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Metabolic Checkpoint Aldehyde Dehydrogenases Are Important for Diverting β-Oxidation into 1-Butanol Biosynthesis from Kitchen Waste Oil in Pseudomonas aeruginosa

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Abstract

1-Butanol (1-BD) is a promising fuel additive which can be biosynthesized via reversed β-oxidation pathway in bacteria. However, heterologous reversed β-oxidation pathway is a carbon chain prolongation process with several genes overexpressed in most of bacterial hosts, leading to low titer of 1-BD and high cost for production. Here we displayed a forward β-oxidation pathway for 1-BD production in a kitchen waste oil (KWO) degrading Pseudomonas aeruginosa PA-3, and we proved that aldehyde dehydrogenase (ALDH) is a checkpoint for diverting metabolic flux into 1-BD biosynthesis. With nitrogen source supplied, titer of 1-BD was increased accompanied with 12 ALDH coding genes transcriptionally promoted to different degrees. At the same time, binding energies of these ALDHs with different length of acyl-CoAs in β-oxidation were calculated to identify their specificities. Based on the above information, ALDH deletions were conducted. We certified that deletion of ALDH8 and ALDH9 led to significant decreased titers of 1-BD. Finally, these two ALDHs were separately overexpressed in PA-3, and titer of 1-BD was promoted to 1.36 g/L at 72 h in shake flask. Totally in this work, we provided a forward β-oxidation pathway for 1-BD production from KWO, and the roles of ALDHs were confirmed.

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Abbreviations

KWO:

Kitchen waste oil

ALDH:

Aldehyde dehydrogenase

ADHP:

Alcohol dehydrogenase

1-BD:

1-Butanol

AA:

Amino acid

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Funding

This work was financially supported by Beijing Municipal Education Commission Technology Plan (KM202011417006), Project of Beijing Municipal Commission of Education (KZ201911417049), Premium Funding Project for Academic Human Resources Development in Beijing Union University (BPHR2018BZ01), and China Scholarship Council Project (201908110070).

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Authors and Affiliations

  1. Beijing Key Laboratory of Biomass Waste Resource Utilization, College of Biochemical Engineering, Beijing Union University, Beijing, 100023, China

    Ying Li, Zheng Liu & Xizhen Ge

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  1. Ying Li
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  2. Zheng Liu
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Correspondence to Xizhen Ge.

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Li, Y., Liu, Z. & Ge, X. Metabolic Checkpoint Aldehyde Dehydrogenases Are Important for Diverting β-Oxidation into 1-Butanol Biosynthesis from Kitchen Waste Oil in Pseudomonas aeruginosa. Appl Biochem Biotechnol 193, 730–742 (2021). https://doi.org/10.1007/s12010-020-03456-x

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  • DOI: https://doi.org/10.1007/s12010-020-03456-x

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