Phosphatases of the haloacid dehalogenase superfamily are one of the cell's tools for dephosphorylation of many diverse endogenous and exogenous compounds. This work is aimed at enzymes Tt82 and cytosolic purine 5'-nucleotidase II (cN-II), two members of this large enzyme superfamily. The Tt82 originates in the hyperthermophilic archaeon Thermococcus thioreducens. Up to date, there is only a small amount of knowledge about properties and biological function of this enzyme. Based on its sequence and structure, it was predicted that the Tt82 should possess a phosphatase catalytic activity. Consequently, potential substrates of the Tt82 were proposed by the molecular docking. In this work, the phosphatase activity of the Tt82 was confirmed together with several of its substrates: AMP, D-glucose 1-phosphate, D-glucose 6-phosphate and p-nitrophenyl phosphate (pNPP). Activity towards AMP and pNPP was then characterized by steady-state kinetics at 37 řC and 60 řC. In consistence with its thermophilic origin, the Tt82 showed markedly higher activity towards both substrates at 60 řC. Nonetheless, the effectivity of the Tt82 catalytic activity towards these substrates was actually very low. This leads to assumption, that the identified substrates are probably not biologically relevant. On the other hand, it is quite...
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:434093 |
| Date | January 2020 |
| Creators | Brinsa, Vítězslav |
| Contributors | Maloy Řezáčová, Pavlína, Vaněk, Ondřej |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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