Acetone production with metabolically engineered strains of Acetobacterium woodii
Abstract
Expected depletion of oil and fossil resources urges the development of new alternative routes for the production of bulk chemicals and fuels beyond petroleum resources. In this study, the clostridial acetone pathway was used for the formation of acetone in the acetogenic bacterium Acetobacterium woodii. The acetone production operon (APO) containing the genes thlA (encoding thiolase A), ctfA/ctfB (encoding CoA transferase), and adc (encoding acetoacetate decarboxylase) from Clostridium acetobutylicum were cloned under the control of the thlA promoter into four vectors having different replicons for Gram-positives (pIP404, pBP1, pCB102, and pCD6). Stable replication was observed for all constructs. A. woodii [pJIR_actthlA] achieved the maximal acetone concentration under autotrophic conditions (15.2±3.4mM). Promoter sequences of the genes ackA from A. woodii and pta-ack from C. ljungdahlii were determined by primer extension (PEX) and cloned upstream of the APO. The highest acetone production in recombinant A. woodii cells was achieved using the promoters PthlA and Ppta-ack. Batch fermentations using A. woodii [pMTL84151_actthlA] in a bioreactor revealed that acetate concentration had an effect on the acetone production, due to the high Km value of the CoA transferase. In order to establish consistent acetate concentration within the bioreactor and to increase biomass, a continuous fermentation process for A. woodii was developed. Thus, acetone productivity of the strain A. woodii [pMTL84151_actthlA] was increased from 1.2mgL(-1)h(-1) in bottle fermentation to 26.4mgL(-1)h(-1) in continuous gas fermentation.
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A. woodii with plasmid-based expression of C. acetobutylicum acetone biosynthetic genes produced 26.4 mg/L/h acetone continuously from synthetic syngas in a CSTR.
“| A. woodii | Plasmid based expression of C. acetobutylicum acetone biosynthetic genes (thlA-ctfAB-adc) using different combinations of promoter (ackA, pta-ack) and plasmid origin (pIP404, pBP1, pCB102, and pCD6) | Continuous production of 26.4 mg/L/h acetone from synthetic syngas in CSTR | Hoffmeister et al., 2016 |”
From Gas Fermentation—A Flexible Platform for Commercial Scale Production of Low-Carbon-Fuels and Chemicals from Waste and Renewable Feedstocks (2016) · cites this paper as Hoffmeister et al., 2016
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- Authors
- Hoffmeister, S., Gerdom, M., Bengelsdorf, F. R., Linder, S., Flüchter, S., Öztürk, H., Blümke, W., May, A., Fischer, R.-J., Bahl, H., & Dürre, P.
- Affiliations
- Universität Ulm (DE)
- Evonik (Germany) (DE)
- University of Rostock (DE)
- Corresponding author
- Hoffmeister, Sabrina
- Venue
- Metabolic Engineering
- Publisher
- Elsevier BV
- Published
- 2016-07
- Volume
- 36
- Pages
- 37-47
- ISSN
- 1096-7176 (ISSN-L)
- OpenAlex
- W2294276789
- Cited by
- 139
- References
- 42
- Citations by year
- 2026: 9 · 2025: 5 · 2024: 12 · 2023: 18 · 2022: 24 · 2021: 17 · 2020: 15 · 2019: 17 · 2018: 6 · 2017: 9 · 2016: 7
- FWCI
- 6.75
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Metabolic Engineering (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Bundesministerium für Bildung und Forschung (01RC1105A/C)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Physical Sciences › Engineering › Biomedical Engineering › Biofuel production and bioconversion
- Physical Sciences › Engineering › Building and Construction › Anaerobic Digestion and Biogas Production
- Keywords
- acetone production, Acetobacterium woodii, CoA transferase, acetoacetate decarboxylase, promoter cloning, acetogenic bacteria, Clostridium acetobutylicum
- MeSH terms
- Acetates, Acetone, Carbon Dioxide, Recombinant Proteins, Genetic Enhancement, Acetobacterium, Metabolic Networks and Pathways, Biosynthetic Pathways, Metabolic Engineering
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