Chemoinformatic-guided engineering of polyketide synthases
Zargar, A.; Lal, R.; Valencia, L.; Wang, J.; Backman, T.; Cruz-Morales, P.; Kothari, A.; Werts, M.; Wong, A.; Bailey, C.; Loubat, A.; Liu, Y.; Benites, V.; Chang, S.; Hernández, A.; Barajas, J.; Thompson, M.; Barcelos, C.; Anayah, R.; Garcia Martin, H.; Mukhopadhyay, A.; Baidoo, E.; Katz, L.; Keasling, J.
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Polyketide synthase (PKS) engineering is an attractive method to generate new molecules such as commodity, fine and specialty chemicals. A central challenge in PKS design is replacing a partially reductive module with a fully reductive module through a reductive loop exchange, thereby generating a saturated β-carbon. In this work, we sought to establish an engineering strategy for reductive loop exchanges based on chemoinformatics, a field traditionally used in drug discovery. We first introduced a set of donor reductive loops of diverse genetic origin and chemical substrate structures into the first extension module of the lipomycin PKS (LipPKS1). These results demonstrated that chemical similarity between the substrate of the donor loops and recipient LipPKS1 correlated with product titers. Consequently, we identified donor loops with substrates chemically similar to LipPKS1 for further reductive loop exchanges, and we observed a statistically significant correlation with production. Reductive loops with the highest chemical similarity resulted in production of branched, short-chain fatty acids reaching a titer of 165 mg/L in Streptomyces albus J1074. Collectively, our work formulizes a new chemoinformatic paradigm for de novo PKS biosynthesis which may accelerate the production of valuable bioproducts.