Enzyme stabilisation due to incorporation of a fluorinated non-natural amino acid at the protein surface.

ABSTRACT: We have previously engineered E. coli transketolase (TK) enzyme variants that accept new substrates such as aliphatic or aromatic aldehydes, and also with improved thermal stability. Irreversible aggregation is the primary mechanism of deactivation for TK in the buffers used for biocatalysis, and so we were interested in determining the extent to which this remains true in more complex media, crude cell lysates or even in vivo. Such understanding would better guide future protein engineering efforts. NMR offers a potential approach to probe protein structure changes, aggregation, and diffusion, and¹⁹F-labelled amino acids are a useful NMR probe for complex systems with little or no background signal from the rest of the protein or their environment. Here we labelled E. coli TK w

SUBMITTER: Mukhopadhyay A 

PROVIDER: S-EPMC11564776 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature