The Ehrlich pathway is a major route for the renewable production of higher alcohols. However, the product scope of the Ehrlich pathway is restricted, and the product selectivity is suboptimal. Here, we demonstrate that a Coenzyme A (CoA) detour, which involves conversion of the 2-keto acids into acyl-CoAs, expands the biological toolkit of reaction chemistries available in the Ehrlich pathway to include the gamut of CoA-dependent enzymes. As a proof-of-concept, we demonstrated the first biosynthesis of a tertiary branched-alcohol, pivalcohol, at a level of ∼10 mg/L from glucose in Escherichia coli, using a pivalyl-CoA mutase from Xanthobacter autotrophicus. Furthermore, engineering an enzyme in the CoA detour, the Lactobacillus brevis CoA-acylating aldehyde dehydrogenase, allowed stringent product selectivity. Targeted production of 3-methyl-1-butanol (3-MB) in E. coli mediated by the CoA detour showed a 3-MB:side-product (isobutanol) ratio of >20, an increase over the ratios previously achieved using the conventional Ehrlich pathway.

中文翻译：

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设计辅酶绕道以扩大产品范围并增强Ehrlich途径的选择性。

Ehrlich途径是可再生生产高级醇的主要途径。但是，Ehrlich途径的产物范围受到限制，产物选择性次优。在这里，我们证明了辅酶A（CoA）绕道，其中涉及将2-酮酸转化为酰基CoA，从而扩大了Ehrlich途径中可用的反应化学生物学工具包，从而包括了CoA依赖性酶的色域。作为概念验证，我们证明了使用自生黄腐杆菌的新戊酰辅酶A变位酶，从大肠杆菌中的葡萄糖中以约10 mg / L的水平首次合成了叔支链叔醇（新戊醇）。此外，在CoA绕道中设计一种酶，即短乳杆菌CoA酰化醛脱氢酶，可以实现严格的产物选择性。