Detection system for Saccharomyces cerevisiae with phenyl acrylic acid decarboxylase gene (PAD1) and sulphur efflux gene (SSU1) by multiplex PCR.

Production of wine with selected yeast strains is a common enological practice followed for the production of wine with desirable organoleptic properties and to guarantee the homogeneity of successive vintages. Sulphur dioxide tolerance and phenyl acrylic acid resistance are the enological traits essential for the survivability of the yeast during fermentation. The present study describes the detection of S. cerevisiae with enological traits, such as phenyl acrylic acid resistance and sulphur dioxide tolerance in a single test. Phenyl acrylic acid decarboxylase (PAD1) and sulphite efflux genes (SSU1) were detected by multiplex PCR, thus confirming the specificity of primers. A single cocktail of all reagents required for the simultaneous detection of both these genes was designed. The ready-to-use formulation optimized was stable at 4 and - 20 °C for 6 months. The amplification of phenyl acrylic acid decarboxylase and sulphite efflux genes, validated the suitability of the ready-to-use formulation for the detection of S. cerevisiae in food samples. The ready-to-use formulation optimized, minimizes the end user requirements for the detection of S. cerevisiae. Thus, the method was suitable for the identification of S. cerevisiae strains from a mixture of yeast prior to the sequencing analysis, thereby reducing the cost and time of screening.