AtFUT4 and AtFUT6 are arabinofuranose-specific fucosyltransferases

The bulk of plant biomass is comprised of plant cell walls, which are complex polymeric networks, composed of diverse polysaccharides, proteins, polyphenolics, and hydroxyproline-rich glycoproteins (HRGPs). Glycosyltransferases (GTs) work together to synthesize the saccharide components of the plant cell wall. The Arabidopsis thaliana fucosyltransferases (FUTs), AtFUT4, and AtFUT6, are members of the plant-specific GT family 37 (GT37). AtFUT4 and AtFUT6 transfer fucose (Fuc) onto arabinose (Ara) residues of arabinogalactan (AG) proteins (AGPs) and have been postulated to be non-redundant AGP-specific FUTs. AtFUT4 and AtFUT6 were recombinantly expressed in mammalian HEK293 cells and purified for biochemical analysis. We report an updated understanding on the specificities of AtFUT4 and AtFUT6 that are involved in the synthesis of wall localized AGPs. Our findings suggest that they are selective enzymes that can utilize various arabinogalactan (AG)-like and non-AG-like oligosaccharide acceptors, and only require a free, terminal arabinofuranose. We also report with GUS promoter-reporter gene studies that AtFUT4 and AtFUT6 gene expression is sub-localized in different parts of developing A. thaliana roots

Optimization of xylanase production by filamentous fungi in solid state fermentation and scale-up to horizontal tube bioreactor

Five microorganisms, namely Aspergillus niger CECT 2700, A. niger CECT 2915, A. niger CECT 2088, Aspergillus terreus CECT 2808, and Rhizopus stolonifer CECT 2344, were grown on corncob to produce cell wall polysaccharide-degrading enzymes, mainly xylanases, by solid-state fermentation (SSF). A. niger CECT 2700 produced the highest amount of xylanases of 504±7 U/g dry corncob (dcc) after 3 days of fermentation. The optimization of the culture broth (5.0 g/L NaNO3, 1.3 g/L (NH4)2SO4, 4.5 g/L KH2PO4, and 3 g/L yeast extract) and operational conditions (5 g of bed loading, using an initial substrate to moistening medium of 1:3.6 (w/v)) allowed increasing the predicted maximal xylanase activity up to 2,452.7 U/g dcc. However, different pretreatments of materials, including destarching, autoclaving, microwave, and alkaline treatments, were detrimental. Finally, the process was successfully established in a laboratory-scale horizontal tube biore- actor, achieving the highest xylanase activity (2,926 U/g dcc) at a flow rate of 0.2 L/min. The result showed an overall 5.8-fold increase in xylanase activity after optimization of culture media, operational conditions, and scale-up.We are grateful to the Spanish Ministry of Science and Innovation for the financial support of this work (project CTQ2011-28967), which has partial financial support from the FEDER funds of the European Union; to the Leonardo da Vinci Programme for founding the stay of Felisbela Oliveira in Vigo University; to MAEC-AECID (Spanish Government) for the financial support for Perez-Bibbins, B. and to Spanish Ministry of Education, Culture and Sports for Perez-Rodriguez's FPU; and to Solla E. and Mendez J. (CACTI-University of Vigo) for their excellent technical assistance in microscopy

Pretreatment and enzymatic hydrolysis of lignocellulosic biomass for reducing sugar production

Conversion of lignocellulosic biomass into reducing sugar has contributed to an alternative use of lignocellulose source, especially in the production of value-added products such as amino acids, biofuels, and vitamins. In the bioconversion process, pretreatment of lignocellulosic biomass is important to enhance the accessibility of enzyme hydrolysis, thus increasing the yield of reducing sugar. Lignocellulosic biomass has a very complex arrangement of structure that needs a proper study in pretreatment and enzymatic hydrolysis process to obtain an optimum yield of reducing sugar. This chapter discusses chemical and enzymatic pretreatment methods that are commonly applied to effectively modify the chemical structures of lignocellulosic biomass. Acid pretreatment using dilute sulfuric acid (H2SO4) is the most commonly employed for chemical pretreatment while sodium hydroxide (NaOH) is the most commonly applied for alkaline pretreatment because of its ability to delignify biomass. Then, enzymatic hydrolysis of lignocellulosic biomass for the production of reducing sugar is discussed in detail. The kinetics and optimization of hydrolysis which are the key parameters that determine the yields of reducing sugar are also presented. The right pretreatment method combined with an efficient hydrolysis process will ensure successful conversion of lignocellulosic biomass into reducing sugar, thus providing a sustainable production of reducing sugar from biomass for various applications