Optimization of R-(+)-alpha-terpineol production by the biotransformation of R-(+)-limonene

J Ind Microbiol Biotechnol. 2008 Sep;35(9):1061-70. doi: 10.1007/s10295-008-0383-0. Epub 2008 Jun 17.


R-(+)-limonene is an abundant and non-expensive by-product of the citrus industry and is, therefore, a suitable starting material for the production of natural flavor and fragrance compounds. The biotransformation of R-(+)-limonene to R-(+)-alpha-terpineol by Fusarium oxysporum 152b has already been reported, although the influence of the main process parameters on the production has not yet been evaluated. In this paper, a Plackett-Burman screening design was used to define the effects of the medium composition (glucose, peptone, yeast extract, malt extract and pH), the presence of a co-substrate (biosurfactant), the cultivation conditions (temperature, agitation), the substrate concentration and the inoculum/culture medium ratio on the absolute amount of R-(+)-alpha-terpineol resulting from this biotransformation. The process conditions were further optimized applying response surface methodology (RSM). The volatiles were extracted using a SPME device and were subsequently quantified by GC-FID and identified by GC-MS. The best results were obtained using 0.5% (v/m) R-(+)-limonene in pure distilled water as the culture medium with an inoculum/culture medium ratio of 0.25 (m/m) and 72 h cultivation at 26 degrees C/240 rpm. Under these conditions the concentration of R-(+)-alpha-terpineol in the culture medium reached 2.4 g L(-1), a production almost six times greater than in earlier trials. The presence of a biosurfactant (0-500 mg L(-1)) did not significantly increase the yield.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Biotransformation
  • Culture Media
  • Cyclohexane Monoterpenes
  • Cyclohexenes / chemical synthesis*
  • Cyclohexenes / metabolism*
  • Food Technology
  • Fusarium / metabolism*
  • Least-Squares Analysis
  • Limonene
  • Monoterpenes / chemical synthesis*
  • Temperature
  • Terpenes / metabolism*


  • Culture Media
  • Cyclohexane Monoterpenes
  • Cyclohexenes
  • Monoterpenes
  • Terpenes
  • alpha-terpineol
  • Limonene