Expression of a Recombinant Cholesterol Esterase from Mustela putorius furo in Pichia pastoris and Its Applicability for γ-Oryzanol Hydrolysis

J Agric Food Chem. 2024 Oct 2;72(39):21702-21710. doi: 10.1021/acs.jafc.4c05238. Epub 2024 Sep 19.

Abstract

Ferulic acid (FA) exhibits antioxidant and anti-inflammatory properties, making it valuable for numerous industrial applications. Traditionally, FA is produced by the alkaline hydrolysis of γ-oryzanol, which is typically associated with wastewater generation. Recently, an increasing demand of natural FA necessitates its green production via enzymatic hydrolysis of γ-oryzanol, a mixture comprising triterpene alcohol ferulates and phytosteryl ferulates. Thus far, γ-oryzanol can be hydrolyzed by only four commercial cholesterol esterases with low yields. Herein, we report a recombinant cholesterol esterase from Mustela putorius furo (MPFCE) for the enzymatic hydrolysis of γ-oryzanol. The enzyme yielded 25.5% FA, which is the highest reported through enzymatic means thus far. The hydrolysis profile revealed that the enhanced yield primarily resulted from the near-complete hydrolysis of phytosteryl ferulates, together with slight hydrolysis of triterpene alcohol ferulates. MPFCE serves as a potential candidate for the enzymatic production of FA through targeted hydrolysis of γ-oryzanol.

Keywords: biocatalysis; enzymatic hydrolysis; ferulic acid; steryl ferulate; γ-oryzanol.

MeSH terms

  • Coumaric Acids / chemistry
  • Coumaric Acids / metabolism
  • Gene Expression
  • Hydrolysis
  • Phenylpropionates* / chemistry
  • Phenylpropionates* / metabolism
  • Plant Proteins / chemistry
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Saccharomycetales / enzymology
  • Saccharomycetales / genetics
  • Saccharomycetales / metabolism
  • Sterol Esterase* / chemistry
  • Sterol Esterase* / genetics
  • Sterol Esterase* / metabolism

Substances

  • Phenylpropionates
  • gamma-oryzanol
  • Sterol Esterase
  • Recombinant Proteins
  • Coumaric Acids
  • ferulic acid
  • Plant Proteins

Supplementary concepts

  • Komagataella pastoris