title: Functional diversity for biomass deconstruction in family 5 subfamily 5 (GH5_5) of fungal endo-β1,4-glucanases.
abstract: Endo-β1,4-glucanases in glycosyl hydrolase family 5 (GH5) are ubiquitous enzymes in the multicellular fungi and are common components of enzyme cocktails for biomass conversion. We recently showed that an endo-glucanase of subfamily 5 of GH5 (GH5_5) from Sporotrichum thermophile (StCel5A) was more effective at releasing glucose from pretreated corn stover, when part of an eight-component synthetic enzyme mixture, compared to its closely related counterpart from Trichoderma reesei, TrCel5A. StCel5A and TrCel5A belong to different clades of GH5_5 (GH5_5_1 and GH5_5_2, respectively). To test whether the superior activity of StCel5A was a general property of all enzymes in the GH5_5_2 clade, StCel5A, TrCel5A, and two additional members of each subfamily were expressed in a common host that had been engineered to suppress its native cellulases (T. reesei Δxyr1) and compared against each other alone on pure substrates, in synthetic mixtures on pure substrates, and against each other in synthetic mixtures on real biomass. The results indicated that superiority is a unique property of StCel5A and not of GH5_5_2 generally. The six Cel5A enzymes had significant differences in relative activities on different substrates, in specific activities, and in sensitivities to mannan inhibition. Importantly, the behavior of the six endo-glucanases on pure cellulose substrates did not predict their behavior in combination with other cellulolytic enzymes on a real lignocellulosic biomass substrate.
authors.affiliation: Department of Energy Plant Research Laboratory, Department of Plant Biology, and DOE Great Lakes Bioenergy Research Center, Michigan State University, E. Lansing, MI, 48824, USA.
authors.forename: Bingyao
authors.initials: B
authors.lastname: Li
authors.affiliation: Department of Energy Plant Research Laboratory, Department of Plant Biology, and DOE Great Lakes Bioenergy Research Center, Michigan State University, E. Lansing, MI, 48824, USA. walton@msu.edu.
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Walton
mesh_terms: D018533:Biomass; D001709:Biotechnology / Q000379:methods; D002480:Cellulase / Q000737:chemistry* / Q000145:classification / Q000378:metabolism*; D002482:Cellulose; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006868:Hydrolysis; D013175:Sporothrix / Q000201:enzymology / Q000378:metabolism*; D014242:Trichoderma / Q000201:enzymology / Q000378:metabolism*; D003313:Zea mays
chemical_list: D005656:Fungal Proteins; D002482:Cellulose; D002480:Cellulase; D005947:Glucose
keywords: Biomass; Cellulase; Corn stover; Endo-β1,4-glucanase; Lytic polysaccharide mono-oxygenase; Sporotrichum; Trichoderma; cdna1; xyr1