Simultaneous production of xylooligosaccharides and antioxidant compounds from sugarcane bagasse via enzymatic hydrolysis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Advances in industrial biotechnology offer potential opportunities for economic utilization of agroindustrial residues such as sugarcane bagasse, which is the major by-product of the sugarcane industry. Due to its abundant availability and despite the complex chemical composition, it can be considered an ideal substrate for microbial processes for the production of value-added products. In the present study we evaluated the enzymatic production of xylooligosaccharides (XOS) and antioxidant compounds from sugarcane bagasse using XynZ from Clostridium thermocellum, a naturally chimeric enzyme comprising activities of xylanase and feruloyl esterase along with a carbohydrate binding module (CBM6). In order to reveal the biotechnological potential of XynZ, the XOS released after enzymatic hydrolysis using different substrates were characterized by capillary electrophoresis and quantified by high performance anion exchange chromatography. In parallel, the antioxidant capacity related to the release of phenolic compounds was also determined. The results indicated noteworthy differences regarding the amount of XOS and antioxidant phenolic compounds produced, as well as the XOS profile, functions of the pre-treatment method employed. The ability of XynZ to simultaneously produce xylooligosaccharides, natural probiotics, phenolic compounds and antioxidant molecules from natural substrates such as sugarcane bagasse demonstrated the biotechnological potential of this enzyme. Production of value-added products from agro-industrial residues is of great interest not only for advancement in the biofuel field, but also for pharmaceutical and food industries. (C) 2013 Elsevier B.V. All rights reserved.52770775Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)CNPq [474022/2011-4, 310177/2011-1, 142685/2010-0]FAPESP [2008/58037-9, 2011/14200-6, 2012/18859-5, 2013/03061-0

Biomass-to-bio-products application of feruloyl esterase from Aspergillus clavatus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The structural polysaccharides contained in plant cell walls have been pointed to as a promising renewable alternative to petroleum and natural gas. Ferulic acid is a ubiquitous component of plant polysaccharides, which is found in either monomeric or dimeric forms and is covalently linked to arabinosyl residues. Ferulic acid has several commercial applications in food and pharmaceutical industries. The study herein introduces a novel feruloyl esterase from Aspergillus clavatus (AcFAE). Along with a comprehensive functional and biophysical characterization, the low-resolution structure of this enzyme was also determined by small-angle X-ray scattering. In addition, we described the production of phenolic compounds with antioxidant capacity from wheat arabinoxylan and sugarcane bagasse using AcFAE. The ability to specifically cleave ester linkages in hemicellulose is useful in several biotechnological applications, including improved accessibility to lignocellulosic enzymes for biofuel production.971567596767Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FAPESP [2008/58037-9, 2011/02169-7, 2009/17956-4]CNPq [475022/2011-4, 310177/2011-1

MOESM1 of Lignolytic-consortium omics analyses reveal novel genomes and pathways involved in lignin modification and valorization

Additional file 1: Table S1. Compounds identified by gas chromatography–mass spectrometry (GC-MS) in the lignin-waste stream used for establishment of the lignin-degrading microbial community (LigMet). Table S2. Sequencing statistics and data processing of amplicon libraries constructed for profiling LigMet and soil samples analyzed. Table S3. Diversity and richness indices of the LigMet and soil samples based on 16S rRNA and ITS2 region sequences. Table S4. Protozoa identified in LigMet based on 18S rRNA sequencing. Table S5. Assembly statistics from draft genomes recovered from LigMet (all assemblies). Table S6. Genome statistics of Paenarthrobacter sp. str. HW13. Figure S1. Microbial growth was monitored by OD 600 nm, observing exponential growing during the first 40 hours of consortium growth. The consumption of reducing sugars over time as monitored by DNS, the exponential phase was completed after the first 40 hours of growth when monitoring sugar consumption. Figure S2. Rarefaction curves based on targeted sequencing of 16S rRNA gene amplicons derived from the LigMet (A) and sugarcane soil (B) samples. The rarefaction curves of each biological replicate are shown in different colors. Figure S3. Rarefaction curves based on targeted sequencing of the ITS2 region derived from the LigMet sample. The rarefaction curves of each biological replicate are shown in different colors. Figure S4. The taxonomic profiles from LigMet and sugarcane soil samples at the class level based on 16S rRNA gene amplicon. The respective relative abundances of each biological replicate for LigMet and sugarcane soil are shown. Figure S5. The archaeal phylum abundance in LigMet and sugarcane soil sample. The relative abundance is shown in percentage for each biological replicate of the LigMet and sugacarcane soil. Figure S6. Metabolic pathways related to aromatic compound degradation identified in LigMet according to KEGG automatic annotation. Figure S7. Classification of the predicted proteins from the LigMet according to the dbCAN database. Figure S8. Predicted auxiliary activity (AA) and carbohydrate esterase (CE) families from LigMet and draft genomes, based on the dbCAN database. AA and CE families are related to peroxidase activity and break down of lignin ester cross links, respectively. Figure S9. Phylogenetic relationship of the strain HW13 relative to the most closely related strains of the genus Paenarthrobacter. EzBioCloud webserver was used to perform a similarity-based search of HW13 16S rRNA to retrieve the most closely related sequences. The resulting 16S rRNA sequences were aligned using the MAFFT v7.299b software. A phylogenetic tree was inferred using the maximum likelihood method implemented in RAxML v8.2.0, evolutionary distances were based on the GTRGAMMAI model, inferred as the best model by jModelTest2. Numbers at the nodes are percentages of bootstrap values obtained by repeating the analysis 1,000 times. The type strains are marked with a superscript ‘T’. Accession numbers are shown in parentheses. Figure S10. Phylogenetic relationships among feruloyl-CoA synthetase (upper) and Enoyl-CoA hydratase/aldolase. The phylogenetic tree was generated using amino acid sequences retrieved from NCBI and Uniprot database. The sequences were aligned using MAFFT v7.299b software [5]. The phylogenetic tree was reconstructed using maximum likelihood method implemented in RAxML v8.2.0 [6], evolutionary distances were based on the GTRGAMMAI model, inferred as the best model by jModelTest2 [7]. The bootstrap values (1,000 replicate runs, shown as %) higher than 70 % are represented. Accession numbers are listed in parentheses. The FerA_B3 and FerB_B11 amino acid sequence retrieved from LigMet data set is printed in bold