ABSTRACT
Halophilic aminopeptidases with broad substrate specificity represent valuable biocatalysts for promoting protein hydrolysis in high-salt fermented foods. In this study, an M42 aminopeptidase from the halophilic archaeon, Haladaptatus litoreus, was identified and designated as APHap. The optimal reaction conditions for APHap were 2–2.5 M NaCl, a temperature of 60°C, and a pH of 7.5. It displayed robust activity and stability across a wide range of salinity, temperature, and pH conditions. APHap demonstrated exceptional tolerance to both organic solvents and surfactants. In contrast to most characterized M42 aminopeptidases, APHap exhibited a broad substrate spectrum, with the highest activity observed when using Leu-p-nitroaniline as the substrate. The Vmax and Km for APHap were 5.28 µmol/min/mg and 0.59 mM, respectively. When applied for fish protein hydrolysis in hypersaline conditions, APHap significantly increased the total free amino acid content, particularly enhancing the proportion of sweet and umami amino acids. To our knowledge, APHap is the first halophilic and mesophilic M42 aminopeptidase characterized from the genus Haladaptus. These desirable properties indicated that APHap has great potential for enhancing protein hydrolysis during the processing of high-salt fermented foods.
Practical Application
The characteristics of APHap conform with the demands of high-salt fermented food production, highlighting its potential as a biocatalyst for improving both process efficiency and product quality.