Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals, College of Pharmaceutical Sciences , Zhejiang University of Technology , Hangzhou 310014 , Zhejiang , China.
State Key Laboratory of Drug Research , Shanghai Institute of Materia Medica, Chinese Academy of Sciences , Shanghai 201203 , China.
ACS Chem Biol. 2018 May 18;13(5):1200-1208. doi: 10.1021/acschembio.7b01068. Epub 2018 Apr 25.
The effect of regulatory system on the engineered biosynthetic pathway in chassis cells remains incompletely understood in microorganisms. Acyl-CoAs function as key precursors for the biosynthesis of various natural products and the dominant donors for protein acylation. The polyphenol pinosylvin, with high antimicrobial and antifungal activities, is biosynthesized with malonyl-CoA as its direct precursors. But correlation between lysine malonylation and pinosylvin biosynthesis remains unknown. Herein, we found that the malonyl-CoA-driven lysine malonylation plays an important role in interaction between the engineered pathway of pinosylvin synthesis and E. coli chassis cell. Oversupply of malonyl-CoA leads to an increase in malonylation level of global proteome as well as the enzymes in the artificial pathway, thereby decreasing yield of pinosylvin. The results revealed that the intricate balance of cellular acyl-CoA concentrations is critical for the yields of acyl-CoA-derived natural products. We next modified the enzymes in the biosynthetic pathway to adjust their acylation level and successfully improved the yield of pinosylvin. Our study uncovers the effect of protein acylation on the biosynthetic pathway, helps optimization of synthetic constructs, and provides new strategies in metabolic engineering and synthetic biology at the protein post-translational level.
在微生物中，调控系统对底盘细胞中工程化生物合成途径的影响仍不完全清楚。酰基辅酶 A 作为各种天然产物生物合成的关键前体和蛋白质酰化的主要供体发挥作用。具有高抗菌和抗真菌活性的多酚松柏醇是由丙二酰辅酶 A 作为其直接前体生物合成的。但是赖氨酸丙二酰化与松柏醇生物合成之间的相关性尚不清楚。在此，我们发现，丙二酰辅酶 A 驱动的赖氨酸丙二酰化在松柏醇合成的工程途径与大肠杆菌底盘细胞之间的相互作用中起着重要作用。丙二酰辅酶 A 的过量供应会导致全局蛋白质组以及人工途径中的酶的丙二酰化水平增加，从而降低松柏醇的产量。结果表明，细胞酰基辅酶 A 浓度的复杂平衡对酰基辅酶 A 衍生天然产物的产量至关重要。我们接下来修改生物合成途径中的酶以调整它们的酰化水平，并成功提高了松柏醇的产量。我们的研究揭示了蛋白质酰化对生物合成途径的影响，有助于优化合成构建，并在蛋白质翻译后水平的代谢工程和合成生物学中提供了新的策略。
