Superanionic Solvent-Free Liquid Enzymes Exhibit Enhanced Structures and Activities

The surface of a carboxylate-enriched octuple mutant of Bacillus subtilis lipase A (8M) is chemically anionized to produce core (8M)-shell (cationic polymer surfactants) bionanoconjugates in protein liquid form, which are termed anion-type biofluids. The resultant lipase biofluids exhibit a 2.5-fold increase in hydrolytic activity when compared with analogous lipase biofluids based on anionic polymer surfactants. In addition, the applicability of the anion-type biofluid using Myoglobin (Mb) that is well studied in anion-type solvent-free liquid proteins is evaluated. Although anionization resulted in the complete unfolding of Mb, the active alpha-helix level is partially recovered in the anion-type biofluids, and the effect is accentuated in the cation-type Mb biofluids. These highly active anion-type solvent-free liquid enzymes exhibit increased thermal stability and provide a new direction in solvent-free liquid protein research.

Automated summary

This study focuses on the anionization of a carboxylate-enriched mutant of Bacillus subtilis lipase A (8M) to produce biofluids with enhanced hydrolytic activity. The resulting biofluids show a 2.5-fold increase in activity compared to those based on anionic polymer surfactants. Additionally, the use of anion-type biofluids with Myoglobin (Mb) is evaluated, revealing partial recovery of the active alpha-helix level in the biofluids. These highly active solvent-free liquid enzymes exhibit increased thermal stability and open up new possibilities in solvent-free liquid protein research.