title: Recombinant expression of a GH12 β-glucanase carrying its own signal peptide from Stachybotrys atra in yeast and filamentous fungi.
abstract: The β-glucanase Cel12A gene from Stachybotrys atra has been cloned and heterologously expressed in Aspergillus nidulans and Saccharomyces cerevisiae. The recombinant strains constructed, contained the exonic sequence of cel12A including its own signal peptide coding sequence. SDS-PAGE and zymography revealed that recombinant Cel12A has a molecular mass of 24 kDa which agrees with that deduced from its amino acid sequence, indicating that it is expressed in the non-glycosylated active form. Recombinant A. nidulans showed about eightfold greater activity yield than S. cerevisiae recombinant strain, namely 0.71 and 0.09 β-glucanase Units/ml of culture, respectively. In both host strains most of the activity was secreted to the extracellular media, evidencing the functionality of Cel12A signal peptide in yeast and fungi. This novel signal peptide might facilitate the expression and efficient secretion of other recombinant proteins difficult to secrete.
authors.affiliation: Department of Microbiology, Faculty of Biology, University of Barcelona, Avinguda Diagonal 643, 08028, Barcelona, Spain.|Institute of Biotechnology RWTH, Aachen University, Worringerweg 3, 52074, Aachen, Germany.
authors.forename: Pere
authors.initials: P
authors.lastname: Picart
authors.affiliation: Instituto de Agroquímica y Tecnología de Alimentos, Consejo Superior de Investigaciones Científicas (IATA-CSIC), Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain.
authors.forename: Margarita
authors.initials: M
authors.lastname: Orejas
authors.affiliation: Department of Microbiology, Faculty of Biology, University of Barcelona, Avinguda Diagonal 643, 08028, Barcelona, Spain. fpastor@ub.edu.
authors.forename: F I Javier
authors.initials: FI
authors.lastname: Pastor
mesh_terms: D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000235:genetics; D003001:Cloning, Molecular; D005818:Genetic Engineering; D006026:Glycoside Hydrolases / Q000235:genetics* / Q000378:metabolism*; D008970:Molecular Weight; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013191:Stachybotrys / Q000201:enzymology* / Q000235:genetics
chemical_list: D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D006026:Glycoside Hydrolases
keywords: Aspergillus nidulans; GH12; Saccharomyces cerevisiae; Signal peptide; Stachybotrys; β-Glucanase