Optimization of thermophilic trans-isoprenyl diphosphate synthase expression in Escherichia coli by response surface methodology

Biotechnol Appl Biochem. 2017 Jan;64(1):70-78. doi: 10.1002/bab.1459. Epub 2016 Mar 14.


We optimized the heterologous expression of trans-isoprenyl diphosphate synthase (IDS), the key enzyme involved in the biosynthesis of trans-polyisoprene. trans-Polyisoprene is a particularly valuable compound due to its superior stiffness, excellent insulation, and low thermal expansion coefficient. Currently, trans-polyisoprene is mainly produced through chemical synthesis and no biotechnological processes have been established so far for its large-scale production. In this work, we employed D-optimal design and response surface methodology to optimize the expression of thermophilic enzymes IDS from Thermococcus kodakaraensis. The design of experiment took into account of six factors (preinduction cell density, inducer concentration, postinduction temperature, salt concentration, alternative carbon source, and protein inhibitor) and seven culture media (LB, NZCYM, TB, M9, Ec, Ac, and EDAVIS) at five different pH points. By screening only 109 experimental points, we were able to improve IDS production by 48% in close-batch fermentation.

Keywords: design of experiments; protein expression optimization; response surface methodology; thermophilic enzymes; trans-isoprenyl diphosphate synthase.

MeSH terms

  • Alkyl and Aryl Transferases* / biosynthesis
  • Alkyl and Aryl Transferases* / chemistry
  • Alkyl and Aryl Transferases* / genetics
  • Archaeal Proteins* / biosynthesis
  • Archaeal Proteins* / chemistry
  • Archaeal Proteins* / genetics
  • Escherichia coli* / chemistry
  • Escherichia coli* / genetics
  • Escherichia coli* / metabolism
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Thermococcus / enzymology
  • Thermococcus / genetics*


  • Archaeal Proteins
  • Recombinant Proteins
  • Alkyl and Aryl Transferases