Identification of the structural gene encoding maltase within the MAL6 locus of Saccharomyces carlsbergensis

Abstract

Saccharomyces yeast strains able to ferment maltose carry at least one member of a family of MAL loci: MAL1, MAL2, MAL3, MAL4, and MAL6. The MAL6 locus has been cloned and shown to be a cluster of at least three transcribed regions, all of which are required for maltose fermentation. Transcription at two of these genes, MAL61 and MAL62, is both induced by maltose and repressed by glucose. The third gene, MAL63, appears to encode a regulatory product controlling maltose fermentation. In this report, we demonstrate that the MAL62 gene is the structural gene coding for the enzyme maltase. Strain 332-5A is a maltose fermenter of the genotype MAL6 mal1(0). Integrative disruption of the MAL62 gene of the MAL6 locus produces a strain which is still capable of fermenting maltose, but which synthesizes a more heat-labile form of maltase than the undisrupted strain. Synthesis of this more heat-labile maltase was shown to be linked to the mal1(0) locus present in the strain. Integrative disruption of both the MAL62 gene and the MAL62-homologous sequence present at the mal1(0) locus produces a nonfermenter which is unable to synthesize maltase. These results identify MAL62 as the maltase structural gene.