Polyketide Bioderivatization Using the Promiscuous Acyltransferase KirCII.

During polyketide biosynthesis, acyltransferases (ATs) are the essential gatekeepers which provide the assembly lines with precursors and thus contribute greatly to structural diversity. Previously, we demonstrated that the discrete AT KirCII from the kirromycin antibiotic pathway accesses nonmalonate extender units. Here, we exploit the promiscuity of KirCII to generate new kirromycins with allyl- and propargyl-side chains in vivo, the latter were utilized as educts for further modification by "click" chemistry.

[1]  T. Weber,et al.  The AT2 Domain of KirCI Loads Malonyl Extender Units to the ACPs of the Kirromycin PKS , 2013, Chembiochem : a European journal of chemical biology.

[2]  A. Parmeggiani,et al.  Mechanism of the inhibition of protein synthesis by kirromycin. Role of elongation factor Tu and ribosomes. , 1977, European journal of biochemistry.

[3]  W. Wohlleben,et al.  The ABC transporter Tba of Amycolatopsis balhimycina is required for efficient export of the glycopeptide antibiotic balhimycin , 2007, Applied Microbiology and Biotechnology.

[4]  Gavin J. Williams,et al.  Mutant Malonyl‐CoA Synthetases with Altered Specificity for Polyketide Synthase Extender Unit Generation , 2011, Chembiochem : a European journal of chemical biology.

[5]  Tae-Wan Kim,et al.  Continuous cell-free protein synthesis using glycolytic intermediates as energy sources. , 2008, Journal of microbiology and biotechnology.

[6]  Dariusz Matosiuk,et al.  Click chemistry for drug development and diverse chemical-biology applications. , 2013, Chemical reviews.

[7]  J. Swartz,et al.  An Economical Method for Cell‐Free Protein Synthesis using Glucose and Nucleoside Monophosphates , 2008, Biotechnology progress.

[8]  G. Gallo,et al.  Mass spectrometric techniques for structure and novelty determination of kirromycin-like antibiotics , 1992 .

[9]  J. Piel,et al.  Analysis of the Sorangicin Gene Cluster Reinforces the Utility of a Combined Phylogenetic/Retrobiosynthetic Analysis for Deciphering Natural Product Assembly by trans‐AT PKS , 2010, Chembiochem : a European journal of chemical biology.

[10]  B. Shen,et al.  Oxazolomycin Biosynthesis in Streptomyces albus JA3453 Featuring an “Acyltransferase-less” Type I Polyketide Synthase That Incorporates Two Distinct Extender Units* , 2010, The Journal of Biological Chemistry.

[11]  R. Hilgenfeld,et al.  Conformational Change of Elongation Factor Tu (EF-Tu) Induced by Antibiotic Binding , 2001, The Journal of Biological Chemistry.

[12]  Frank Schulz,et al.  Predicted Incorporation of Non‐native Substrates by a Polyketide Synthase Yields Bioactive Natural Product Derivatives , 2014, Chembiochem : a European journal of chemical biology.

[13]  T. Weber,et al.  Molecular analysis of the kirromycin biosynthetic gene cluster revealed beta-alanine as precursor of the pyridone moiety. , 2008, Chemistry & biology.

[14]  Tae-Wan Kim,et al.  An economical and highly productive cell-free protein synthesis system utilizing fructose-1,6-bisphosphate as an energy source. , 2007, Journal of biotechnology.

[15]  T. Weber,et al.  Discrete acyltransferases involved in polyketide biosynthesis , 2012 .

[16]  C. Hertweck,et al.  The biosynthetic logic of polyketide diversity. , 2009, Angewandte Chemie.

[17]  Tilmann Weber,et al.  Reprogramming acyl carrier protein interactions of an Acyl-CoA promiscuous trans-acyltransferase. , 2014, Chemistry & biology.

[18]  Frank Schulz,et al.  Enzyme-directed mutasynthesis: a combined experimental and theoretical approach to substrate recognition of a polyketide synthase. , 2013, ACS chemical biology.

[19]  C. Walsh,et al.  Prospects for new antibiotics: a molecule-centered perspective , 2013, The Journal of Antibiotics.

[20]  Chaitan Khosla,et al.  Engineering the acyltransferase substrate specificity of assembly line polyketide synthases , 2013, Journal of The Royal Society Interface.

[21]  Yeo Joon Yoon,et al.  Biosynthesis of the allylmalonyl-CoA extender unit for the FK506 polyketide synthase proceeds through a dedicated polyketide synthase and facilitates the mutasynthesis of analogues. , 2011, Journal of the American Chemical Society.

[22]  Tilmann Weber,et al.  Poly specific trans-acyltransferase machinery revealed via engineered acyl-CoA synthetases. , 2013, ACS chemical biology.

[23]  T. Weber,et al.  Supramolecular templating in kirromycin biosynthesis: the acyltransferase KirCII loads ethylmalonyl-CoA extender onto a specific ACP of the trans-AT PKS. , 2011, Chemistry & biology.

[24]  Jung-Won Keum,et al.  Simple procedures for the construction of a robust and cost-effective cell-free protein synthesis system. , 2006, Journal of biotechnology.

[25]  Jörn Piel,et al.  Biosynthesis of polyketides by trans-AT polyketide synthases. , 2016, Natural product reports.

[26]  David H Sherman,et al.  Inversion of Extender Unit Selectivity in the Erythromycin Polyketide Synthase by Acyltransferase Domain Engineering. , 2017, ACS chemical biology.

[27]  R G Kim,et al.  Expression-independent consumption of substrates in cell-free expression system from Escherichia coli. , 2000, Journal of biotechnology.

[28]  A. Parmeggiani,et al.  Kirromycin, an inhibitor of protein biosynthesis that acts on elongation factor Tu. , 1974, Proceedings of the National Academy of Sciences of the United States of America.

[29]  M. Bibb,et al.  Elongation Factor Tu3 (EF-Tu3) from the Kirromycin Producer Streptomyces ramocissimus Is Resistant to Three Classes of EF-Tu-Specific Inhibitors , 2007, Journal of bacteriology.