title: Optimization of Expression Conditions Enhances Production of Sepiapterin, a Precursor for Tetrahydrobiopterin Biosynthesis, in Recombinant Escherichia coli.
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).
authors.affiliation: Department of Food Science and Biotechnology, Kangwon National University, Chuncheon 200-701, Republic of Korea.
authors.forename: Eun-Hee
authors.initials: EH
authors.lastname: Park
authors.affiliation: Department of Bioenergy Science and Technology, Chonnam National University, Gwangju 500-757, Republic of Korea.
authors.forename: Won-Heong
authors.initials: WH
authors.lastname: Lee
authors.affiliation: Department of Food Science and Biotechnology, Kangwon National University, Chuncheon 200-701, Republic of Korea.
authors.forename: Mi-Hee
authors.initials: MH
authors.lastname: Jang
authors.affiliation: Department of Food Science and Biotechnology, Kangwon National University, Chuncheon 200-701, Republic of Korea.
authors.forename: Myoung-Dong
authors.initials: MD
authors.lastname: Kim
mesh_terms: D001708:Biopterins / Q000031:analogs & derivatives* / Q000096:biosynthesis; D000458:Cyanobacteria / Q000201:enzymology / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways; D011622:Pterins / Q000378:metabolism*; D013696:Temperature; D015533:Transcriptional Activation
chemical_list: D011622:Pterins; D001708:Biopterins; C016727:sepiapterin; C003402:sapropterin
keywords: 6-pyruvoyltetrahydropterin synthase (PTPS); Escherichia coli; GTP cyclohydrolase 1 (GCH1); Sepiapterin