Direct observation of oxidative stress on the cell wall of Saccharomyces cerevisiae strains with atomic force microscopy.
We imaged pores on the surface of the cell wall of three different industrial strains of Saccharomyces cerevisiae using atomic force microscopy. The pores could be enlarged using 10 mM diamide, an SH residue oxidant that attacks surface proteins. We found that two strains showed signs of oxidative damage via changes in density and diameter of the surface pores. We found that the German strain was resistant to diamide induced oxidative damage, even when the concentration of the oxidant was increased to 50 mM. The normal pore size found on the cell walls of American strains had diameters of about 200 nm. Under conditions of oxidative stress the diameters changed to 400 nm. This method may prove to be a useful rapid screening process (45-60 min) to determine which strains are oxidative resistant, as well as being able to screen for groups of yeast that are sensitive to oxidative stress. This rapid screening tool may have direct applications in molecular biology (transference of the genes to inside of living cells) and biotechnology (biotransformations reactions to produce chiral synthons in organic chemistry.
de Souza Pereira
R
R
Núcleo de Atendimento á Comunidade, Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista (UNESP), Araraquara, SP, Brazil.
Geibel
J
J
D000975:Antioxidants / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002473:Cell Wall / Q000378:metabolism / Q000648:ultrastructure; D003958:Diamide / Q000494:pharmacology; D005656:Fungal Proteins / Q000032:analysis / Q000378:metabolism; D018625:Microscopy, Atomic Force / Q000379:methods*; D008954:Models, Biological; D009249:NADP / Q000378:metabolism; D018384:Oxidative Stress / Q000502:physiology*; D010762:Phosphorylase a / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure*
D000975:Antioxidants; D005656:Fungal Proteins; D017382:Reactive Oxygen Species; D003958:Diamide; D009249:NADP; D010762:Phosphorylase a; D002118:Calcium