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Isolation and characterization of pcsB, the gene for a polyene carboxamide synthase that tailors pimaricin into AB-400

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Abstract

From cell-free extracts of Streptomyces RGU5.3, a tailoring activity of pimaricin, leading to the biosynthesis of its natural carboxamide derivative AB-400, was recently identified. The two polyene macrolides, pimaricin and AB-400, were produced in almost equal quantities and can be detected in the fermentation broth of the producer strain. This report concerns the isolation and partial characterization of the gene, polyene carboxamide synthase (pcsB), responsible for the bioconversion. The gene encoded an asparagine synthase-like protein, belonging to the type II glutamine amidotransferase family, and was named pcsB. The fermentation broth of a recombinant strain carrying the engineered pcsB gene under the control of the inducible tipA promoter within an integrative vector produces the carboxamide AB-400 as the main polyene macrolide.

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Acknowledgments

This work was supported by Grants from Spanish Ministerio de Ciencia e Innovación BIO2005-02785, BIO2008-03683, and CIT-010000-2008-4. We do not have any conflict of interest which may affect either the results or our interpretation. We thank Sir Prof. David A. Hopwood for critical reading the manuscript. We thank C. Nieto for her excellent technical help.

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  1. Centro Nacional de Biotecnología del CSIC, Campus de la UAM, Darwin 3, Cantoblanco, 28049, Madrid, Spain

    Domingo Miranzo, Elena M. Seco, Trinidad Cuesta & Francisco Malpartida

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  1. Domingo Miranzo
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  2. Elena M. Seco
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  3. Trinidad Cuesta
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  4. Francisco Malpartida
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Correspondence to Francisco Malpartida.

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Miranzo, D., Seco, E.M., Cuesta, T. et al. Isolation and characterization of pcsB, the gene for a polyene carboxamide synthase that tailors pimaricin into AB-400. Appl Microbiol Biotechnol 85, 1809–1819 (2010). https://doi.org/10.1007/s00253-009-2195-1

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