Biotransformation of limonene by an endophytic fungus using synthetic and orange residue-based media

Fungal Biol. 2017 Feb;121(2):137-144. doi: 10.1016/j.funbio.2016.11.003. Epub 2016 Nov 30.


Aroma and fragrances have high commercial value for use in food, cosmetics and perfumes. The biotransformation of terpenes by microorganisms represents an attractive alternative method for production of flavourings. Endophytic fungi offer a great potential for the production of several groups of compounds; however, few studies have evaluated the biotransformation of limonene. Following preliminary studies on the biotransformation of limonene, submerged fermentation was carried out using an endophytic fungus isolated from Pinus taeda and identified as Phomopsis sp. The presence of several biotransformation products was detected and identified by mass spectrometry (GC-MS). The studied strain showed a divergent metabolic behaviour, as compounds of interest such as α-terpineol, carvone, and limoneno-1,2-diol were produced under different conditions. In addition to the minor metabolites terpinen-4-ol, menthol and carveol, this strain also produced major metabolites, including 0.536 g L-1 carvone and 2.08 g L-1 limonene-1,2-diol in synthetic medium and 2.10 g L-1 limonene-1,2-diol in a natural orange extract medium with single fed-batch, while the cyclic fed-batch resulted in concentrations less than 1 g L-1. Therefore, our study produced a wide variety of limonene derivatives at a high concentration using a natural medium and a newly isolated endophytic fungal strain.

Keywords: Aroma; Endophyte; Limonene-1,2-diol; Submerged fermentation; Terpenoids.

MeSH terms

  • Ascomycota / classification
  • Ascomycota / isolation & purification
  • Ascomycota / metabolism*
  • Biotransformation
  • Culture Media / chemistry
  • Cyclohexenes / metabolism*
  • Endophytes / classification
  • Endophytes / isolation & purification
  • Endophytes / metabolism*
  • Flavoring Agents / metabolism
  • Gas Chromatography-Mass Spectrometry
  • Limonene
  • Pinus taeda / microbiology
  • Sequence Analysis, DNA
  • Terpenes / metabolism*


  • Culture Media
  • Cyclohexenes
  • Flavoring Agents
  • Terpenes
  • Limonene