-lignin-content Kraft pulp by a laccase-catalysed treatment in the presence of lauryl gallate (LG). The treatment resulted in a two-fold increase in contact angle and conferred water absorption resistance to the pulp. Kappa number was increased, indicating that some phenolic compounds were incorporated in the pulp. A control treatment with LG alone did not affect water absorption, demonstrating that laccase was essential to attain these new properties. The loss of hydrophobicity after an acetone Soxhlet extraction highlighted that adsorbed acetone-soluble compounds played a key role in the properties. GC-FID and HPSEC-UV analysis of the acetone extract indicated the formation of dodecanol and different phenolic oligomers. SEM images showed the treatment-induced changes in the fibre network. Additional experiments with various reaction times and reactant concentrations highlighted the role of LG oxidation products in the introduction of absorption resistance.

Periodate oxidation of 4-O-methylglucuronoxylans: Influence of the reaction conditions.

This work aims at studying the sodium periodate oxidation of 4-O-methylglucuronoxylans (MGX) in different experimental conditions for a control of the oxidation degree. A series of sodium periodate oxidation reactions were conducted at three NaIO4/xylose molar ratios: 0.05, 0.20 and 1.00. The effects of xylan molar mass, xylan concentration and reaction temperature on the reaction rate have been evaluated by UV/visible spectroscopy at 0.20 NaIO4/xylose ratio. No depolymerization is observed at 0.05 ratio while depolymerization occurs at 0.20 and is even complete at 1.00 NaIO4/xylose ratio. An increase of the reaction temperature - up to 80 °C - leads to an increase of the oxidation rate with no effect on the depolymerization. At high xylan concentrations, the oxidation rate increases but promotes chains aggregation.

Biohybrid cellulose fibers: Toward paper materials with wet strength properties

Herein, we report on the preparation of novel cellulose-PEG biohybrid papers with wet strength properties. The biohybrid paper sheets are obtained using a two-step procedure where ω- or α, ω-azide functionalized PEG chains are anchored onto alkyne-functionalized wood fibers through CuAAC ligation in mild and aqueous conditions. The incorporation of the PEG grafts mostly occurs at the periphery of the cellulose fibers and degrees of substitution up to 0.028 are obtained. The presence of PEG grafts significantly increases the tensile, burst and tear strength properties in the wet state, the reinforcement being more pronounced for fibers grafted with α,ω-azide PEG. This reinforcement is consistent with a relatively sparse hetero-crosslink reaction creating inter-fiber covalent bonds and forming a cellulose network within the cell wall.

Assessing cypermethrin penetration in Pinus sylvestris wood products by immuno-electron microscopy

The preservative efficacy of organic biocidal products is strongly related to their capacity of penetration and retention within wood tissues. The specific detection of the pyrethroid insecticide cypermethrin is currently obtained after extraction followed by chemical analysis by chromatography techniques. However, visualizing the insecticide molecule within the wood structure requires specific probes together with microscopy techniques. Therefore, the aim of the present work was to prepare a polyclonal antibody directed against cypermethrin and to implement it on Pinus sylvestris wood samples treated with technical cypermethrin. The antibody was tested on cypermethrin-impregnated wood, and the specific recognition of the insecticide was visualized by transmission electron microscopy (TEM). The immunogold-TEM assay evidenced the capacity of the synthetic biocide active substance to penetrate in the wood. The depth of penetration was measured on sections taken at increasing distances from the coated surface of the wood. Under the present conditions of application, cypermethrin was shown to penetrate up to the 6- to 9-mm zone below the surface. Such results correlated with chemical analyses carried out by GC-ECD after extraction. In addition, the immuno-TEM investigation allowed visualizing, for the first time at the ultrastructure scale of resolution, that cypermethrin was able to diffuse within the secondary wood cell walls. The scarce labeling of the compound middle lamella shows that the chemical does not diffuse freely in this part of the cell walls. The results suggest that the adsorption within the cell walls is an essential factor for the retention of cypermethrin and for its permanence in treated wood products.

Qualité bioénergétique de la biomasse forestière en vue de la production de biocarburants de deuxième génération

Second-generation liquid fuels are now at the development stage that precedes their industrial implementation. They offer significant market opportunities for French lignocellulosic resources both from forests and crops. Those resources are limited and it is important to avoid competing against food crops or destabilising the sectors that already operate. It is therefore essential for the actors in research on these resources and processes to work in partnership and produce a common response to these issues. The REGIX project has provided an assessment of the suitability of various types of forests and cultivated biomass to the needs of processes for producing second-generation biofuels via biochemical and thermochemical processes. This article provides an overview of this approach for four classes of forest biomass: short and very short rotation coppice (SRC and VSRC), wood chips from hardwoods and from softwoods. The results show that these classes are very well-suited to most of the specifications for these processes, but a number of drawbacks were also noted, in particular for VSRC.