Induced expression of bacterial beta-glucosidase activity in Saccharomyces.
The bglA gene which encodes a beta-glucosidase from Bacillus polymyxa, has been expressed in Saccharomyces cerevisiae under control of the yeast CYC-GAL promoter. Strains have been constructed which carry the gene in different locations: in a multicopy plasmid, a single integration at the URA3 locus, or multiple integrations at the RDN1 locus. Integrative transformation at RDN1 yielded genetically stable clones with a high level of beta-glucosidase activity. Coordinated overexpression of the GAL4 inducer protein further increased the level of enzyme activity, although eventually caused the lysis of the cultures. Diploid, triploid and tetraploid strains derived from the transformants with multiple integrations were constructed and expression of beta-glucosidase activity in different conditions of growth was assayed. While per-cell activity increased with ploidy, specific activity was about the same in strains of equivalent genotype regardless of ploidy. Genetically stable and regulated expression in Saccharomyces of beta-glucosidase activity is interesting for the development of strains able to ferment beta-glycosidic sugars (i.e. cellobiose and lactose). From another point of view, the bglA product proved to be a convenient reporter enzyme to monitor heterologous gene expression.
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Instituto de Agroquímica y Tecnología de Alimentos, Consejo Superior de Investigaciones Cientificas, Valencia, Spain.
Rubio-Texeira
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Polaina
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J
D001407:Bacillus / Q000201:enzymology*; D005285:Fermentation; D016681:Genome, Fungal; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic; D001617:beta-Glucosidase / Q000096:biosynthesis* / Q000235:genetics
D001617:beta-Glucosidase