Biosynthesis of Sandalwood Oil: Santalum album CYP76F cytochromes P450 produce santalols and bergamotol

PLoS One. 2013 Sep 18;8(9):e75053. doi: 10.1371/journal.pone.0075053. eCollection 2013.


Sandalwood oil is one of the world's most highly prized essential oils, appearing in many high-end perfumes and fragrances. Extracted from the mature heartwood of several Santalum species, sandalwood oil is comprised mainly of sesquiterpene olefins and alcohols. Four sesquiterpenols, α-, β-, and epi-β-santalol and α-exo-bergamotol, make up approximately 90% of the oil of Santalum album. These compounds are the hydroxylated analogues of α-, β-, and epi-β-santalene and α-exo-bergamotene. By mining a transcriptome database of S. album for candidate cytochrome P450 genes, we cloned and characterized cDNAs encoding a small family of ten cytochrome P450-dependent monooxygenases annotated as SaCYP76F37v1, SaCYP76F37v2, SaCYP76F38v1, SaCYP76F38v2, SaCYP76F39v1, SaCYP76F39v2, SaCYP76F40, SaCYP76F41, SaCYP76F42, and SaCYP76F43. Nine of these genes were functionally characterized using in vitro assays and yeast in vivo assays to encode santalene/bergamotene oxidases and bergamotene oxidases. These results provide a foundation for production of sandalwood oil for the fragrance industry by means of metabolic engineering, as demonstrated with proof-of-concept formation of santalols and bergamotol in engineered yeast cells, simultaneously addressing conservation challenges by reducing pressure on supply of sandalwood from native forests.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • Cytochrome P-450 Enzyme System / chemistry
  • Cytochrome P-450 Enzyme System / genetics
  • Cytochrome P-450 Enzyme System / metabolism*
  • DNA, Complementary / genetics
  • Gas Chromatography-Mass Spectrometry
  • Gene Expression
  • Isoenzymes
  • Kinetics
  • Phylogeny
  • Plant Oils / chemistry
  • Plant Oils / metabolism*
  • Plant Proteins / chemistry
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Polycyclic Sesquiterpenes
  • Santalum / classification
  • Santalum / genetics
  • Santalum / metabolism*
  • Sesquiterpenes / chemistry
  • Sesquiterpenes / metabolism*
  • Substrate Specificity
  • Yeasts / genetics
  • Yeasts / metabolism


  • DNA, Complementary
  • Isoenzymes
  • Plant Oils
  • Plant Proteins
  • Polycyclic Sesquiterpenes
  • Sesquiterpenes
  • santalol
  • Cytochrome P-450 Enzyme System
  • sandalwood oil

Grant support

Funding for this work was provided by Allyix Inc. (to JB), the Natural Sciences and Engineering Research Council of Canada (to JB), and the Australian Research Council and the Forest Products Commission of Western Australia through Linkage Project LP0882690 (to JAP, ELG, ELB, JB). JB was supported in part by the UBC Distinguished University Scholar program. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.