Heterologous Production of Cyprosin B in Nicotiana benthamiana: Unveiling the Role of the Plant-Specific Insert Domain in Protein Function and Subcellular Localisation.

ABSTRACT: Plant systems have gained increased attention as an alternative platform for producing heterologous proteins, particularly for industrially relevant proteins. The Cynara cardunculus L. flower extract is traditionally used in cheese production across Mediterranean countries due to its milk-clotting properties. To address the growing demand for plant-based milk-clotting enzymes, we investigated the heterologous production of cyprosin B (CYPB), a key milk-clotting enzyme, in Nicotiana benthamiana. We also examined the role of its plant-specific insert (PSI) domain in enzymatic activity, protein yield and subcellular localisation. Full-length CYPB and a PSI domain-deleted variant (CYPBΔPSI) were transiently expressed in N. benthamiana leaves using agroinfiltration. Proteins were purified 9 day

SUBMITTER: Muthusamy S 

PROVIDER: S-EPMC12854894 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature