Precursor Supply for Erythromycin Biosynthesis: Engineering of Propionate Assimilation Pathway Based on Propionylation Modification
Precursor Supply for Erythromycin Biosynthesis: Engineering of Propionate Assimilation Pathway Based on Propionylation Modification
复制标题
红霉素生物合成的前体供应：基于丙酰化修饰的丙酸同化途径工程
DOI：
10.1021/acssynbio.8b00396
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发表时间：
2019
影响因子：
4.7
通讯作者：
Ye Bang Ce
中科院分区：
文献类型：
--
作者：
You Di;Wang Miao Miao;Yin Bin Cheng;Ye Bang Ce
红霉素在医学治疗中是必需的，并且已知是用丙酰辅酶A作为直接前体生物合成的。丙酰辅酶A的过量供应会诱导过度丙酰化，这被证明对红糖多孢菌中的红霉素合成有害。在此，我们鉴定了三种受丙酰化调节的丙酰辅酶A合成酶，并且一种丙酰辅酶A合成酶SACE_1780显示出对丙酰化的抗性。通过两种方法开发了一种绕过丙酰化引起的反馈抑制来提高前体（丙酰辅酶A）供应的实用策略：丙酰转移酶 AcuA 的删除和 SACE_1780 的过表达。构建的 ΔacuAstrain 红霉素产量提高了 10%； SACE_1780过表达菌株产生的红霉素产量比野生型菌株NRRL2338高33%，比工业高产Ab菌株高22%。这些发现揭示了蛋白质酰化在抗生素生物合成前体供应中的作用，并为合成生物学中改善次级代谢产物提供了有效的翻译后修饰代谢工程策略（称为PTM-ME）。
Erythromycin is necessary in medical treatment and known to be biosynthesized with propionyl-CoA as direct precursor. Oversupply of propionyl-CoA induced hyperpropionylation, which was demonstrated as harmful for erythromycin synthesis inSaccharopolyspora erythraea. Herein, we identified three propionyl-CoA synthetases regulated by propionylation, and one propionyl-CoA synthetase SACE_1780 revealed resistance to propionylation. A practical strategy for raising the precursor (propionyl-CoA) supply bypassing the feedback inhibition caused by propionylation was developed through two approaches: deletion of the propionyltransferase AcuA, and SACE_1780 overexpression. The constructed ΔacuAstrain presented a 10% increase in erythromycin yield; SACE_1780 overexpression strain produced 33% higher erythromycin yield than the wildtype strain NRRL2338 and 22% higher erythromycin yield than the industrial high yield Ab strain. These findings uncover the role of protein acylation in precursor supply for antibiotics biosynthesis and provide efficient post-translational modification-metabolic engineering strategy (named as PTM-ME) in synthetic biology for improvement of secondary metabolites.