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Endogenous CRISPR/Cas systems for genome engineering in the acetogens Acetobacterium woodii and Clostridium autoethanogenum (2023)
Journal Article
Poulalier-Delavelle, M., Baker, J. P., Millard, J., Winzer, K., & Minton, N. P. (2023). Endogenous CRISPR/Cas systems for genome engineering in the acetogens Acetobacterium woodii and Clostridium autoethanogenum. Frontiers in Bioengineering and Biotechnology, 11, Article 1213236. https://doi.org/10.3389/fbioe.2023.1213236

Acetogenic bacteria can play a major role in achieving Net Zero through their ability to convert CO2 into industrially relevant chemicals and fuels. Full exploitation of this potential will be reliant on effective metabolic engineering tools, such as... Read More about Endogenous CRISPR/Cas systems for genome engineering in the acetogens Acetobacterium woodii and Clostridium autoethanogenum.

The Lanthipeptide Synthetase-like Protein CA_C0082 Is an Effector of Agr Quorum Sensing in Clostridium acetobutylicum (2023)
Journal Article
Humphreys, J. R., Bean, Z., Twycross, J., & Winzer, K. (2023). The Lanthipeptide Synthetase-like Protein CA_C0082 Is an Effector of Agr Quorum Sensing in Clostridium acetobutylicum. Microorganisms, 11(6), Article 1460. https://doi.org/10.3390/microorganisms11061460

Lanthipeptide synthetases are present in all domains of life. They catalyze a crucial step during lanthipeptide biosynthesis by introducing thioether linkages during posttranslational peptide modification. Lanthipeptides have a wide range of function... Read More about The Lanthipeptide Synthetase-like Protein CA_C0082 Is an Effector of Agr Quorum Sensing in Clostridium acetobutylicum.

Improving carbon monoxide tolerance of Cupriavidus necator H16 through adaptive laboratory evolution (2023)
Journal Article
Wickham-Smith, C., Malys, N., & Winzer, K. (2023). Improving carbon monoxide tolerance of Cupriavidus necator H16 through adaptive laboratory evolution. Frontiers in Bioengineering and Biotechnology, 11, https://doi.org/10.3389/fbioe.2023.1178536

Background: The toxic gas carbon monoxide (CO) is abundantly present in synthesis gas (syngas) and certain industrial waste gases that can serve as feedstocks for the biological production of industrially significant chemicals and fuels. For efficien... Read More about Improving carbon monoxide tolerance of Cupriavidus necator H16 through adaptive laboratory evolution.

Growth rate and nutrient limitation as key drivers of extracellular quorum sensing signal molecule accumulation in Pseudomonas aeruginosa (2023)
Journal Article
Dubern, J., Halliday, N., Cámara, M., Winzer, K., Barrett, D. A., Hardie, K. R., & Williams, P. (2023). Growth rate and nutrient limitation as key drivers of extracellular quorum sensing signal molecule accumulation in Pseudomonas aeruginosa. Microbiology, 169(4), Article 001316. https://doi.org/10.1099/mic.0.001316

In Pseudomonas aeruginosa, quorum sensing (QS) depends on an interconnected regulatory hierarchy involving the Las, Rhl and Pqs systems, which are collectively responsible for the co-ordinated synthesis of a diverse repertoire of N-acylhomoserine lac... Read More about Growth rate and nutrient limitation as key drivers of extracellular quorum sensing signal molecule accumulation in Pseudomonas aeruginosa.

Clostridium beijerinckii strain degeneration is driven by the loss of Spo0A activity (2023)
Journal Article
Humphreys, J. R., Debebe, B. J., Diggle, S. P., & Winzer, K. (2023). Clostridium beijerinckii strain degeneration is driven by the loss of Spo0A activity. Frontiers in Microbiology, 13, Article 1075609. https://doi.org/10.3389/fmicb.2022.1075609

Solventogenic clostridia represent a diverse group of anaerobic, spore-forming bacteria capable of producing acetone, butanol and ethanol through their unique biphasic metabolism. An intrinsic problem with these organisms however is their tendency to... Read More about Clostridium beijerinckii strain degeneration is driven by the loss of Spo0A activity.

Metabolic engineering of Cupriavidus necator H16 for heterotrophic and autotrophic production of 3-hydroxypropionic acid (2022)
Journal Article
Salinas, A., McGregor, C., Irorere, V., Arenas-López, C., Bommareddy, R. R., Winzer, K., …Kovács, K. (2022). Metabolic engineering of Cupriavidus necator H16 for heterotrophic and autotrophic production of 3-hydroxypropionic acid. Metabolic Engineering, 74, 178-190. https://doi.org/10.1016/j.ymben.2022.10.014

3-Hydroxypropionate (3-HP) is a versatile compound for chemical synthesis and a potential building block for biodegradable polymers. Cupriavidus necator H16, a facultative chemolithoautotroph, is an attractive production chassis and has been extensiv... Read More about Metabolic engineering of Cupriavidus necator H16 for heterotrophic and autotrophic production of 3-hydroxypropionic acid.

Metabolic modeling-based drug repurposing in Glioblastoma (2022)
Journal Article
Tomi-Andrino, C., Pandele, A., Winzer, K., King, J., Rahman, R., & Kim, D. (2022). Metabolic modeling-based drug repurposing in Glioblastoma. Scientific Reports, 12, Article 11189. https://doi.org/10.1038/s41598-022-14721-w

The manifestation of intra- and inter-tumor heterogeneity hinders the development of ubiquitous cancer treatments, thus requiring a tailored therapy for each cancer type. Specifically, the reprogramming of cellular metabolism has been identified as a... Read More about Metabolic modeling-based drug repurposing in Glioblastoma.

A genome-scale metabolic model of Cupriavidus necator H16 integrated with TraDIS and transcriptomic data reveals metabolic insights for biotechnological applications (2022)
Journal Article
Pearcy, N., Garavaglia, M., Millat, T., Gilbert, J. P., Song, Y., Hartman, H., …Minton, N. P. (2022). A genome-scale metabolic model of Cupriavidus necator H16 integrated with TraDIS and transcriptomic data reveals metabolic insights for biotechnological applications. PLoS Computational Biology, 18(5), Article e1010106. https://doi.org/10.1371/journal.pcbi.1010106

Exploiting biological processes to recycle renewable carbon into high value platform chemicals provides a sustainable and greener alternative to current reliance on petrochemicals. In this regard Cupriavidus necator H16 represents a particularly prom... Read More about A genome-scale metabolic model of Cupriavidus necator H16 integrated with TraDIS and transcriptomic data reveals metabolic insights for biotechnological applications.

Required Gene Set for Autotrophic Growth of Clostridium autoethanogenum (2022)
Journal Article
Woods, C., Humphreys, C. M., Tomi-Andrino, C., Henstra, A. M., Köpke, M., Simpson, S. D., …Minton, N. P. (2022). Required Gene Set for Autotrophic Growth of Clostridium autoethanogenum. Applied and Environmental Microbiology, 88(7), https://doi.org/10.1128/aem.02479-21

The majority of the genes present in bacterial genomes remain poorly characterized, with up to one-third of those that are protein encoding having no definitive function. Transposon insertion sequencing represents a high-throughput technique that can... Read More about Required Gene Set for Autotrophic Growth of Clostridium autoethanogenum.

Agr Quorum Sensing influences the Wood-Ljungdahl pathway in Clostridium autoethanogenum (2022)
Journal Article
Piatek, P., Humphreys, C., Raut, M., Wright, P. C., Simpson, S., Köpke, M., …Winzer, K. (2022). Agr Quorum Sensing influences the Wood-Ljungdahl pathway in Clostridium autoethanogenum. Scientific Reports, 12(1), Article 411. https://doi.org/10.1038/s41598-021-03999-x

Acetogenic bacteria are capable of fermenting CO2 and carbon monoxide containing waste-gases into a range of platform chemicals and fuels. Despite major advances in genetic engineering and improving these biocatalysts, several important physiological... Read More about Agr Quorum Sensing influences the Wood-Ljungdahl pathway in Clostridium autoethanogenum.

Quantitative Bioreactor Monitoring of Intracellular Bacterial Metabolites in Clostridium autoethanogenum Using Liquid Chromatography–Isotope Dilution Mass Spectrometry (2021)
Journal Article
Safo, L., Abdelrazig, S., Grosse-Honebrink, A., Millat, T., Henstra, A. M., Norman, R., …Barrett, D. A. (2021). Quantitative Bioreactor Monitoring of Intracellular Bacterial Metabolites in Clostridium autoethanogenum Using Liquid Chromatography–Isotope Dilution Mass Spectrometry. ACS Omega, 6, 13518-13526. https://doi.org/10.1021/acsomega.0c05588

We report a liquid chromatography–isotope dilution mass spectrometry method for the simultaneous quantification of 131 intracellular bacterial metabolites of Clostridium autoethanogenum. A comprehensive mixture of uniformly 13C-labeled internal stand... Read More about Quantitative Bioreactor Monitoring of Intracellular Bacterial Metabolites in Clostridium autoethanogenum Using Liquid Chromatography–Isotope Dilution Mass Spectrometry.

Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol (2021)
Journal Article
Grosse-Honebrink, A., Little, G. T., Bean, Z., Heldt, D., Cornock, R. H. M., Winzer, K., …Zhang, Y. (2021). Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol. Frontiers in Bioengineering and Biotechnology, 9, Article 659895. https://doi.org/10.3389/fbioe.2021.659895

Chirally pure (R)-1,3-butanediol ((R)-1,3-BDO) is a valuable intermediate for the production of fragrances, pheromones, insecticides and antibiotics. Biotechnological production results in superior enantiomeric excess over chemical production and is... Read More about Development of Clostridium saccharoperbutylacetonicum as a Whole Cell Biocatalyst for Production of Chirally Pure (R)-1,3-Butanediol.

Physicochemical and metabolic constraints for thermodynamics-based stoichiometric modelling under mesophilic growth conditions (2021)
Journal Article
Tomi-Andrino, C., Norman, R., Millat, T., Soucaille, P., Winzer, K., Barrett, D. A., …Kim, D. (2021). Physicochemical and metabolic constraints for thermodynamics-based stoichiometric modelling under mesophilic growth conditions. PLoS Computational Biology, 17(1), Article e1007694. https://doi.org/10.1371/journal.pcbi.1007694

© 2021 Tomi-Andrino et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source a... Read More about Physicochemical and metabolic constraints for thermodynamics-based stoichiometric modelling under mesophilic growth conditions.

Assessing the impact of physicochemical parameters in the predictive capabilities of thermodynamics-based stoichiometric approaches under mesophilic and thermophilic conditions (2020)
Working Paper
Tomi-Andrino, C., Norman, R., Millat, T., Soucaille, P., Winzer, K., Barrett, D. A., …Kim, D. Assessing the impact of physicochemical parameters in the predictive capabilities of thermodynamics-based stoichiometric approaches under mesophilic and thermophilic conditions

Metabolic engineering in the post-genomic era is characterised by the development of new methods for metabolomics and fluxomics, supported by the integration of genetic engineering tools and mathematical modelling. Particularly, constraint-based stoi... Read More about Assessing the impact of physicochemical parameters in the predictive capabilities of thermodynamics-based stoichiometric approaches under mesophilic and thermophilic conditions.

A complete genome sequence of Cupriavidus necator H16 (DSM 428) (2019)
Journal Article
Little, G. T., Ehsaan, M., Arenas-López, C., Jawed, K., Winzer, K., Kovacs, K., & Minton, N. P. (2019). A complete genome sequence of Cupriavidus necator H16 (DSM 428). Microbiology Resource Announcements, 8(37), 1-2. https://doi.org/10.1128/MRA.00814-19

The hydrogen-utilizing strain Cupriavidus necator H16 (DSM 428) was sequenced using a combination of PacBio and Illumina sequencing. Annotation of this strain reveals 6,543 protein-coding genes, 263 pseudogenes, 64 tRNA genes, and 15 rRNA genes.

The carbonic anhydrase of Clostridium autoethanogenum represents a new subclass of β-carbonic anhydrases (2019)
Journal Article
Pander, B., Harris, G., Scott, D. J., Winzer, K., Köpke, M., Simpson, S. D., …Henstra, A. M. (2019). The carbonic anhydrase of Clostridium autoethanogenum represents a new subclass of β-carbonic anhydrases. Applied Microbiology and Biotechnology, 103(17), 7275-7286. https://doi.org/10.1007/s00253-019-10015-w

Carbonic anhydrase catalyses the interconversion of carbon dioxide and water to bicarbonate and protons. It was unknown if the industrial relevant acetogen Clostridium autoethanogenum possesses these enzymes. We identified two putative carbonic anhyd... Read More about The carbonic anhydrase of Clostridium autoethanogenum represents a new subclass of β-carbonic anhydrases.

A novel conjugal donor strain for improved DNA transfer into Clostridium spp. (2019)
Journal Article
Woods, C., Humphreys, C. M., Rodrigues, R. M., Ingle, P., Rowe, P., Henstra, A. M., …Minton, N. P. (2019). A novel conjugal donor strain for improved DNA transfer into Clostridium spp. Anaerobe, 59, 184-191. https://doi.org/10.1016/j.anaerobe.2019.06.020

© 2019 The Authors Importance: The ability to transfer genetic material into a target organism is crucial for the development of a wide range of targeted genetic manipulation techniques. Overcoming the organisms’ native restriction systems which targ... Read More about A novel conjugal donor strain for improved DNA transfer into Clostridium spp..

The genetic basis of 3-hydroxypropanoate metabolism in Cupriavidus necator H16 (2019)
Journal Article
Arenas-López, C., Locker, J., Orol, D., Walter, F., Busche, T., Kalinowski, J., …Winzer, K. (2019). The genetic basis of 3-hydroxypropanoate metabolism in Cupriavidus necator H16. Biotechnology for Biofuels, 12, Article 150. https://doi.org/10.1186/s13068-019-1489-5

Background 3-Hydroxypropionic acid (3-HP) is a promising platform chemical with various industrial applications. Several metabolic routes to produce 3-HP from organic substrates such as sugars or glycerol have been implemented in yeast, enterobacter... Read More about The genetic basis of 3-hydroxypropanoate metabolism in Cupriavidus necator H16.

Genome-scale model of C. autoethanogenum reveals optimal bioprocess conditions for high-value chemical production from carbon monoxide (2019)
Journal Article
Norman, R. O., Millat, T., Schatschneider, S., Henstra, A. M., Breitkopf, R., Pander, B., …Hodgman, C. (2019). Genome-scale model of C. autoethanogenum reveals optimal bioprocess conditions for high-value chemical production from carbon monoxide. Engineering Biology, 3(2), 32-40. https://doi.org/10.1049/enb.2018.5003

Clostridium autoethanogenum is an industrial microbe used for the commercial-scale production of ethanol from carbon monoxide. While significant progress has been made in the attempted diversification of this bioprocess, further improvements are desi... Read More about Genome-scale model of C. autoethanogenum reveals optimal bioprocess conditions for high-value chemical production from carbon monoxide.

Engineering of vitamin prototrophy in Clostridium ljungdahlii and Clostridium autoethanogenum (2019)
Journal Article
Annan, F. J., Al-Sinawi, B., Humphreys, C. M., Norman, R., Winzer, Köpke, M., …Henstra, A. M. (2019). Engineering of vitamin prototrophy in Clostridium ljungdahlii and Clostridium autoethanogenum. Applied Microbiology and Biotechnology, 103(11), 4633–4648. https://doi.org/10.1007/s00253-019-09763-6

Clostridium autoethanogenum and Clostridium ljungdahlii are physiologically and genetically very similar strict anaerobic acetogens capable of growth on carbon monoxide as sole carbon source. While exact nutritional requirements have not been reporte... Read More about Engineering of vitamin prototrophy in Clostridium ljungdahlii and Clostridium autoethanogenum.