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Carbon partitioning to the terpenoid biosynthetic pathway enables heterologous β-phellandrene production in Escherichia coli cultures

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Abstract

Escherichia coli was used as a microbial system for the heterologous synthesis of β-phellandrene, a monoterpene of plant origin with several potential commercial applications. Expression of Lavandula angustifolia β-phellandrene synthase (PHLS), alone or in combination with Picea abies geranyl-diphosphate synthase in E. coli, resulted in no β-phellandrene accumulation, in sharp contrast to observations with PHLS-transformed cyanobacteria. Lack of β-phellandrene biosynthesis in E. coli was attributed to the limited endogenous carbon partitioning through the native 2-C-methylerythritol-4-phosphate (MEP) pathway. Heterologous co-expression of the mevalonic acid pathway, enhancing cellular carbon partitioning and flux toward the universal isoprenoid precursors, isopentenyl-diphosphate and dimethylallyl-diphosphate, was required to confer β-phellandrene production. Differences in endogenous carbon flux toward the synthesis of isoprenoids between photosynthetic (Synechocystis) and non-photosynthetic bacteria (E. coli) are discussed in terms of differences in the regulation of carbon partitioning through the MEP biosynthetic pathway in the two systems.

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Abbreviations

DCW:

Dry cell weight

DMAPP:

Dimethylallyl-diphosphate

GPPS:

Geranyl-diphosphate synthase

IPP:

Isopentenyl-diphosphate

IPTG:

Isopropyl β-d-1-thiogalactopyranoside

MEP:

2-C-methyl-erythritol-4-phosphate

MVA:

Mevalonic acid

OD:

Optical density

β-PHL:

β-Phellandrene

PHLS:

β-Phellandrene synthase

TIR:

Translation initiation region

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Authors and Affiliations

  1. Department of Plant and Microbial Biology, University of California, 111 Koshland Hall, Berkeley, CA, 94720-3102, USA

    Cinzia Formighieri & Anastasios Melis

Authors
  1. Cinzia Formighieri
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  2. Anastasios Melis
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Correspondence to Anastasios Melis.

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Communicated by Erko Stackebrandt.

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Formighieri, C., Melis, A. Carbon partitioning to the terpenoid biosynthetic pathway enables heterologous β-phellandrene production in Escherichia coli cultures. Arch Microbiol 196, 853–861 (2014). https://doi.org/10.1007/s00203-014-1024-9

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  • DOI: https://doi.org/10.1007/s00203-014-1024-9

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