Studies on the Structure and Properties of Membrane Phospholipase A₁ Inclusion Bodies Formed at Low Growth Temperatures Using GFP Fusion Strategy.

ABSTRACT: The effect of cultivation temperatures (37, 26, and 18 °C) on the conformational quality of Yersinia pseudotuberculosis phospholipase A₁ (PldA) in inclusion bodies (IBs) was studied using green fluorescent protein (GFP) as a folding reporter. GFP was fused to the C-terminus of PldA to form the PldA-GFP chimeric protein. It was found that the maximum level of fluorescence and expression of the chimeric protein is observed in cells grown at 18 °C, while at 37 °C no formation of fluorescently active forms of PldA-GFP occurs. The size, stability in denaturant solutions, and enzymatic and biological activity of PldA-GFP IBs expressed at 18 °C, as well as the secondary structure and arrangement of protein molecules inside the IBs, were studied. Solubilization of the chimeric pr

SUBMITTER: Bakholdina SI 

PROVIDER: S-EPMC8271855 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature