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Efficient recovery of whole cell proteins in Oenococcus oeni—a comparison of different extraction protocols for high-throughput malolactic starter applications

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Abstract

In this study, we compared different total protein extraction protocols to achieve highly efficient isolation and purification of total proteins for the specific protein profiling of Oenococcus oeni. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns obtained for the different extraction protocols revealed not only a qualitative similar protein pattern but also quantitative variations with different intensity bands depending on the extraction method used. The selected extraction method added with sonication proved to work extremely well and efficiently and was able to obtain a high-resolution 2-D electrophoresis (2-DE) map. Prominent spots were successfully identified by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry and corresponded to 76 different proteins involved in the main metabolic pathways. The approach allowed to achieve a protein profiling specific for O. oeni from Aglianico wine with numerous characterized protein products corresponding to many different O. oeni genes and associated with main cellular pathways. Further investigations of the 2-DE protein expression profile will provide useful and interesting information on the molecular mechanisms at the protein level responsible for growth and survival of O. oeni in wine.

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Authors and Affiliations

  1. Dipartimento di Scienze, Università degli Studi della Basilicata, Viale dell’Ateneo Lucano 10, 85100, Potenza, Italy

    Caterina Cafaro, Maria Grazia Bonomo, Rocco Rossano, Marilena Larocca & Giovanni Salzano

  2. Doctoral School of “BioEcoSystems and BioTechnology”, Università degli Studi della Basilicata, Viale dell’Ateneo Lucano 10, 85100, Potenza, Italy

    Caterina Cafaro

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  1. Caterina Cafaro
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  2. Maria Grazia Bonomo
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  3. Rocco Rossano
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  4. Marilena Larocca
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  5. Giovanni Salzano
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Correspondence to Maria Grazia Bonomo.

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Cafaro, C., Bonomo, M.G., Rossano, R. et al. Efficient recovery of whole cell proteins in Oenococcus oeni—a comparison of different extraction protocols for high-throughput malolactic starter applications. Folia Microbiol 59, 399–408 (2014). https://doi.org/10.1007/s12223-014-0312-8

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  • DOI: https://doi.org/10.1007/s12223-014-0312-8

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