Paludibacter propionicigenes GH10 xylanase as a tool for enzymatic xylooligosaccharides production from heteroxylans.

Bioconversion of lignocellulosic biomass into value-added products relies on polysaccharides depolymerization by carbohydrate active enzymes. This work reports biochemical characterization of Paludibacter propionicigenes xylanase from GH10 (PpXyn10A) and its application for enzymatic xylooligosaccharides (XOS) production from commercial heteroxylans and liquor of hydrothermally pretreated corn cobs (PCC). PpXyn10A is tolerant to ethanol and NaCl, and releases xylobiose (X2) and xylotriose (X3) as the main hydrolytic products. The conversion rate of complex substrates into short XOS was approximately 30% for glucuronoxylan and 8.8% for rye arabinoxylan, after only 4 h; while for PCC, PpXyn10A greatly increased unbranched XOS yields. B. adolescentis fermentation with XOS from beechwood glucuronoxylan produced mainly acetic and lactic acids. Structural analysis shows that while the glycone region of PpXyn10A active site is well preserved, the aglycone region has aromatic interactions in the +2 subsite that may explain why PpXyn10A does not release xylose.

Recent advances in the enzymatic production and applications of xylooligosaccharides.

The majority of lignocellulosic biomass on the planet originates from plant cell walls, which are complex structures build up mainly by cellulose, hemicellulose and lignin. The largest part of hemicellulose, xylan, is a polymer with a ß-(1→4)-linked xylose residues backbone decorated with α-D-glucopyranosyl uronic acids and/or L-arabinofuranose residues. Xylan is the second most abundant biopolymer in nature, which can be sustainably and efficiently degraded into decorated and undecorated xylooligosaccharides (XOS) using combinations of thermochemical pretreatments and enzymatic hydrolyses, that have broad applications in the food, feed, pharmaceutical and cosmetic industries. Endo-xylanases from different complex carbohydrate-active enzyme (CAZyme) families can be used to cleave the backbone of arabino(glucurono)xylans and xylooligosaccharides and degrade them into short XOS. It has been shown that XOS with a low degree of polymerization have enhanced prebiotic effects conferring health benefits to humans and animals. In this review we describe recent advances in the enzymatic production of XOS from lignocellulosic biomass arabino- and glucuronoxylans and their applications as food and feed additives and health-promoting ingredients. Comparative advantages of xylanases from different CAZy families in XOS production are discussed and potential health benefits of different XOS are presented.

Comparative analysis of two recombinant LPMOs from Aspergillus fumigatus and their effects on sugarcane bagasse saccharification.

Lytic polysaccharide monooxygenases (LPMOs) have been introduced into industrial cocktails used for biomass saccharification due to their capacity to boost enzymatic conversion of recalcitrant cellulose. The genome of the thermotolerant ascomycete Aspergillus fumigatus encodes 7 genes for LPMOs that belong to auxiliary activity family 9 (AA9). Here, we cloned, successfully expressed and performed biochemical evaluation of two CBM-less A. fumigatus LPMOs (AfAA9A and AfAA9B). A high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) analysis demonstrated that AfAA9A and AfAA9B are able to oxide cellulose at C1 and C1/C4 positions, respectively. Synergic effects of LPMOs (separately and in combination) with cellulases were investigated. Supplementation of Celluclast 1.5 L with a low concentration of AfAA9B improved in 20 % the saccharification of sugarcane bagasse pretreated by steam explosion (SEB), while AfAA9A did not improvethe saccharification. Analysis of the hydrolyzed biomass by confocal laser scanning microscopy (CLSM) showed the LPMOs are promoting lignin oxidation in the lignocellulosic material. This study complements the available results concerning the utilization of LPMOs in the enzymatic saccharification of lignocellulosic biomass.

Production of prebiotic xylooligosaccharides from arabino- and glucuronoxylan using a two-domain Jonesia denitrificans xylanase from GH10 family.

