Engineering of farnesyl pyrophosphate hydrolase for farnesol production in Serratia marcescens

Abstract

Farnesol (FOH), a prized sesquiterpenoid alcohol, is at the core of this study, which outlines a synthetic biology strategy to significantly boost its production for use in flavors, fragrances, pharmaceuticals, and biofuels. We constructed an efficient FOH biosynthetic pathway in Serratia marcescens, leveraging rational engineering strategies to optimize its production. Initially, we introduced a heterologous mevalonate (MVA) pathway into S. marcescens for FOH biosynthesis. We then screened different sources of monophosphate phosphatases and performed rational modifications to enhance their activity. Computational simulations were employed to model the SmAp-FP complex, guiding protein engineering efforts. The engineered strain S. marcescens SPF6_L2 achieved a FOH titer of 457.3 ± 23.1 mg/L in shake flask fermentation, which was further scaled up to 1784.3 mg/L in a 5 L fermenter. This represents one of the highest reported titers of FOH production in microorganisms to date. Our approach integrates genetic engineering, enzyme optimization, and bioprocess design to efficiently biosynthesize FOH. It sets the stage for future research on optimizing S. marcescens metabolic pathways for enhanced terpenoid biosynthesis.

Keywords

Farnesol / Phosphatases / Protein engineering / Serratia marcescens

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Jiangnan University