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Identification of the genes encoding enzymes involved in the early biosynthetic pathway of pteridines in Synechocystis sp. PCC 6803
Authors:Soo Woong Lee  Hee Woo Lee  Hyun Jae Chung  Young-A Kim  Yeon Jeong Kim  Yoonsoo Hahn  Jae Hoon Chung  Young Shik Park
Affiliation:Department of Microbiology, Inje University, Kimhae, South Korea.
Abstract:The biosynthetic pathway for the pteridine moiety of cyanopterine, as well as tetrahydrobiopterine, has been investigated in Synechocystis sp. PCC 6803. Open reading frames slr0426, slr1626, slr0078 and sll0330 of the organism putatively encoding GTP cyclohydrolase I, dihydroneopterine aldolase, 6-pyruvoyltetrahydropterine synthase and sepiapterine reductase, respectively, have been cloned into T7-based vectors for expression in Escherichia coli. The recombinant proteins have been purified to homogeneity and demonstrated to possess expected genuine activities except that of sll0330. Our result is the first direct evidence for the functional assignment of the open reading frames in Synechocystis sp. PCC 6803. Furthermore, the 6-pyruvoyltetrahydropterine synthase gene is demonstrated for the first time in prokaryotes. Based on the result, biosynthesis of cyanopterine is discussed.
Keywords:Synechocystis sp. PCC 6803    GTP cyclohydrolase I    6-Pyruvoyltetrahydropterine synthase    Dihydroneopterine aldolase    Heterologous overexpression    Cyanopterine
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