β-Hydroxylation of α-amino-β-hydroxylbutanoyl-glycyluridine catalyzed by a nonheme hydroxylase ensures the maturation of caprazamycin

Saeid Malek Zadeh; Mei-Hua Chen; Zhe-Chong Wang; Elahe K Astani; I-Wen Lo; Kuan-Hung Lin; Ning-Shian Hsu; Kamal Adhikari; Syue-Yi Lyu; Hsin-Ying Tsai; Yuma Terasawa; Miyuki Yabe; Kazuki Yamamoto; Satoshi Ichikawa; Tsung-Lin Li

doi:10.1038/s42004-022-00703-6

β-Hydroxylation of α-amino-β-hydroxylbutanoyl-glycyluridine catalyzed by a nonheme hydroxylase ensures the maturation of caprazamycin

Commun Chem. 2022 Jul 28;5(1):87. doi: 10.1038/s42004-022-00703-6.

Authors

Saeid Malek Zadeh^{1

2

3}, Mei-Hua Chen^{1

2

4}, Zhe-Chong Wang¹, Elahe K Astani⁵, I-Wen Lo¹, Kuan-Hung Lin¹, Ning-Shian Hsu¹, Kamal Adhikari^{1

6}, Syue-Yi Lyu¹, Hsin-Ying Tsai¹, Yuma Terasawa⁷, Miyuki Yabe⁷, Kazuki Yamamoto⁷, Satoshi Ichikawa^{7

8}, Tsung-Lin Li^{9

10

11

12}

Affiliations

¹ Genomics Research Center, Academia Sinica, Taipei, Taiwan.
² Chemical Biology and Molecular Biophysics Program, Taiwan International Graduate Program, Academia Sinica, Taipei, Taiwan.
³ Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu, Taiwan.
⁴ Department of Chemistry, National Taiwan University, Taipei, Taiwan.
⁵ Department of Chemistry, Faculty of Science, Tarbiat Modares University, Tehran, Iran.
⁶ Molecular and Biological Agricultural Sciences Program, Taiwan International Graduate Program, Academia Sinica and National Chung Hsing University, Taipei, Taiwan.
⁷ Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
⁸ Global Station for Biosurfaces and Drug Discovery, Global Institution for Collaborative Research and Education (GI- CoRE), Hokkaido University, Sapporo, Japan.
⁹ Genomics Research Center, Academia Sinica, Taipei, Taiwan. tlli@gate.sinica.edu.tw.
¹⁰ Chemical Biology and Molecular Biophysics Program, Taiwan International Graduate Program, Academia Sinica, Taipei, Taiwan. tlli@gate.sinica.edu.tw.
¹¹ Department of Chemistry, Faculty of Science, Tarbiat Modares University, Tehran, Iran. tlli@gate.sinica.edu.tw.
¹² Biotechnology Center, National Chung Hsing University, Taichung City, Taiwan. tlli@gate.sinica.edu.tw.

Abstract

Caprazamycin is a nucleoside antibiotic that inhibits phospho-N-acetylmuramyl-pentapeptide translocase (MraY). The biosynthesis of nucleoside antibiotics has been studied but is still far from completion. The present study characterized enzymes Cpz10, Cpz15, Cpz27, Mur17, Mur23 out of caprazamycin/muraymycin biosynthetic gene cluster, particularly the nonheme αKG-dependent enzyme Cpz10. Cpz15 is a β-hydroxylase converting uridine mono-phosphate to uridine 5' aldehyde, then incorporating with threonine by Mur17 (Cpz14) to form 5'-C-glycyluridine. Cpz10 hydroxylates synthetic 11 to 12 in vitro. Major product 13 derived from mutant Δcpz10 is phosphorylated by Cpz27. β-Hydroxylation of 11 by Cpz10 permits the maturation of caprazamycin, but decarboxylation of 11 by Mur23 oriented to muraymycin formation. Cpz10 recruits two iron atoms to activate dioxygen with regio-/stereo-specificity and commit electron/charge transfer, respectively. The chemo-physical interrogations should greatly advance our understanding of caprazamycin biosynthesis, which is conducive to pathway/protein engineering for developing more effective nucleoside antibiotics.

Abstract

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