Optimizated Prolyl‐4‐Hydroxylase Expression in Komagataella phaffii Enables Efficient Triple‐Helix Formation of Recombinant Human Collagen

U/L. Additional improvements via co-expression of molecular chaperones and fusion with solubility-enhancing tags elevated activity to 1.35 × 10⁴ U/L. Finally, co-expression of the BaP4H with rhCol enabled the successful formation of the triple-helical structure, which was not observed with rhCol expressed alone. This study establishes an effective strategy for optimizing P4H expression and enables the functional biosynthesis of recombinant collagen, with promising potential for biomedical and industrial applications.