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Expanding Upon Styrene Biosynthesis to Engineer a Novel Route to 2‐Phenylethanol

22:39 EDT 11 Aug 2017 | Wiley Biotechnology Journal

2‐Phenylethanol (2PE) is a key molecule used in the fragrance and food industries, as well as a potential biofuel. In contrast to its extraction from plant biomass and/or more common chemical synthesis, microbial 2PE production has been demonstrated via both native and heterologous expression of the yeast Ehrlich pathway. Here, a novel alternative to this established pathway was systematically engineered in Escherichia coli and evaluated as a more robust and efficient route. This novel pathway was constructed via the modular extension of a previously‐engineered styrene biosynthesis pathway, proceeding from endogenous L‐phenylalanine in five steps and involving four heterologous enzymes. This 'styrene‐derived' pathway boasts a ∼10‐fold greater thermodynamic driving force than the Ehrlich pathway, and enables reduced accumulation of acetate byproduct. When directly compared using a host strain engineered for L‐phenylalanine over‐production, preservation of phosphoenolpyruvate, and reduced formation of byproduct 2‐phenylacetic acid, final 2PE titers via the styrene‐derived and Ehrlich pathways reached 1817 and 1164 mg/L, respectively, at yields of 60.6 and 38.8 mg/g. Following optimization of induction timing and initial glucose loading, 2PE titers by the styrene‐derived pathway approached as high as 2 g/L – a ∼2‐fold increase over prior reports for 2PE production by E. coli employing the Ehrlich pathway.

Original Article: Expanding Upon Styrene Biosynthesis to Engineer a Novel Route to 2‐Phenylethanol

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