Cell Wall Proteomics as a Means to Identify Target Genes to Improve Second‐Generation Biofuel Production

Second‐generation biofuels (B2G) generally uses residues composed of lignocellulosic materials to produce renewable energy (potentially up to 50%), without increasing the planted areas. However, the high cost of enzymes required for cell wall disassembly prior to the saccharification makes the B2G production more expensive yet, compared to the first‐generation biofuels. Designing plants with less lignin, a barrier to B2G production, or facilitating cell wall disassembly by searching for the plant mechanisms can be the way to obtain B2G feasibility. Therewith, plant cell wall proteomics provides valuable information concerning the main cell wall proteins (CWPs) involved in its biosynthesis and rearrangements. Essentially, two plants of the grass family have been studied: sugarcane as a crop amenable to second‐generation ethanol (E2G) production; and Brachypodium distachyon as a model plant amenable to genetic transformation. Cell wall proteomics has allowed the identification of numerous CWPs as well as their fine profiling in different organs and at various developmental stages. Proteins acting on carbohydrates, mostly glycosyl hydrolases, and oxidoreductases, including class III peroxidases and laccases, can be highlighted. Both kinds of CWPs are assumed to contribute to the remodelling of cell wall polysaccharides by enzymatic or non‐enzymatic mechanisms. CWPs present in growing organs could also be attractive candidates since they greatly contribute to cell wall plasticity

Cloning and endogenous expression of a Eucalyptus grandis UDP-glucose dehydrogenase cDNA

UDP-glucose dehydrogenase (UGDH) catalyzes the oxidation of UDP-glucose (UDP-Glc) to UDP-glucuronate (UDP-GlcA), a key sugar nucleotide involved in the biosynthesis of plant cell wall polysaccharides. A full-length cDNA fragment coding for UGDH was cloned from the cambial region of 6-month-old E. grandis saplings by RT-PCR. The 1443-bp-ORF encodes a protein of 480 amino acids with a predicted molecular weight of 53 kDa. The recombinant protein expressed in Escherichia coli catalyzed the conversion of UDP-Glc to UDP-GlcA, confirming that the cloned cDNA encodes UGDH. The deduced amino acid sequence of the cDNA showed a high degree of identity with UGDH from several plant species. The Southern blot assay indicated that more than one copy of UGDH is present in Eucalyptus. These results were also confirmed by the proteomic analysis of the cambial region of 3- and 22-year-old E. grandis trees by 2-DE and LC-MS/MS, showing that at least two isoforms are present. The cloned gene is mainly expressed in roots, stem and bark of 6-month-old saplings, with a lower expression in leaves. High expression levels were also observed in the cambial region of 3- and 22-year-old trees. The results described in this paper provide a further view of the hemicellulose biosynthesis during wood formation in E. grandis