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 readyto-
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.