| Abstract [eng] |
β-carbonic anhydrase (β-CA) is an enzyme, which has a unique feature - is able to catalyze reversible CO2 hydration reaction. The fixed atmospheric CO2 can be converted into high-value products: acrylates, methane, polyurethane, etc. This study was focused on examination of various yeast genus for efficient heterologous expression and secretion of β-CA from bacteria Bacillus mojavensis. For the first time, the yeasts were shown to be a promising host for efficient β-CA protein production. The investigation was carried out with Saccharomyces cerevisiae and Pichia pastoris yeast strains. Vector pFX7αF-β-cab was used in Saccharomyces cerevisiae expression system. The targeted protein was produced efficiently, albeit majority of the recombinant enzyme was in the insoluble form. The two shuttle vectors pPIC9K-β-cab and pPIC3.5K-β-cab, which were constructed by principle of homologous recombination in bacteria E.coli, were applied in Pichia pastoris expression system. In P. pastoris expression system (pPIC3.5K), β-CA was efficiently synthesised in the soluble form. The heterologous protein had esterase and hydratase activities. In the second P. pastoris expression system (pPIC9K), the majority of β-CA was produced in the insoluble form; nevertheless, the part of synthesised targeted protein was detected in extracellular matrix by the protonogram method. |
