Discovery and mechanistic analysis of a novel source protein glutaminase PG5 and its potential application

In this study, we successfully obtained a novel source protein glutaminase PG5 with specific activity of 10.4 U/mg, good tolerance and broad substrate profile through big data retrieval. Structural analysis and site-directed mutagenesis revealed that the catalytic pocket of Mature-PG5 contained a large number of aromatic amino acids and hydrophobic amino acids, and that Ser72 greatly affects the properties of the catalytic pocket and the affinity of PG5 for the substrate. In addition, molecular dynamics analysis revealed that the opening and closing between amino acid residues Gly65 and Thr66 with Cys164 at the catalytic cleft could affect substrate binding and product release. In addition, PG5 effectively improved the solubility of fish myofibrillar proteins under low-salt conditions while enhancing their foaming and emulsification properties. This study offers valuable insights into the catalytic mechanism of PG5, which will contribute to its future directed evolution and application in the food industry. [Display omitted] •A novel protein glutaminase PG5 was obtained through big data mining.•Residue Ser72 in catalytic pocket affect the affinity of PG5 toward substrate.•The opening and closing between Gly65 and Thr66 with Cys164 affect substrate hydrolysis.•PG5 enhanced the solubility and functional properties of fish MPs in low-salt condition.