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Structural analysis and biosynthetic engineering of a solubility-improved and less-hemolytic nystatin-like polyene in Pseudonocardia autotrophica

  • Applied genetics and molecular biotechnology
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Abstract

Polyene antibiotics such as nystatin are a large family of very valuable antifungal polyketide compounds typically produced by soil actinomycetes. Previously, using a polyene cytochrome P450 hydroxylase-specific genome screening strategy, Pseudonocardia autotrophica KCTC9441 was determined to contain an approximately 125.7-kb region of contiguous DNA with a total of 23 open reading frames, which are involved in the biosynthesis and regulation of a structurally unique polyene natural product named NPP. Here, we report the complete structure of NPP, which contains an aglycone identical to nystatin and harbors a unique di-sugar moiety, mycosaminyl-(α1-4)-N-acetyl-glucosamine. A mutant generated by inactivation of a sole glycosyltransferase gene (nppDI) within the npp gene cluster can be complemented in trans either by nppDI-encoded protein or by its nystatin counterpart, NysDI, suggesting that the two sugars might be attached by two different glycosyltransferases. Compared with nystatin (which bears a single sugar moiety), the di-sugar containing NPP exhibits approximately 300-fold higher water solubility and 10-fold reduced hemolytic activity, while retaining about 50% antifungal activity against Candida albicans. These characteristics reveal NPP as a promising candidate for further development into a pharmacokinetically improved, less-cytotoxic polyene antifungal antibiotic.

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Acknowledgements

This work was supported by the 21C Frontier Microbial Genomics and Applications Center Program, Ministry of Education, Science and Technology, Republic of Korea and the National Research Foundation of Korea (NRF) Grant (NRF-2010-616-D00030). This work was also supported in part by the grants from the 973 programs of Ministry of Science and Technology and the National Natural Science Foundation of China (D.K., L.B., Z.D., and S.L.), as well as NIH grant R01 GM076477 and the Hans W. Vahlteich Professorship (D.H.S.).

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Authors and Affiliations

  1. Department of Biological Engineering, Inha University, Incheon, 402-751, South Korea

    Mi-Jin Lee & Eung-Soo Kim

  2. State Key Laboratory of Microbial Metabolism and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200030, China

    Dekun Kong, Linquan Bai, Zixin Deng & Shuangjun Lin

  3. CHA BIO & DIOSTECH Bio Pharmaceutical Research Institute, Gangnam-gu, Seoul, 135-907, South Korea

    Kyuboem Han

  4. Life Sciences Institute and Department of Medicinal Chemistry, University of Michigan, Ann Arbor, MI, 48109-2216, USA

    David H. Sherman

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  1. Mi-Jin Lee
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Correspondence to Eung-Soo Kim.

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Mi-Jin Lee and Dekun Kong equally contributed to this paper.

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Lee, MJ., Kong, D., Han, K. et al. Structural analysis and biosynthetic engineering of a solubility-improved and less-hemolytic nystatin-like polyene in Pseudonocardia autotrophica . Appl Microbiol Biotechnol 95, 157–168 (2012). https://doi.org/10.1007/s00253-012-3955-x

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  • DOI: https://doi.org/10.1007/s00253-012-3955-x

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