title: Proteolytic enzyme engineering: a tool for wool.
abstract: One of the goals of protein engineering is to tailor the structure of enzymes to optimize industrial bioprocesses. In the present work, we present the construction of a novel high molecular weight subtilisin, based on the fusion of the DNA sequences coding for Bacillus subtilis prosubtilisin E and for an elastin-like polymer (ELP). The resulting fusion protein was biologically produced in Escherichia coli , purified and used for wool finishing assays. When compared to the commercial protease Esperase, the recombinant subtilisinE-VPAVG(220) activity was restricted to the cuticle of wool, allowing a significant reduction of pilling, weight loss and tensile strength loss of wool fibers. Here we report, for the first time, the microbial production of a functionalized high molecular weight protease for controlled enzymatic hydrolysis of wool surface. This original process overcomes the unrestrained diffusion and extended fiber damage which are the major obstacles for the use of proteases for wool finishing applications.
authors.affiliation: Department of Biology, Centre of Molecular and Environmental Biology, University of Minho, Braga, Portugal.
authors.forename: Rita
authors.initials: R
authors.lastname: Araújo
authors.forename: Carla
authors.initials: C
authors.lastname: Silva
authors.forename: Raul
authors.initials: R
authors.lastname: Machado
authors.forename: Margarida
authors.initials: M
authors.lastname: Casal
authors.forename: António M
authors.initials: AM
authors.lastname: Cunha
authors.forename: José Carlos
authors.initials: JC
authors.lastname: Rodriguez-Cabello
authors.forename: Artur
authors.initials: A
authors.lastname: Cavaco-Paulo
mesh_terms: D000818:Animals; D002152:Calorimetry, Differential Scanning; D006868:Hydrolysis; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D012697:Serine Endopeptidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D013381:Subtilisins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D013718:Tensile Strength; D014935:Wool
chemical_list: D012697:Serine Endopeptidases; D013381:Subtilisins; C021553:mesentericopeptidase