A novel structurally characterized haloacid dehalogenase superfamily phosphatase from Thermococcus thioreducens with diverse substrate specificity

Mgr. Petra HAVLÍČKOVÁ

Advanced ('rigorózní') thesis

Abstract:

This RNDr. thesis is focused on complex studies of a haloacid dehalogenase superfamily phosphatase member with an unknown substrate specificity, isolated from a hyperthermofilic archeon Thermococcus thioreducens. The HAD phosphatase, internally named as Tt82, was isolated, expressed and purified using standard techniques and the crystal structure was solved by X-ray crystallography method. The refined crystal structure displayed characteristic HAD superfamily elements, such as the HAD canonical fold with the HAD signature motifs in the active site. Further, possible substrates for the enzyme were proposed and examined by computational docking in the active site of the HAD phosphatase. The results of the docking studies served as a basis for an experimental measurement of Tt82 enzymatic activity against selected substrates at two different temperatures. HAD phosphatase Tt82 is a promising target for the biotechnological field, mainly for its wide substrate specificity and higher temperature optima.

Language used: English

Date on which the thesis was submitted / produced: 31. 1. 2024

Thesis defence

UNIVERSITY OF SOUTH BOHEMIA IN ČESKÉ BUDĚJOVICE

Faculty of Science

Advanced ('rigorózní řízení') programme / field:
Biochemie / Biochemie