Protein engineering and covalent modification of trichoderma reesei cellulases in pichia pastoris for textile biofinishing

摘要: Cellulase enzymes have been extensively used for the biopolishing of cellulosic fabrics but they are inefficient to prevent pilling in viscose fabrics. Moreover, their application causes a loss fabric strength due aggressive action enzymes. One solution this problem is design and production with increased molecular weights so that cellulases would be limited surface. In framework study, cellulase formulations can ameliorate tensile were designed produced . For purpose, both protein engineering chemical modification methods seperately combination obtain desired properties. Trichoderma reesei Endoglucanase I (EGI), III (EGIII), Cellobiohydrolase (CBHI) successfully cloned expressed Pichia pastoris under control AOX1 promoter mg/L quantities. A loop mutant EGI, (EGI_L5) was prepared by introduction ten aminoacid long modelling site directed mutagenesis creation hotspots crosslinking enzyme. The enzyme crosslinked using aggregate (CLEA) technology. effect codon optimization on EGI analyzed. inserting second catalytic domain thereby forming bicatalytic (EGI_BC) weight. All recombinant laboratory scale fermenter characterized. commercial preparation CLEA technology fractionated according particle size. effects native, engineered chemically modified evaluated. It found technology, EGI_L5 P. improved values 20 % without much