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Optimization of Expression Conditions Enhances Production of Sepiapterin, a Precursor for Tetrahydrobiopterin Biosynthesis, in Recombinant Escherichia coli
Optimization of Expression Conditions Enhances Production of Sepiapterin, a Precursor for Tetrahydrobiopterin Biosynthesis, in Recombinant Escherichia coli
Journal of Microbiology and Biotechnology. 2015. Oct, 25(10): 1709-1713
Copyright © 2015, The Korean Society For Microbiology And Biotechnology
  • Received : June 15, 2015
  • Accepted : July 18, 2015
  • Published : October 28, 2015
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About the Authors
Eun-Hee Park
Department of Food Science and Biotechnology, Kangwon National University, Chuncheon 200-701, Republic of Korea
Won-Heong Lee
Department of Bioenergy Science and Technology, Chonnam National University, Gwangju 500-757, Republic of Korea
Mi-Hee Jang
Department of Food Science and Biotechnology, Kangwon National University, Chuncheon 200-701, Republic of Korea
Myoung-Dong Kim
Department of Food Science and Biotechnology, Kangwon National University, Chuncheon 200-701, Republic of Korea
mdkim@kangwon.ac.kr

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 T 7 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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Acknowledgements
This study was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2009-0066557). M.D. Kim was supported by “Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ009477),” Rural Development Administration, Republic of Korea.
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