Optimization of Expression Conditions Enhances Production of Sepiapterin, a Precursor for Tetrahydrobiopterin Biosynthesis, in Recombinant Escherichia coli

J Microbiol Biotechnol. 2015 Oct;25(10):1709-13. doi: 10.4014/jmb.1506.06034.

Abstract

Sepiapterin is a precursor for the synthesis of tetrahydrobiopterin (BH4), which is a wellknown cofactor for aromatic amino acid hydroxylation and nitric oxide synthesis in higher mammals. In this study, a recombinant Escherichia coli BL21(DE3) strain harboring cyanobacterial guanosine 5'-triphosphate cyclohydrolase 1 (GCH1) and human 6- pyruvoyltetrahydropterin synthase (PTPS) genes was constructed to produce sepiapterin. The optimum conditions for T7 promoter-driven expression of GCH1 and PTPS were 30°C and 0.1 mM isopropyl-β-D-thioglucopyranoside (IPTG). The maximum sepiapterin concentration of 88.1 ± 2.4 mg/l was obtained in a batch cultivation of the recombinant E. coli, corresponding to an 18-fold increase in sepiapterin production compared with the control condition (37°C and 1 mM IPTG).

Keywords: 6-pyruvoyltetrahydropterin synthase (PTPS); Escherichia coli; GTP cyclohydrolase 1 (GCH1); Sepiapterin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biopterins / analogs & derivatives*
  • Biopterins / biosynthesis
  • Cyanobacteria / enzymology
  • Cyanobacteria / genetics
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism*
  • Gene Expression
  • Humans
  • Metabolic Engineering / methods*
  • Metabolic Networks and Pathways*
  • Pterins / metabolism*
  • Temperature
  • Transcriptional Activation

Substances

  • Pterins
  • Biopterins
  • sepiapterin
  • sapropterin