title: Industrial PE-2 strain of Saccharomyces cerevisiae: from alcoholic fermentation to the production of recombinant proteins.
abstract: Saccharomyces cerevisiae is the most important microorganism used in the ethanol fermentation process. The PE-2 strain of this yeast is widely used to produce alcohol in Brazil due to its high fermentation capacity. The aim of the present study was to develop an expression system for recombinant proteins using the industrial PE-2 strain of S. cerevisiae during the alcoholic fermentation process. The protein chosen as a model for this system was CaneCPI-1, a cysteine peptidase inhibitor. A plasmid containing the CaneCPI-1 gene was constructed and yeast cells were transformed with the pYADE4_CaneCPI-1 construct. To evaluate the effect on fermentation ability, the transformed strain was used in the fermentation process with cell recycling. During the nine-hour fermentative cycles the transformed strain did not have its viability and fermentation ability affected. In the last cycle, when the fermentation lasted longer, the protein was expressed probably at the expense of ethanol once the sugars were exhausted. The recombinant protein was expressed in yeast cells, purified and submitted to assays of activity that demonstrated its functionality. Thus, the industrial PE-2 strain of S. cerevisiae can be used as a viable system for protein expression and to produce alcohol simultaneously. The findings of the present study demonstrate the possibility of producing recombinant proteins with biotechnological applications during the ethanol fermentation process.
authors.affiliation: Laboratory of Molecular Biology, Department of Genetics and Evolution, Federal University of São Carlos, Rodovia Washington Luis km 235, 13565-905 São Carlos, SP, Brazil. Electronic address: andreasc@ufscar.br.
authors.forename: Andrea
authors.initials: A
authors.lastname: Soares-Costa
authors.forename: Darlan Gonçalves
authors.initials: DG
authors.lastname: Nakayama
authors.forename: Letícia de Freitas
authors.initials: Lde F
authors.lastname: Andrade
authors.forename: Lucas Ferioli
authors.initials: LF
authors.lastname: Catelli
authors.forename: Ana Paula Guarnieri
authors.initials: AP
authors.lastname: Bassi
authors.forename: Sandra Regina
authors.initials: SR
authors.lastname: Ceccato-Antonini
authors.forename: Flavio
authors.initials: F
authors.lastname: Henrique-Silva
mesh_terms: D015853:Cysteine Proteinase Inhibitors / Q000096:biosynthesis* / Q000235:genetics; D000431:Ethanol / Q000378:metabolism*; D005285:Fermentation; D010940:Plant Proteins / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development*; D031786:Saccharum / Q000235:genetics*
chemical_list: D015853:Cysteine Proteinase Inhibitors; D010940:Plant Proteins; D011994:Recombinant Proteins; D000431:Ethanol