1. Gatekeeping Ketosynthases Dictate Initiation of Assembly Line Biosynthesis of Pyrrolic Polyketides
- Author
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Will R. Gutekunst, Dongqi Yi, James C. Gumbart, Vinayak Agarwal, and Atanu Acharya
- Subjects
Substrate Specificities ,Biological Products ,biology ,Chemistry ,Stereochemistry ,High selectivity ,Active site ,Rational engineering ,Context (language use) ,General Chemistry ,010402 general chemistry ,01 natural sciences ,Biochemistry ,Catalysis ,0104 chemical sciences ,Polyketide ,chemistry.chemical_compound ,Colloid and Surface Chemistry ,Biosynthesis ,Polyketides ,biology.protein ,Assembly line ,Polyketide Synthases - Abstract
Assembly line biosynthesis of polyketide natural products involves checkpoints where identities of thiotemplated intermediates are verified before polyketide extension reactions are allowed to proceed. Determining what these checkpoints are and how they operate is critical for reprogramming polyketide assembly lines. Here we demonstrate that ketosynthase (KS) domains can perform this gatekeeping role. By comparing the substrate specificities for polyketide synthases that extend pyrrolyl and halogenated pyrrolyl substrates, we find that KS domains that need to differentiate between these two substrates exercise high selectivity. We additionally find that amino acid residues in the KS active site facilitate this selectivity and that these residues are amenable to rational engineering. On the other hand, KS domains that do not need to make selectivity decisions in their native physiological context are substrate-promiscuous. We also provide evidence that delivery of substrates to polyketide synthases by non-native carrier proteins is accompanied by reduced biosynthetic efficiency.
- Published
- 2021