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 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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Aponte M, Blaiotta G (2016) Selection of an autochthonous Saccharomyces cerevisiae strain for the vinification of “Moscato di Saracena”, a Southern Italy (Calabria region) passito wine. Food Microbiol 54:30–39
Archana KM, Ramasamy R, Anu-Appaiah KA (2015) Correlation between ethanol stress and cellular fatty acid composition of alcohol producing non-Saccharomyces in comparison with Saccharomyces cerevisiae by multivariate techniques. J Food Sci Technol 52:6770–6776
Clausen M, Lamb CJ, Megnet R, Doerner PW (1994) PAD1 encodes phenylacrylic acid decarboxylase which confers resistance to cinnamic acid in Saccharomyces cerevisiae. Gene 142:107–112
Cordero-Bueso G, Rodriguez ME, Garriado C, Cantoral JM (2017) Rapid and non culture-dependent assay based on multiplex PCR-SSR analysis for monitoring inoculated yeast strains in industrial wine fermentations. Arch Microbiol 199(1):135–143
Depak MB, Anu-Appaiah KA (2015) Fatty acid profiling of isolated toddy yeasts and their molecular identification. Int J Curr Res 7(4):15189–15194
Der-Shyan S, Yun-Ting W, Chia-Yin L (2000) Rapid detection of polyhydroxyalkanoate accumulating bacteria isolated from the environment by colony. PCR Microbiol 146:2019–2025
Fujihara H, Hino M, Takashita H, Kajwara Y, Okamoto K, Furukawa K (2014) Efficient screening of environmental isolates for Saccharomyces cerevisiae strains that are suitable for brewing. Biosci Biotechnol Biochem 78(6):1086–1089
Fujita SI, Senda Y, Nakaguchi S, Hashimoto T (2001) Multiplex PCR using internal transcribed spacer 1 and 2 regions for rapid detection and identification of yeast strains. J Clin Microbiol 39:3617–3622
Hanna M, Xiao W (2006) Isolation of nucleic acids. In: In: Yeast protocols, 2nd edn. Human Press Inc, Totowa, pp 15–20
Hoffman CS, Winston F (1987) A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57:267–272
Ivey ML, Phister TG (2011) Dection and identification of mioroorganisms in wine: a review of molecular techniques. J Ind Microbiol Biotechnol 38:1619–1634
Jeyaram K, Singh WM, Capece A, Romano P (2008) Molecular identification of yeast species associated with “Hamei”—a traditional starter used for rice wine production in Manipur, India. Int J Food Microbiol 124:115–125
Kurtzman CP, Robnett CJ (1997) Identification of clinically important ascomycetous yeasts based on nucleotide divergence in the 5′ end of the large-subunit (26S) ribosomal DNA gene. J Clin Microbiol 35:1216–1223
Nardi T (2006) Molecular approaches for the individuation and characterization of technological and quality traits Approcci molecolari. Thesis. Universita’Degli Studi di Padova
Nardi T, Corich V, Giacomini A, Blondin B (2010) A sulphite-inducible form of the sulphite efflux gene SSU1 in a Saccharomyces cerevisiae wine yeast. Microbiology 156:1686–1696
Nelson L (2008) The production of volatile phenols by wine microorganisms. Thesis. Stellenobosch University
Park H, Bakalinsky AT (2000) SSU1 mediates sulphite efflux in Saccharomyces cerevisiae. Yeast 16:881–888
Shinohara T, Kubodera S, Yanagida F (2000) Distribution of phenolic yeasts and production of phenolic off-flavors in wine fermentation. J Biosci Bioeng 90(1):90–97
Somashekar KL, Anu Appaiah KA (2013) Coffee cherry husk—a potential feed stock for alcohol production. Int J Environ Waste Manag 11(4):410–419
Vaudano E, Garcia-Moruno E (2008) Discrimination of Saccharomyces cerevisiae wine strains using microsatellite multiplex PCR and band pattern analysis. Food Microbiol 25:56–64
Acknowledgements
The authors thank Director, CSIR-Central Food Technological Research Institute (CFTRI) for providing access to the resources necessary for the completion of this study. The first author acknowledges the Fellowship by Department of Science and Technology (DST), Government of India under INSPIRE fellowship program.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by Erko Stackebrandt.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Archana, K.M., Anu-Appaiah, K.A. Detection system for Saccharomyces cerevisiae with phenyl acrylic acid decarboxylase gene (PAD1) and sulphur efflux gene (SSU1) by multiplex PCR. Arch Microbiol 200, 275–279 (2018). https://doi.org/10.1007/s00203-017-1440-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00203-017-1440-8