Validation and stabilization of a prophage lysin of Clostridium perfringens by yeast surface display and co-evolutionary models.

Validation and stabilization of a prophage lysin of Clostridium perfringens by yeast surface display and co-evolutionary models. Appl Environ Microbiol. 2019 Mar 08;: Authors: Ritter SC, Hackel BJ Abstract Bacteriophage lysins are compelling antimicrobial proteins whose biotechnological utility and evolvability would be aided by elevated stability. Lysin catalytic domains, which evolved as modular entities distinct from cell wall binding domains, can be classified into one of several families with highly conserved structure and function, many of which contain thousands of annotated homologous sequences. Motivated by the quality of this evolutionary data, the performance of generative protein models incorporating co-evolutionary information was analyzed to predict the stability of variants in a collection of 9,749 multi-mutants across 10 libraries diversified at different regions of a putative lysin from a prophage region of a Clostridium perfringens genome. Protein stability was assessed via a yeast surface display assay with accompanying high-throughput sequencing. Statistical fitness of mutant sequences, derived from second-order Potts models inferred with different levels of sequence homolog information, was predictive of experimental stability with AUCs ranging from 0.78 to 0.85. To extract an experimentally derived model of stability, a logistic model with site-wise score contributions was regressed on the collection of multi-mu...
Source: Applied and Environmental Microbiology - Category: Microbiology Authors: Tags: Appl Environ Microbiol Source Type: research