Cells of Candida utilis for in vitro (R)-phenylacetylcarbinol production in an aqueous/octanol two-phase reactor

Biotechnol Lett. 2005 Apr;27(8):575-81. doi: 10.1007/s10529-005-3252-1.

Abstract

(R)-Phenylacetylcarbinol (PAC), a pharmaceutical precursor, was produced from benzaldehyde and pyruvate by pyruvate decarboxylase (PDC) of Candida utilis in an aqueous/organic two-phase emulsion reactor. When the partially purified enzyme in this previously established in vitro process was replaced with C. utilis cells and the temperature was increased from 4 to 21 degrees C, a screen of several 1-alcohols (C4-C9) confirmed the suitability of 1-octanol as the organic phase. Benzyl alcohol, the major by-product in the commercial in vivo conversion of benzaldehyde and sugar to PAC by Saccharomyces cerevisiae, was not formed. With a phase volume ratio of 1:1 and 5.6 g C. utilis l-1 (PDC activity 2.5 U ml-1), PAC levels of 103 g l-1 in the octanol phase and 12.8 g l-1 in the aqueous phase were produced in 15 h at 21 degrees C. In comparison to our previously published process with partially purified PDC in an aqueous/octanol emulsion at 4 degrees C, PAC was produced at a 4-times increased specific rate (1.54 versus 0.39 mg U-1 h-1) with simplified catalyst production and reduced cooling cost. Compared to traditional in vivo whole cell PAC production, the yield on benzaldehyde was 26% higher, the product concentration increased 3.9-fold (or 6.9-fold based on the organic phase), the productivity improved 3.1-fold (3.9 g l-1 h-1) and the catalyst was 6.9-fold more efficient (PAC/dry cell mass 10.3 g g-1).

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetone / analogs & derivatives*
  • Acetone / analysis
  • Acetone / metabolism
  • Benzaldehydes / metabolism
  • Bioreactors / microbiology*
  • Biotransformation
  • Candida / cytology
  • Candida / drug effects
  • Candida / metabolism*
  • Cell Division / drug effects
  • Chromatography, High Pressure Liquid
  • Glucose / pharmacokinetics
  • Heptanol / chemistry
  • Heptanol / pharmacology
  • Octanols / chemistry
  • Octanols / pharmacology
  • Pyruvic Acid / metabolism

Substances

  • Benzaldehydes
  • Octanols
  • Acetone
  • Pyruvic Acid
  • Heptanol
  • 1-hydroxy-1-phenyl-2-propanone
  • Glucose
  • benzaldehyde