Heterologous production and characterization of a thermostable GH10 family endo-xylanase

GARRIDO, MERCEDES; CAMPOS, ELEONORA; NIDERHAUS, CECILIA; WIRTH, SONIA; INSANI, MARINA

Cordoba

Congreso; LII Reunión Anual Sociedad Argentina de Investigacion en Bioquimica y Biología Molecular; 2016

SAIB

Xylanases show a wide range of biotechnological applications in the feed, pulp and paper industries and production of second generation bioethanol. Development of novel enzymes with enhanced properties for harsh processes requires the bioprospecting and/or engineering of enzymes to be stable and active in acidic or alkaline conditions, high temperatures, oxidative conditions and tolerant to high salinity and/or organic solvents. In this work we present the heterologous expression and characterization of a novel endo-xylanase of glycoside hydrolase family 10 from the white-rot basidiomycete Pycnoporus sanguineus. The coding sequence of mature endo-xylanase GH10ps fused to an N-terminal 6xHis-tag, was cloned in continuous reading frame to the signal sequence of Saccahromyces cerevisiae alfa factor, under the transcriptional control of the methanol-inducible promoter AOX1 in the vector pPIC9, for expression in Pichia pastoris and secretion to the culture medium. Recombinant GH10ps presented thermostable endo-beta-1,4 xylanase activity, with a half-life of 3 hs at 70°C and a stability higher than 48 h at 60°C. Not only was it capable of releasing xylooligosaccahrides and xylose from agricultural waste biomass, but it also acted synergically with comercial cellulases in cellulose saccharification.