title: Protein aggregation and membrane lipid modifications under lactic acid stress in wild type and OPI1 deleted Saccharomyces cerevisiae strains.
abstract: BACKGROUND
  Lactic acid is a versatile chemical platform with many different industrial applications. Yeasts have been demonstrated as attractive alternative to natural lactic acid producers since they can grow at low pH, allowing the direct purification of the product in the desired acidic form. However, when very high concentrations of organic acids are reached, the major limitation for a viable production is the toxic effect of the product. The accumulation in the cytosol of H(+) and of the weak organic counter-anions triggers a cellular reprogramming. Here, the effects of lactic acid exposure on Saccharomyces cerevisiae have been evaluated by Fourier transform infrared (FTIR) microspectroscopy. In addition to -omic techniques, describing these responses in terms of systems and networks, FTIR microspectroscopy allows a rapid acquisition of the cellular biochemical fingerprint, providing information on the major classes of macromolecules.
  
  
  RESULTS
  FTIR analyses on Saccharomyces cerevisiae cells under lactic acid stress at low pH revealed some still uncharacterized traits: (1) a direct correlation between lactic acid exposure and a rearrangement in lipid hydrocarbon tails, together with a decrease in the signals of phosphatidylcholine (PC), one of the main components of cell membrane; (2) a rearrangement in the cell wall carbohydrates, including glucans and mannans (3) a significant yet transient protein aggregation, possibly responsible for the observed transient decrease of the growth rate. When repeated on the isogenic strain deleted in OPI1, encoding for a transcriptional repressor of genes involved in PC biosynthesis, FTIR analysis revealed that not only the PC levels were affected but also the cell membrane/wall composition and the accumulation of protein aggregates, resulting in higher growth rate in the presence of the stressing agent.
  
  
  CONCLUSIONS
  This work revealed novel effects evoked by lactic acid on cell membrane/wall composition and protein aggregation in S. cerevisiae cells. We consequently demonstrated that the targeted deletion of OPI1 resulted in improved lactic acid tolerance. Considering that stress response involves many and different cellular networks and regulations, most of which are still not implemented in modelling, these findings constitute valuable issues for interpreting cellular rewiring and for tailoring ameliorated cell factories for lactic acid production.
authors.affiliation: Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, Milan, 20126, Italy. n.berterame1@campus.unimib.it.
authors.forename: Nadia Maria
authors.initials: NM
authors.lastname: Berterame
authors.affiliation: Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, Milan, 20126, Italy. danilo.porro@unimib.it.|SYSBIO - Centre of Systems Biology, Milano and Roma, Italy. danilo.porro@unimib.it.
authors.forename: Danilo
authors.initials: D
authors.lastname: Porro
authors.affiliation: Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, Milan, 20126, Italy. diletta.ami@unimib.it.|Department of Physics, University of Milano-Bicocca, Piazza della Scienza 3, Milan, 20126, Italy. diletta.ami@unimib.it.|Consorzio Nazionale Interuniversitario per le Scienze fisiche della Materia (CNISM) UdR Milano-Bicocca, Milan, 20126, Italy. diletta.ami@unimib.it.
authors.forename: Diletta
authors.initials: D
authors.lastname: Ami
authors.affiliation: Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, Milan, 20126, Italy. paola.branduardi@unimib.it.
authors.forename: Paola
authors.initials: P
authors.lastname: Branduardi
mesh_terms: D000222:Adaptation, Physiological / Q000187:drug effects; D017353:Gene Deletion; D019344:Lactic Acid / Q000633:toxicity*; D015227:Lipid Peroxidation / Q000187:drug effects; D008563:Membrane Lipids / Q000378:metabolism*; D066329:Protein Aggregates / Q000187:drug effects*; D012097:Repressor Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared; D013312:Stress, Physiological / Q000187:drug effects*; D056811:Unfolded Protein Response / Q000187:drug effects
chemical_list: D008563:Membrane Lipids; C096105:OPI1 protein, S cerevisiae; D066329:Protein Aggregates; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D019344:Lactic Acid