1887

Abstract

We characterized the biosynthetic pathway of tetrahydrobiopterin (BH) in the lipid-producing fungus . The BH cofactor is essential for various cell processes, and is probably present in every cell or tissue of higher organisms. Genes encoding two copies of GTP cyclohydrolase I (GTPCH-1 and GTPCH-2) for the conversion of GTP to dihydroneopterin triphosphate (H-NTP), 6-pyruvoyltetrahydropterin synthase (PTPS) for the conversion of H-NTP to 6-pyruvoyltetrahydropterin (PPH), and sepiapterin reductase (SR) for the conversion of PPH to BH, were expressed heterologously in . The recombinant enzymes were produced as His-tagged fusion proteins and were purified to homogeneity to investigate their enzymic activities. Enzyme products were analysed by HPLC and electrospray ionization-MS. Kinetic parameters and other properties of GTPCH, PTPS and SR were investigated. Physiological roles of BH in are discussed, and comparative analyses between GTPCH, PTPS and SR proteins and other homologous proteins were performed. The presence of two functional GTPCH enzymes has, as far as we are aware, not been reported previously, reflecting the unique ability of this fungus to synthesize both BH and folate, using the GTPCH product as a common substrate. To our knowledge, this study is the first to report the comprehensive characterization of a BH biosynthesis pathway in a fungus.

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2011-11-01
2019-10-15
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