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Functional Characterization of NgnL, an Alpha/beta-hydrolase Enzyme Involved in Biosynthesis of Acetylated Nodusmicin

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  • Applied Microbiology
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Abstract

Nodusmicin and 18-O-acetyl-nodusmicin were isolated as a co-metabolite of nargenicin from Nocardia sp. CS682. NgnL belonging to the α/β-hydrolase superfamily was heterologously expressed in Escherichia coli BL21 and was evaluated for its potential for acetylation of nodusmicin by in-vitro reactions. The generation of 18-O-acetyl nodusmicin was confirmed by high resolution-quadruple time of flight mass spectrometry (HR-QTOF-MS). Thus, it was concluded that NgnL can perform regiospecific acetylation of nodusmicin at 18-OH in presence of acetyl-CoA as an acetate donor. The enzyme kinetic parameters as Km and Vmax of the enzymes were also evaluated. Further, the biosynthetic role of the enzyme for acetylation of nodusmicin was confirmed by gene inactivation and complementation studies. Hence, from this study, it was established that NgnL is regiospecific acetyltransferase involved in the biosynthesis of 18-O-acetyl nodusmicin in Nocardia sp. CS682.

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Acknowledgements

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (NRF-2017R1A2A2A05000939, JKS).

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  1. These authors are equally contributed.

Authors and Affiliations

  1. Institute of Biomolecule Reconstruction (TBR), Department of Life Science and Biochemical Engineering, SunMoon University, Asan, 31460, Korea

    Vijay Rayamajhi, Dipesh Dhakal & Jae Kyung Sohng

  2. Department of BT-Convergent Pharmaceutical Engineering, SunMoon University, Asan, 31460, Korea

    Jae Kyung Sohng

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  1. Vijay Rayamajhi
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  2. Dipesh Dhakal
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Correspondence to Jae Kyung Sohng.

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Rayamajhi, V., Dhakal, D. & Sohng, J.K. Functional Characterization of NgnL, an Alpha/beta-hydrolase Enzyme Involved in Biosynthesis of Acetylated Nodusmicin. Biotechnol Bioproc E 25, 414–420 (2020). https://doi.org/10.1007/s12257-019-0455-1

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