Abstract The aim of this study was to demonstrate the potential of PEF for inducing autolysis of R. glutinis, with the purpose of designing a more efficient and ecofriendly carotenoid… Click to show full abstract
Abstract The aim of this study was to demonstrate the potential of PEF for inducing autolysis of R. glutinis, with the purpose of designing a more efficient and ecofriendly carotenoid extraction process: an extraction from fresh biomass, using cheaper, non-toxic, environmental-friendly solvents. Propidium iodide uptake and release of intracellular components revealed the irreversible electroporation of R. glutinis by PEF. Flow cytometry measurements detected morphological changes in PEF-treated R. glutinis cells during incubation caused by the autolysis triggering effect of electroporation. After submitting the fresh biomass to a PEF treatment (15 kV/cm, 150 μs) that irreversibly electroporated more than the 90% of the cells, ethanol proved ineffective for extracting carotenoids from fresh biomass of R. glutinis. However, after incubating the PEF-treated fresh biomass for 24 h at 20 °C in a pH 7 buffer, ca. 240 μg/g d.w. of carotenoids were recovered after 1 h of extraction in ethanol. The highest amount of carotenoids extracted (375 μg/g d.w.) from the PEF-treated cells of R. glutinis was obtained after having incubated them at 25 °C for 24 h in a medium of pH 8.0. The improvement in carotenoid extraction by incubating the R. glutinis cells after PEF treatment seems to be caused by PEF-triggered autolysis, which tends to disrupt the association of carotenoids with other molecules present in the cytoplasm, and causes a degradation of the cell wall.
               
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