Diversity oriented biosynthesis via accelerated evolution of modular gene clusters.

Authors: Wlodek, Aleksandra and Kendrew, Steve G and Coates, Nigel J and Hold, Adam and Pogwizd, Joanna and Rudder, Steven and Sheehan, Lesley S and Higginbotham, Sarah J and Stanley-Smith, Anna E and Warneck, Tony and Nur-E-Alam, Mohammad and Radzom, Markus and Martin, Christine J and Overvoorde, Lois and Samborskyy, Markiyan and Alt, Silke and Heine, Daniel and Carter, Guy T and Graziani, Edmund I and Koehn, Frank E and McDonald, Leonard and Alanine, Alexander and Rodríguez Sarmiento, Rosa María and Chao, Suzan Keen and Ratni, Hasane and Steward, Lucinda and Norville, Isobel H and Sarkar-Tyson, Mitali and Moss, Steven J and Leadlay, Peter F and Wilkinson, Barrie and Gregory, Matthew A

Erythromycin, avermectin and rapamycin are clinically useful polyketide natural products produced on modular polyketide synthase multienzymes by an assembly-line process in which each module of enzymes in turn specifies attachment of a particular chemical unit. Although polyketide synthase encoding genes have been successfully engineered to produce novel analogues, the process can be relatively slow, inefficient, and frequently low-yielding. We now describe a method for rapidly recombining polyketide synthase gene clusters to replace, add or remove modules that, with high frequency, generates diverse and highly productive assembly lines. The method is exemplified in the rapamycin biosynthetic gene cluster where, in a single experiment, multiple strains were isolated producing new members of a rapamycin-related family of polyketides. The process mimics, but significantly accelerates, a plausible mechanism of natural evolution for modular polyketide synthases. Detailed sequence analysis of the recombinant genes provides unique insight into the design principles for constructing useful synthetic assembly-line multienzymes.

Journal: Nature communications
DOI: 10.1038/s41467-017-01344-3
Year: 2017

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