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Firmware update download options for Intenso TAB 814.Investigation of lysates prepared from Pichia pastoris by HPLC-DAD-MSn and HPLC-DAD-FLD: metabolomics approach to identify a P. pastoris mutant strain suitable for the secretion of green fluorescent protein.
In this paper, HPLC-DAD-MSn and HPLC-DAD-FLD were used to analyze lysates prepared from Pichia pastoris (strain GS115-GCGGCTA) cells grown in basic medium, containing wild-type (WT) and mutant (mut) GFP proteins, to identify a P. pastoris strain suitable for the expression and secretion of a heterologous protein. The data confirmed the unexpected cell-dependent differences in the P. pastoris protein expression profiles of lysates prepared from cells grown on methanol as sole carbon source. Lysates derived from mut cells grown on basic medium had higher levels of a few proteins that are normally not present in P. pastoris cells grown on methanol. The use of HPLC-DAD-MSn and HPLC-DAD-FLD allowed the identification of proteins that contain and post-translationally modify fluorescent tags, showing high specificity for the identification of the various mutants. The results confirmed that lysates prepared from P. pastoris cells grown on basic medium are suitable for detailed proteomic analysis of secreted proteins, providing not only the information on the presence of secreted proteins but also that of proteins that modified the tag, in this case the fluorescent tag.Q:
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