Expression of a\nRecombinant Cholesterol Esterase from Mustela putorius\nfuro in Pichia pastoris and Its Applicability for γ‑Oryzanol Hydrolysis

Ferulic acid (FA) exhibits antioxidant and anti-inflammatory\nproperties,\nmaking it valuable for numerous industrial applications. Traditionally,\nFA is produced by the alkaline hydrolysis of <i>γ</i>-oryzanol, which is typically associated with wastewater generation.\nRecently, an increasing demand of natural FA necessitates its green\nproduction via enzymatic hydrolysis of <i>γ</i>-oryzanol,\na mixture comprising triterpene alcohol ferulates and phytosteryl\nferulates. Thus far, <i>γ</i>-oryzanol can be hydrolyzed\nby only four commercial cholesterol esterases with low yields. Herein,\nwe report a recombinant cholesterol esterase from Mustela\nputorius furo (MPFCE) for the enzymatic hydrolysis\nof <i>γ</i>-oryzanol. The enzyme yielded 25.5% FA,\nwhich is the highest reported through enzymatic means thus far. The\nhydrolysis profile revealed that the enhanced yield primarily resulted\nfrom the near-complete hydrolysis of phytosteryl ferulates, together\nwith slight hydrolysis of triterpene alcohol ferulates. MPFCE serves\nas a potential candidate for the enzymatic production of FA through\ntargeted hydrolysis of <i>γ</i>-oryzanol.