Development of a more environmentally sustainable society is based on the maximum use of renewable carbon sources and their valorization of environmentally-friendly green technologies. This includes a thorough use of plant biomass and agricultural residues for the production of value-added bioproducts. Xylan is the second most abundant biopolymer in nature which can be sustainable converted into pentoses and xylooligosaccharides, that have wide applications in the food, feed, pharmaceutical, and cosmetic industry. Within the scope of present study, we biochemically characterized two-domain GH10 xylanase from Jonesia denitrificans (JdXyn10A) and evaluated its applicability for production of xylooligosaccharides (XOS). JdXyn10A has a specific activity of 84 ± 2 U/mg and 65 ± 5 U/mg when acting on beechwood glucuronoxylan and rye arabinoxylan, respectively. The enzyme is stable in a wide pH range and is tolerant to high concentrations of NaCl and ethanol. Interestingly, the profile of products released by the enzyme is predominant in xylobiose and xylotriose, with a very low fraction of xylose which is desirable for XOS production. The efficiencies of enzymatic conversion of beechwood glucuronoxylan and rye arabinoxylan are 47.67 % and 26.01 %, respectively, after 6 h of enzymatic hydrolysis only. Structural comparison between the JdXyn10A homology model and the structure from its homologous that while the glycone region of its active site is well preserved, the aglycone region presents structural differences in the +2 subsite that may explain why JdXyn10A does not release xylose.

Magnetoreception in multicellular magnetotactic prokaryotes: a new analysis of escape motility trajectories in different magnetic fields.

Magnetotactic microorganisms can be found as unicellular prokaryotes, as cocci, vibrions, spirilla and rods, and as multicellular organisms. Multicellular magnetotactic prokaryotes are magnetotactic microorganisms composed by several magnetotactic bacteria organized almost in a spherical helix, and one of the most studied is Candidatus Magnetoglobus multicellularis. Several studies have shown that Ca. M. multicellularis displays forms of behavior not well explained by magnetotaxis. One of these is escape motility, also known as "ping-pong" motion. Studies done in the past associated the "ping-pong" motion to some magnetoreceptive behavior, but those studies were never replicated. In the present manuscript a characterization of escape motility trajectories of Ca. M. multicellularis was done for several magnetic fields, considering that this microorganism swims in cylindrical helical trajectories. It was observed that the escape motility can be separated into three phases: (I) when the microorganism jumps from the drop border, (II) where the microorganism moves almost perpendicular to the magnetic field and (III) when the microorganism returns to the drop border. The total time of the whole escape motility, the time spent in phase II and the displacement distance in phase I decreases when the magnetic field increases. Our results show that the escape motility has several characteristics that depend on the magnetic field and cannot be understood by magnetotaxis, with a magnetoreceptive mechanism being the best explanation.

Transformation of xylan into value-added biocommodities using Thermobacillus composti GH10 xylanase.

Enzymatic transformation of xylans into renewable fuels and value-added products is mediated by xylanases. Here we describe the biochemical and X-ray structural characterization of Thermobacillus composti GH10 xylanase (TcXyn10A) at 2.1 Å resolution aiming to unravel details of its recognition of glucurono- and arabinoxylan at a molecular level. TcXyn10A improves the efficiency of pretreated lignocellulosic biomass hydrolysis by a commercial enzyme cocktail causing a 15.35 % increase in xylose release and 4.38 % glucose release after 24 h of reaction. The enzyme releases predominantly xylobiose and xylotriose, as well as MeGlcA3 × 3 (from beechwood glucuronoxylan) and a range of decorated xylooligosaccharides (XOS) from rye arabinoxylan, with Ara2 × 2 being the major product. The enzyme liberates XOS with the yields of 29.09 % for beechwood glucuronoxylan and 16.98 % for rye arabinoxylan. Finally, TcXyn10A has a high thermal stability, halotolerance, and resistance to ethanol, biochemical properties that can be desirable for a number of industrial applications.