Key Laboratory of Combinatorial Biosynthesis and Drug Discovery Ministry of Education, School of Pharmaceutical Sciences, Wuhan University, 1 Luojiashan Rd., Wuhan, 430071, China.
State Key Laboratory of Microbial Metabolism and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Rd., Shanghai, 200240, China.
Angew Chem Int Ed Engl. 2022 Oct 24;61(43):e202208734. doi: 10.1002/anie.202208734. Epub 2022 Sep 29.
Acyl-CoAs are key precursors of primary and secondary metabolism. Their efficient biosynthesis is often impeded by the limited substrate specificity and low in vivo activity of acyl-CoA synthetases (ACSs) due to regulatory acylation of the catalytically important lysine residue in motif A10 (Lys-A10). In this study, we identified an unusual ACS (UkaQ) from the UK-2A biosynthetic pathway that naturally lacks the Lys-A10 residue and exhibits extraordinarily broad substrate specificity. Protein engineering significantly improved its stability and catalytic activity, enabling it to synthesize a large variety of acyl-CoAs with highly robust activity. By combining it with permissive carboxylases, we produced a large array of polyketide extender units and obtained six novel halobenzyl-containing antimycin analogues through an engineered biosynthetic pathway. This study significantly expands the catalytic mode of ACSs and provides a potent tool for the biosynthesis of acyl-CoA-derived natural products.
酰基辅酶 A 是初级和次级代谢的关键前体。由于酰基辅酶 A 合成酶 (ACS) 中 motif A10 (Lys-A10) 上催化重要赖氨酸残基的调节酰化，其有限的底物特异性和体内活性往往会阻碍酰基辅酶 A 的有效生物合成。在这项研究中，我们从 UK-2A 生物合成途径中鉴定出一种不寻常的 ACS (UkaQ)，它天然缺乏 Lys-A10 残基，表现出非常广泛的底物特异性。蛋白质工程显著提高了它的稳定性和催化活性，使其能够合成具有高度稳健活性的各种酰基辅酶 A。通过与允许的羧化酶结合，我们生产了大量的聚酮延伸单元，并通过工程化的生物合成途径获得了六种新型的含卤苄基安密霉素类似物。这项研究显著扩展了 ACS 的催化模式，并为酰基辅酶 A 衍生天然产物的生物合成提供了有力工具。
