Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Optimization of Expression Conditions Enhances Production of Sepiapterin, a Precursor for Tetrahydrobiopterin Biosynthesis, in Recombinant Escherichia coli

  • Park, Eun-Hee (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Lee, Won-Heong (Department of Bioenergy Science and Technology, Chonnam National University) ;
  • Jang, Mi-Hee (Department of Food Science and Biotechnology, Kangwon National University) ;
  • Kim, Myoung-Dong (Department of Food Science and Biotechnology, Kangwon National University)
  • Received : 2015.06.15
  • Accepted : 2015.07.18
  • Published : 2015.10.28
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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℃ 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℃ and 1 mM IPTG).

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