A GH115 alpha-glucuronidase from Schizophyllum commune contributes to the synergistic enzymatic deconstruction of softwood glucuronoarabinoxylan

Background: Lignocellulosic biomass from softwood represents a valuable resource for the production of biofuels and bio-based materials as alternatives to traditional pulp and paper products. Hemicelluloses constitute an extremely heterogeneous fraction of the plant cell wall, as their molecular structures involve multiple monosaccharide components, glycosidic linkages, and decoration patterns. The complete enzymatic hydrolysis of wood hemicelluloses into monosaccharides is therefore a complex biochemical process that requires the activities of multiple degradative enzymes with complementary activities tailored to the structural features of a particular substrate. Glucuronoarabinoxylan (GAX) is a major hemicellulose component in softwood, and its structural complexity requires more enzyme specificities to achieve complete hydrolysis compared to glucuronoxylans from hardwood and arabinoxylans from grasses. Results: We report the characterisation of a recombinant α-glucuronidase (Agu115) from Schizophyllum commune capable of removing (4-O-methyl)-glucuronic acid ((Me)GlcA) residues from polymeric and oligomeric xylan. The enzyme is required for the complete deconstruction of spruce glucuronoarabinoxylan (GAX) and acts synergistically with other xylan-degrading enzymes, specifically a xylanase (Xyn10C), an α-l-arabinofuranosidase (AbfA), and a β-xylosidase (XynB). Each enzyme in this mixture showed varying degrees of potentiation by the other activities, likely due to increased physical access to their respective target monosaccharides. The exo-acting Agu115 and AbfA were unable to remove all of their respective target side chain decorations from GAX, but their specific activity was significantly boosted by the addition of the endo-Xyn10C xylanase. We demonstrate that the proposed enzymatic cocktail (Agu115 with AbfA, Xyn10C and XynB) achieved almost complete conversion of GAX to arabinofuranose (Araf), xylopyranose (Xylp), and MeGlcA monosaccharides. Addition of Agu115 to the enzymatic cocktail contributes specifically to 25 % of the conversion. However, traces of residual oligosaccharides resistant to this combination of enzymes were still present after deconstruction, due to steric hindrances to enzyme access to the substrate. Conclusions: Our GH115 α-glucuronidase is capable of finely tailoring the molecular structure of softwood GAX, and contributes to the almost complete saccharification of GAX in synergy with other exo- and endo-xylan-acting enzymes. This has great relevance for the cost-efficient production of biofuels from softwood lignocellulose.Lauren S. McKee, Hampus Sunner, George E. Anasontzis, Guillermo Toriz, Paul Gatenholm, Vincent Bulone, Francisco Vilaplana and Lisbeth Olsso

Approaches in biotechnological applications of natural polymers

Natural polymers, such as gums and mucilage, are biocompatible, cheap, easily available and non-toxic materials of native origin. These polymers are increasingly preferred over synthetic materials for industrial applications due to their intrinsic properties, as well as they are considered alternative sources of raw materials since they present characteristics of sustainability, biodegradability and biosafety. As definition, gums and mucilages are polysaccharides or complex carbohydrates consisting of one or more monosaccharides or their derivatives linked in bewildering variety of linkages and structures. Natural gums are considered polysaccharides naturally occurring in varieties of plant seeds and exudates, tree or shrub exudates, seaweed extracts, fungi, bacteria, and animal sources. Water-soluble gums, also known as hydrocolloids, are considered exudates and are pathological products; therefore, they do not form a part of cell wall. On the other hand, mucilages are part of cell and physiological products. It is important to highlight that gums represent the largest amounts of polymer materials derived from plants. Gums have enormously large and broad applications in both food and non-food industries, being commonly used as thickening, binding, emulsifying, suspending, stabilizing agents and matrices for drug release in pharmaceutical and cosmetic industries. In the food industry, their gelling properties and the ability to mold edible films and coatings are extensively studied. The use of gums depends on the intrinsic properties that they provide, often at costs below those of synthetic polymers. For upgrading the value of gums, they are being processed into various forms, including the most recent nanomaterials, for various biotechnological applications. Thus, the main natural polymers including galactomannans, cellulose, chitin, agar, carrageenan, alginate, cashew gum, pectin and starch, in addition to the current researches about them are reviewed in this article.. }To the Conselho Nacional de Desenvolvimento Cientfíico e Tecnológico (CNPq) for fellowships (LCBBC and MGCC) and the Coordenação de Aperfeiçoamento de Pessoal de Nvíel Superior (CAPES) (PBSA). This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit, the Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462) and COMPETE 2020 (POCI-01-0145-FEDER-006684) (JAT)

Aarhus Regenerative Orthopaedics Symposium (AROS)

The combination of modern interventional and preventive medicine has led to an epidemic of ageing. While this phenomenon is a positive consequence of an improved lifestyle and achievements in a society, the longer life expectancy is often accompanied by decline in quality of life due to musculoskeletal pain and disability. The Aarhus Regenerative Orthopaedics Symposium (AROS) 2015 was motivated by the need to address regenerative challenges in an ageing population by engaging clinicians, basic scientists, and engineers. In this position paper, we review our contemporary understanding of societal, patient-related, and basic science-related challenges in order to provide a reasoned roadmap for the future to deal with this compelling and urgent healthcare problem

Effects of Extractives on the Surface Chemistry and Wettability of High Temperature Chemithermomechanical Pulps

The effect of extractives on surface chemistry and fiber wettability of high temperature chemithermomechanical pulps (HTCTMP) was studied in a model system. Dispersions of abietic acid, oleic acid and a mixture of both were added to acetone extracted HTCTMP made from spruce chips. Surface analysis by electron spectroscopy for chemical analysis, ESCA, showed that adding the dispersions to the extracted pulp decreased the relative oxygen/carbon ratio (O/C). Using scanning electron microscopy, SEM, it was found that the extractives were located mainly on fines and in fiber crossings. A major morphological difference was also found. At the surface of the pulp with only abietic acid added, spherical particles in sizes of 1-20 ?m were observed. These structures were not found on any other sample. Dynamic contact angle (Whilhelmy technique) and water absorption time measurements were used to determine wettability. It was found that adding oleic acid to extracted HTCTMP only slightly affected the wettability of the pulps by increasing the advancing contact angle from 38Zapotitlán to 43Zapotitlán and the water absorption time from 70 to 110 milliseconds. In contrast, adding abietic acid resulted in a tenfold increase in the water absorption time and a contact angle of 56Zapotitlán. The results suggest that the wettability of HTCTMP pulps should be related not only to the amount of extractives but also to the chemical composition of the extractives and to their physical state at the fiber surface

On the interactions between cellulose and xylan, a biomimetic simulation of the hardwood cell wall

The plant cell wall exhibits a hierarchical structure, in which the organization of the constituents on different levels strongly affects the mechanical properties and the performance of the material. In this work, the interactions between cellulose and xylan in a model system consisting of a bacterial cellulose/glucuronoxylan (extracted from aspen, Populus tremula) have been studied and compared to that of a delignified aspen fiber material. The properties of the materials were analyzed using Dynamical Mechanical Analysis (DMA) with moisture scans together with dynamic Infra Red -spectroscopy at dry and humid conditions. The results showed that strong interactions existed between the cellulose and the xylan in the aspen holocellulose. The same kinds of interactions were seen in a water-extracted bacterial cellulose/xylan composite, while unextracted material showed the presence of xylan not interacting with the cellulose. Based on these findings for the model system, it was suggested that there is in hardwood one fraction of xylan that is strongly associated with the cellulose, taking a similar role as glucomannan in softwood.cited By 36</p