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KMID : 0545120150250101709
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Journal of Microbiology and Biotechnology
2015 Volume.25 No. 10 p.1709 ~ p.1713
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Optimization of Expression Conditions Enhances Production of Sepiapterin, a Precursor for Tetrahydrobiopterin Biosynthesis, in Recombinant Escherichia coli
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Park Eun-Hee
Lee Won-Heong
Jang Mi-Hee
Kim Myoung-Dong
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Abstract
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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).
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KEYWORD
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Sepiapterin, GTP cyclohydrolase 1 (GCH1), 6-pyruvoyltetrahydropterin synthase (PTPS), Escherichia coli
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