Charge Engineering Improves the Performance of Bst DNA Polymerase Fusions.

ABSTRACT: The charge states of proteins can greatly influence their stabilities and interactions with substrates, and the addition of multiple charges (supercharging) has been shown to be a successful approach for engineering protein stability and function. The addition of a fast-folding fusion domain to the Bacillus stearothermophilus DNA polymerase improved its functionality in isothermal amplification assays, and further charge engineering of this domain has increased both protein stability and diagnostics performance. When combined with mutations that stabilize the core of the protein, the charge-engineered fusion domain leads to the ability to carry out loop-mediated isothermal amplification (LAMP) at temperatures up to 74° C or in the presence of high concentrations of urea, with detect

SUBMITTER: Paik I 

PROVIDER: S-EPMC9514385 | biostudies-literature | 2022 Apr

REPOSITORIES: biostudies-literature