Overexpression of key enzymes of the 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway for improving squalene production in Escherichia coli

Liu, Haiyuan; Han, Song; Xie, Liping; Pan, Jie; Zhang, Wei; Gong, Guihua; Youjia, HU

doi:10.5897/ajb2017.16235

Cited by 6 publications

(1 citation statement)

References 23 publications

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“…The identification and quantification of squalene was conducted according to a reported study. 21 Each strain was harvested from a 20 mL culture medium and added into a 50 mL centrifugal cube. The suspension was centrifuged at 8,000 rpm for 5 minutes.…”

Section: Identification and Quantification Of Squalenementioning

confidence: 99%

Squalene Biosynthesis in Engineered Escherichia Coli

Zhang,

Li,

Xie

et al. 2023

Pharmaceutical Fronts

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Squalene is a natural triterpenoid that is important in medicine, and daily-use chemical and nutraceutical industries. The demand for squalene remains high and constant. Shark liver oil is the source of the compound; however, the source is unsustainable due to the protection of the gradually extinct animal. This study aimed to construct an engineered Escherichia coli strain to produce squalene in a green and sustainable manner. In this study, the activity of several squalene synthases from different species was screened. Gas chromatography-mass spectrometry and high-performance liquid chromatography were used for chemical structure identification. The mevalonate pathway (MVA) was constructed into BL21(DE3) to enhance the supply of farnesyl diphosphate to obtain squalene at a concentration of 69.3 mg/L. IspH was overexpressed in BL21(DE3) to decrease squalene production due to the accumulation of dimethylallyl diphosphate. By balancing the overexpression of ispH and ispG, the production of squalene increased to 298.3 mg/L. The genes of the MVA pathway were further separated upstream and downstream, and constructed into two plasmids with different origins to balance the metabolic flux. By optimizing the expression strength of genes in MVA and 2-C-methyl-d-erythritol-4-phosphate pathway, an engineered strain was finally obtained with squalene production of 974.3 mg/L. By replacing chassis cell BL21(DE3) with XL1-Blue, squalene production was further increased to 1,274 mg/L.

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Section: Identification and Quantification Of Squalenementioning

confidence: 99%

Squalene Biosynthesis in Engineered Escherichia Coli

Zhang,

Li,

Xie

et al. 2023

Pharmaceutical Fronts

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Metabolic Engineering for the Biosynthesis of Terpenoids from Microbial Cell Factories

Shukla,

Gupta,

Phulara

2023

Microbial Bioactive Compounds

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Engineering plant family TPS into cyanobacterial host for terpenoids production

Rautela

Kumar

2022

Plant Cell Rep

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Terpenoids are synthesized naturally by plants as secondary metabolites, and are diverse and complex in structure with multiple applications in bioenergy, food, cosmetics, and medicine. This makes the production of terpenoids such as isoprene, β-phellandrene, farnesene, amorphadiene, and squalene valuable, owing to which their industrial demand cannot be fulfilled exclusively by plant sources. They are synthesized via the Methylerythritol phosphate pathway (MEP) and the Mevalonate pathway (MVA), both existing in plants. The advent of genetic engineering and the latest accomplishments in synthetic biology and metabolic engineering allow microbial synthesis of terpenoids. Cyanobacteria manifest to be the promising hosts for this, utilizing sunlight and CO 2 . Cyanobacteria possess MEP pathway to generate precursors for terpenoid synthesis. The terpenoid synthesis can be amplified by overexpressing the MEP pathway and engineering MVA pathway genes. According to the desired terpenoid, terpene synthases unique to the plant kingdom must be incorporated in cyanobacteria. Engineering an organism to be used as a cell factory comes with drawbacks such as hampered cell growth and disturbance in metabolic flux. This review set forth a comparison between MEP and MVA pathways, strategies to overexpress these pathways with their challenges.

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Overexpression of key enzymes of the 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway for improving squalene production in Escherichia coli

Cited by 6 publications

References 23 publications

Squalene Biosynthesis in Engineered Escherichia Coli

Squalene Biosynthesis in Engineered Escherichia Coli

Metabolic Engineering for the Biosynthesis of Terpenoids from Microbial Cell Factories

Engineering plant family TPS into cyanobacterial host for terpenoids production

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