title: Characterization of the cellulolytic secretome of Trichoderma harzianum during growth on sugarcane bagasse and analysis of the activity boosting effects of swollenin.
abstract: This study demonstrates the production of an active enzyme cocktail produced by growing Trichoderma harzianum on sugarcane bagasse. The component enzymes were identified by LCMS-MS. Glycosyl hydrolases were the most abundant class of proteins, representing 67% of total secreted protein. Other carbohydrate active enzymes involved in cell wall deconstruction included lytic polysaccharide mono-oxygenases (AA9), carbohydrate-binding modules, carbohydrate esterases and swollenin, all present at levels of 1%. In total, proteases and lipases represented 5 and 1% of the total secretome, respectively, with the rest of the secretome being made up of proteins of unknown or putative function. This enzyme cocktail was efficient in catalysing the hydrolysis of sugarcane bagasse cellulolignin to fermentable sugars for potential use in ethanol production. Apart from mapping the secretome of T. harzianum, which is a very important tool to understand the catalytic performance of enzyme cocktails, the gene coding for T. harzianum swollenin was expressed in Aspergillus niger. This novel aspect in this work, allowed increasing the swollenin concentration by 95 fold. This is the first report about the heterologous expression of swollenin from T. harzianum, and the findings are of interest in enriching enzyme cocktail with this important accessory protein which takes part in the cellulose amorphogenesis. Despite lacking detectable glycoside activity, the addition of swollenin of T. harzianum increased by two-fold the hydrolysis efficiency of a commercial cellulase cocktail. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:327-336, 2016.
authors.affiliation: LADEBIO, Centre of Biofuels, Oil and its Derivatives, School of Chemistry, Department of Biochemical Engineering, Centre of Technology, Federal University of Rio De Janeiro, Rio De Janeiro, 21941-909, Brazil.
authors.forename: Vanessa
authors.initials: V
authors.lastname: A L Rocha
authors.affiliation: LADEBIO, Centre of Biofuels, Oil and its Derivatives, School of Chemistry, Department of Biochemical Engineering, Centre of Technology, Federal University of Rio De Janeiro, Rio De Janeiro, 21941-909, Brazil.
authors.forename: Roberto
authors.initials: R
authors.lastname: N Maeda
authors.affiliation: LADEBIO, Centre of Biofuels, Oil and its Derivatives, School of Chemistry, Department of Biochemical Engineering, Centre of Technology, Federal University of Rio De Janeiro, Rio De Janeiro, 21941-909, Brazil.
authors.forename: Nei
authors.initials: N
authors.lastname: Pereira
authors.affiliation: MasonCNAP, Dept. of Biology, University of York, Wentworth Way, York, YO10 5DD, U.K.
authors.forename: Marcelo
authors.initials: M
authors.lastname: F Kern
authors.affiliation: MasonCNAP, Dept. of Biology, University of York, Wentworth Way, York, YO10 5DD, U.K.
authors.forename: Luisa
authors.initials: L
authors.lastname: Elias
authors.affiliation: MasonCNAP, Dept. of Biology, University of York, Wentworth Way, York, YO10 5DD, U.K.
authors.forename: Rachael
authors.initials: R
authors.lastname: Simister
authors.affiliation: MasonCNAP, Dept. of Biology, University of York, Wentworth Way, York, YO10 5DD, U.K.
authors.forename: Clare
authors.initials: C
authors.lastname: Steele-King
authors.affiliation: MasonCNAP, Dept. of Biology, University of York, Wentworth Way, York, YO10 5DD, U.K.
authors.forename: Leonardo D
authors.initials: LD
authors.lastname: Gómez
authors.affiliation: MasonCNAP, Dept. of Biology, University of York, Wentworth Way, York, YO10 5DD, U.K.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: McQueen-Mason
mesh_terms: D055162:Biocatalysis; D044602:Cellulases / Q000032:analysis* / Q000378:metabolism; D002482:Cellulose / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D020543:Proteome / Q000737:chemistry / Q000378:metabolism*; D031786:Saccharum / Q000737:chemistry / Q000378:metabolism*; D014242:Trichoderma / Q000737:chemistry / Q000378:metabolism*
chemical_list: D020543:Proteome; D002482:Cellulose; C027433:bagasse; D044602:Cellulases
keywords: Trichoderma harzianum; enzymatic hydrolysis; secretome; sugarcane bagasse; swollenin