Purification of micrococcal nuclease for use in ribosomal profiling of high-salinity extremophiles.

ABSTRACT: Nucleases, that is, enzymes that catalyze the hydrolysis of phosphodiester bonds in nucleic acids, are essential tools in molecular biology and biotechnology. Staphylococcus aureus nuclease is particularly interesting due to its thermostability and Ca²⁺ dependence, making it the prime choice for applications where nuclease modulation is critical, such as ribosome profiling in bacteria and halophilic archaea. The latter poses a technical and economical challenge: high salt reaction conditions are essential for maintaining ribosome integrity but negatively impact the micrococcal nuclease (MNase) activity, necessitating using large amounts of nuclease to achieve efficient cleavage. Here, we set out to generate an optimized production protocol for two forms of MNase-fully processed

SUBMITTER: Gregorova P 

PROVIDER: S-EPMC11719836 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature