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Enzymatic Synthesis of 6R-[U-14C]Tetrahydrobiopterin from [U-14C]GTP

✍ Scribed by K. Hatakeyama; M. Hoshiga; S. Suzuki; H. Kagamiyama

Publisher
Elsevier Science
Year
1993
Tongue
English
Weight
391 KB
Volume
209
Category
Article
ISSN
0003-2697

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✦ Synopsis

An enzymatic method for preparing 6R-[U-14C]-tetrahydrobiopterin from [U-14C]GTP is presented. This method utilizes purified preparations of three biosynthetic enzymes for 6R-tetrahydrobiopterin, i.e., Escherichia coli GTP cyclohydrolase I, rat 6-pyruvoyl-tetrahydropterin synthase, and rat sepiapterin reductase. Based on the catalytic properties of these enzymes, the reaction conditions were optimized to complete the entire conversion reaction from GTP to 6R-tetrahydrobiopterin in a single reaction solution without the need to isolate unstable intermediates after each enzymatic reaction. The reaction conditions thereby established yielded [U-14C]biopterin in an amount equivalent to 75%, on a molar basis, of the initial amount of [U-14C]GTP. The product was subsequently isolated by high-performance liquid chromatography. The method produced labeled 6R-tetrahydrobiopterin with a specific activity of 450 Ci/mol and an overall yield of more than 60%.

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