Metabolic Engineering of Monoterpene Synthesis in Yeast

Biotechnol Bioeng. 2011 Aug;108(8):1883-92. doi: 10.1002/bit.23129. Epub 2011 Mar 21.

Abstract

Terpenoids are one of the largest and most diverse families of natural compounds. They are heavily used in industry, and the trend is toward engineering modified microorganisms that produce high levels of specific terpenoids. Most studies have focused on creating specific heterologous pathways for sesquiterpenes in Escherichia coli or yeast. We subjected the Saccharomyces cerevisiae ERG20 gene (encoding farnesyl diphosphate synthase) to a set of amino acid mutations in the catalytic site at position K197. Mutated strains have been shown to exhibit various growth rate, sterol amount, and monoterpenol-producing capacities. These results are discussed in the context of the potential use of these mutated strains for heterologous expression of monoterpenoid synthases, which was investigated using Ocimum basilicum geraniol synthase. The results obtained with up to 5 mg/L geraniol suggest a major improvement compared with previous available expression systems like Escherichia coli or yeast strains with an unmodified ERG20 gene that respectively delivered amounts in the 10 and 500 µg/L range or even a previously characterized K197E mutation that delivered amounts in the 1 mg/L range.

MeSH terms

  • Amino Acid Substitution / genetics
  • Gene Expression
  • Genetic Engineering*
  • Geranyltranstransferase / genetics
  • Metabolic Networks and Pathways / genetics*
  • Models, Molecular
  • Monoterpenes / metabolism*
  • Ocimum basilicum / enzymology
  • Ocimum basilicum / genetics
  • Phosphoric Monoester Hydrolases / genetics*
  • Phosphoric Monoester Hydrolases / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics

Substances

  • Monoterpenes
  • Recombinant Proteins
  • Saccharomyces cerevisiae Proteins
  • Geranyltranstransferase
  • Phosphoric Monoester Hydrolases