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Overexpression of the glucoamylase-encoding STA1 gene of Saccharomyces cerevisiae var. diastaticus in laboratory and industrial strains of Saccharomyces
Authors:Lorena Latorre-García  Ana Cristina Adam and Julio Polaina
Institution:(1) Instituto de Agroquímica y Tecnología de Alimentos, CSIC, Polígono de la Coma s/n, Paterna, Valencia, 46980, Spain;(2) Present address: Instituto de Biología Molecular y Celular de Plantas (C.S.I.C.–U.P.V.), Campus de la Universidad Politécnica de Valencia, Avda. de los Naranjos s/n, Valencia, 46022, Spain;(3) Instituto de Agroquímica y Tecnología de Alimentos, Consejo Superior de Investigaciones Científicas, Apdo. de Corrreos 73, Burjassot, Valencia, 46100, Spain
Abstract:Production of glucoamylase encoded by the Saccharomyces cerevisiae (var. diastaticus) STA1 gene has been assayed in laboratory S. cerevisiae strains of different ploidy and in different industrial Saccharomyces strains, in which STA1 was expressed under control of an inducible promoter. Highest enzyme activity was achieved with a tetraploid strain constructed by crossing preselected parental strains. Maximal glucoamylase production correlated with heterogeneity in enzyme mass, likely due to incomplete glycosylation, suggesting that the secretion-glycosylation process is the limiting step in the production of the STA-encoded glucoamylase by Saccharomyces. Industrial strains showed quite different capacity to produce glucoamylase. High production was achieved with a S. pastorianus brewer’s strain. Overall, our results allowed the selection of strains capable of yielding a high level of glucoamylase and suggest specific approaches for further enhancing this capability.
Keywords:Ethanol  Fermentation  Industrial strains  Starch  Yeasts
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