@article{musiol-kroll_zubeil_schafhauser_härtner_kulik_mcarthur_koryakina_wohlleben_grond_williams_et al._2017, title={Polyketide Bioderivatization Using the Promiscuous Acyltransferase KirCII}, volume={6}, ISSN={2161-5063 2161-5063}, url={http://dx.doi.org/10.1021/acssynbio.6b00341}, DOI={10.1021/acssynbio.6b00341}, abstractNote={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.}, number={3}, journal={ACS Synthetic Biology}, publisher={American Chemical Society (ACS)}, author={Musiol-Kroll, Ewa M. and Zubeil, Florian and Schafhauser, Thomas and Härtner, Thomas and Kulik, Andreas and McArthur, John and Koryakina, Irina and Wohlleben, Wolfgang and Grond, Stephanie and Williams, Gavin J. and et al.}, year={2017}, month={Feb}, pages={421–427} }
 @article{koryakina_mcarthur_draelos_williams_2013, title={Promiscuity of a modular polyketide synthase towards natural and non-natural extender units}, volume={11}, ISSN={1477-0520 1477-0539}, url={http://dx.doi.org/10.1039/c3ob40633d}, DOI={10.1039/c3ob40633d}, abstractNote={Combinatorial biosynthesis approaches that involve modular type I polyketide synthases (PKSs) are proven strategies for the synthesis of polyketides. In general however, such strategies are usually limited in scope and utility due to the restricted substrate specificity of polyketide biosynthetic machinery. Herein, a panel of chemo-enzymatically synthesized acyl-CoA's was used to probe the promiscuity of a polyketide synthase. Promiscuity determinants were dissected, revealing that the KS is remarkably tolerant to a diverse array of extender units, while the AT likely discriminates between extender units that are native to the producing organism. Our data provides a clear blueprint for future enzyme engineering efforts, and sets the stage for harnessing extender unit promiscuity by employing various in vivo polyketide diversification strategies.}, number={27}, journal={Organic & Biomolecular Chemistry}, publisher={Royal Society of Chemistry (RSC)}, author={Koryakina, Irina and McArthur, John B. and Draelos, Matthew M. and Williams, Gavin J.}, year={2013}, pages={4449} }
 @article{koryakina_mcarthur_randall_draelos_musiol_muddiman_weber_williams_2012, title={Poly Specific trans-Acyltransferase Machinery Revealed via Engineered Acyl-CoA Synthetases}, volume={8}, ISSN={1554-8929 1554-8937}, url={http://dx.doi.org/10.1021/cb3003489}, DOI={10.1021/cb3003489}, abstractNote={Polyketide synthases construct polyketides with diverse structures and biological activities via the condensation of extender units and acyl thioesters. Although a growing body of evidence suggests that polyketide synthases might be tolerant to non-natural extender units, in vitro and in vivo studies aimed at probing and utilizing polyketide synthase specificity are severely limited to only a small number of extender units, owing to the lack of synthetic routes to a broad variety of acyl-CoA extender units. Here, we report the construction of promiscuous malonyl-CoA synthetase variants that can be used to synthesize a broad range of malonyl-CoA extender units substituted at the C2-position, several of which contain handles for chemoselective ligation and are not found in natural biosynthetic systems. We highlighted utility of these enzymes by probing the acyl-CoA specificity of several trans-acyltransferases, leading to the unprecedented discovery of poly specificity toward non-natural extender units, several of which are not found in naturally occurring biosynthetic pathways. These results reveal that polyketide biosynthetic machinery might be more tolerant to non-natural substrates than previously established, and that mutant synthetases are valuable tools for probing the specificity of biosynthetic machinery. Our data suggest new synthetic biology strategies for harnessing this promiscuity and enabling the regioselective modification of polyketides.}, number={1}, journal={ACS Chemical Biology}, publisher={American Chemical Society (ACS)}, author={Koryakina, Irina and McArthur, John and Randall, Shan and Draelos, Matthew M. and Musiol, Ewa M. and Muddiman, David C. and Weber, Tilmann and Williams, Gavin J.}, year={2012}, month={Oct}, pages={200–208} }