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http://dx.doi.org/10.4014/jmb.1506.06034

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)
Publication Information
Journal of Microbiology and Biotechnology / v.25, no.10, 2015 , pp. 1709-1713 More about this Journal
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).
Keywords
Sepiapterin; GTP cyclohydrolase 1 (GCH1); 6-pyruvoyltetrahydropterin synthase (PTPS); Escherichia coli;
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