Detection system for Saccharomyces cerevisiae with phenyl acrylic acid decarboxylase gene (PAD1) and sulphur efflux gene
- PDF / 1,000,648 Bytes
- 5 Pages / 595.276 x 790.866 pts Page_size
- 17 Downloads / 164 Views
ORIGINAL PAPER
Detection system for Saccharomyces cerevisiae with phenyl acrylic acid decarboxylase gene (PAD1) and sulphur efflux gene (SSU1) by multiplex PCR K. M. Archana1 · K. A. Anu‑Appaiah1
Received: 26 February 2017 / Revised: 9 October 2017 / Accepted: 13 October 2017 © Springer-Verlag GmbH Germany 2017
Abstract 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.
Communicated by Erko Stackebrandt. Electronic supplementary material The online version of this article (doi:10.1007/s00203-017-1440-8) contains supplementary material, which is available to authorized users. * K. A. Anu‑Appaiah [email protected] 1
Department of Microbiology and Fermentation Technology, CSIR–Central Food Technological Research Institute, Mysuru, Karnataka, India
Keywords Wine · Saccharomyces cerevisiae · Enological trait · Phenyl acrylic acid decarboxylase gene · Sulphite efflux gene · Multiplex PCR
Introduction Saccharomyces cerevisiae strains have been selected for their enological properties and are used as starters in wine-making (Nardi 2006). Yeast manufacturing companies market a wide variety of dehydrated cultures of various S. cerevisiae strains, and most of the world wine production rely on these commercial starter yeasts. However, they do not enhance the aromatic traits that characterize yeasts isolated from specific geographical areas. Hence, there has been an increased use of native selected yeasts for fermentation (Nardi 2006; Aponte and Blaiotta 2016). The aroma and flavour of wine are derived from grapes. The final quality of wine depends on the ability of microorganisms to produce flavour compounds from flavourless grape-derived precursors. Volatile phenols in wine are formed by the microbial transformation of hydroxyci
Data Loading...
