Objective
The present project aims to disclose the previously unknown structural bases of proteins adapted to alkaline and acid environments. This will be accomplished by a coordinated investigation of a number of key parameters of model proteins from pH-extremophilic microbes with a representative selection of experimental and theoretical methods governed by three expert teams from NIS and two from INTAS countries.
The research will focus on transaminases from Bacillus species having high and low pH-optima of growth.
The enzymes isolated from Bacillus and recombinant E. coli cells will be used for extensive enzymological and physico-chemical characterisation. To analyse folding and stability, transaminases and their mutants will be investigated under increasingly denaturing solvent conditions (variable pH, temperature and denaturant concentration) by a combination of spectroscopic and thermodynamic approaches. An extensive research on kinetics will be performed to characterise the stability of tertiary and quaternary structure of transaminases through kinetic parameters. Spatial structures of the proteins will be elucidated by crystallographic studies. The experimental results will be supplemented with computer model building to illustrate the differences between alkaliphilic, neutrophilic and acidophilic enzymes. The secondary, tertiary, and quaternary interactions that contribute to folding and stability of unique spatial structure of the transaminases from pH-extremophiles will be compared with stabilisation factors revealed for other extremophiles.
The results will expand the knowledge of the very bases of protein chemistry, and may be exploited in constructing of enzymes stable and active at high or low pH for the needs of medicine, industry, agriculture and other areas
Topic(s)
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20520 Turku
Finland