Registration Number
0223U005353,
0123U103550
, R & D reports
Title
Mechanisms of specific autophagic degradation of organelles and cytosolic enzymes in yeast as model system of human diseases
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Head
Sybirnyi Andrii A., Доктор біологічних наук
Registration Date
20-12-2023
Organization
Institute of Cell Biology of the National Academy of Sciences of Ukraine
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The development of new approaches to the study of the mechanisms of degradation of peroxisomes and cytosolic proteins in the methylotrophic yeast K. phaffii, as well as the study of the mechanisms of degradation of both homo- and heterologous proteins creates prerequisites for the construction of effective yeast-based producers of target proteins.
The selection system using the β-galactosidase-GFP chimeric protein allows to study the processes of degradation of cytosolic proteins by a wider range of methods: biochemical determination of the activity of the β-galactosidase enzyme and visual determination due to the fluorescent tag GFP allow to find out in which cell compartments the processes of splitting this protein take place.
For the development of protein producers of industrial interest, minimizing the level of target protein degradation in the cytosol is critical. Fld1 and Fdh1 serve as convenient model proteins for studying cytosolic protein degradation in methylotrophic yeast. This is explained by their participation in the utilization of methanol. Methanol is oxidized to formaldehyde by AOX, which is then converted to formate by Fld1, and finally, Fdh1 catalyzes the oxidation of formate to carbon dioxide. The specific activity of Fld1 and Fdh1 rapidly decreases when methylotrophically grown cells are transferred to a medium with glucose. As a result of the conducted experiments, it was shown that the degradation of Fld1 and Fdh1 takes place by the vacuolar pathway, and a platform was created for the construction of efficient producers of heterologous proteins, which are of great biotechnological importance.
Product Description
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Dmytruk Kostyantyn V.
Dmytruk Olena V.
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2023-12-20
