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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.

Base editing enables duplex point mutagenesis in Clostridium autoethanogenum at the price of numerous off-target mutations (2023)
Journal Article
Seys, F. M., Humphreys, C. M., Tomi-Andrino, C., Li, Q., Millat, T., Yang, S., & Minton, N. P. (2023). Base editing enables duplex point mutagenesis in Clostridium autoethanogenum at the price of numerous off-target mutations. Frontiers in Bioengineering and Biotechnology, 11, Article 1211197. https://doi.org/10.3389/fbioe.2023.1211197

Base editors are recent multiplex gene editing tools derived from the Cas9 nuclease of Streptomyces pyogenes. They can target and modify a single nucleotide in the genome without inducing double-strand breaks (DSB) of the DNA helix. As such, they hol... Read More about Base editing enables duplex point mutagenesis in Clostridium autoethanogenum at the price of numerous off-target mutations.

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.

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.

Establishing Mixotrophic Growth of Cupriavidus necator H16 on CO2 and Volatile Fatty Acids (2022)
Journal Article
Jawed, K., Irorere, V. U., Bommareddy, R. R., Minton, N. P., & Kovács, K. (2022). Establishing Mixotrophic Growth of Cupriavidus necator H16 on CO2 and Volatile Fatty Acids. Fermentation, 8(3), Article 125. https://doi.org/10.3390/fermentation8030125

The facultative chemolithoautotroph Cupriavidus necator H16 is able to grow aerobically either with organic substrates or H2 and CO2 s and it can accumulate large amounts of (up to 90%) poly (3‐hydroxybutyrate), a polyhydroxyalkanoate (PHA) biopolyme... Read More about Establishing Mixotrophic Growth of Cupriavidus necator H16 on CO2 and Volatile Fatty Acids.

Biosensor-informed engineering of Cupriavidus necator H16 for autotrophic D-mannitol production (2022)
Journal Article
Hanko, E. K., Sherlock, G., Minton, N. P., & Malys, N. (2022). Biosensor-informed engineering of Cupriavidus necator H16 for autotrophic D-mannitol production. Metabolic Engineering, 72, 24-34. https://doi.org/10.1016/j.ymben.2022.02.003

Cupriavidus necator H16 is one of the most researched carbon dioxide (CO2)-fixing bacteria. It can store carbon in form of the polymer polyhydroxybutyrate and generate energy by aerobic hydrogen oxidation under lithoautotrophic conditions, making C.... Read More about Biosensor-informed engineering of Cupriavidus necator H16 for autotrophic D-mannitol production.

A Novel Bacteriophage with Broad Host Range against Clostridioides difficile Ribotype 078 Supports SlpA as the Likely Phage Receptor (2022)
Journal Article
Whittle, M. J., Bilverstone, T. W., Van Esveld, R. J., Lücke, A. C., Lister, M. M., Kuehne, S. A., & Minton, N. P. (2022). A Novel Bacteriophage with Broad Host Range against Clostridioides difficile Ribotype 078 Supports SlpA as the Likely Phage Receptor. Microbiology Spectrum, 10(1), Article e02295-21. https://doi.org/10.1128/spectrum.02295-21

Bacteriophages represent a promising option for the treatment of Clostridioides difficile (formerly Clostridium difficile) infection (CDI), which at present relies on conventional antibiotic therapy. The specificity of bacteriophages should prevent d... Read More about A Novel Bacteriophage with Broad Host Range against Clostridioides difficile Ribotype 078 Supports SlpA as the Likely Phage Receptor.

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.

Biosynthesis of Poly(3HB- co-3HP) with Variable Monomer Composition in Recombinant Cupriavidus necator H16 (2021)
Journal Article
McGregor, C., Minton, N. P., & Kovács, K. (2021). Biosynthesis of Poly(3HB- co-3HP) with Variable Monomer Composition in Recombinant Cupriavidus necator H16. ACS Synthetic Biology, 10(12), 3343-3352. https://doi.org/10.1021/acssynbio.1c00283

Polyhydroxyalkanoates are attractive alternatives to traditional plastics. However, although polyhydroxybutyrate (PHB) is produced in large quantities by Cupriavidus necator H16, its properties are far from ideal for the manufacture of plastic produc... Read More about Biosynthesis of Poly(3HB- co-3HP) with Variable Monomer Composition in Recombinant Cupriavidus necator H16.

The pMTL70000 modular, plasmid vector series for strain engineering in Cupriavidus necator H16 (2021)
Journal Article
Ehsaan, M., Baker, J., Kovács, K., Malys, N., & Minton, N. P. (2021). The pMTL70000 modular, plasmid vector series for strain engineering in Cupriavidus necator H16. Journal of Microbiological Methods, 189, Article 106323. https://doi.org/10.1016/j.mimet.2021.106323

Cupriavidus necator H16 can convert CO2 into industrial chemicals and fuels. To facilitate its engineering, we designed, built and tested the pMTL70000 modular plasmids comprising standardised Cupriavidus and E. coli replicons, selectable markers and... Read More about The pMTL70000 modular, plasmid vector series for strain engineering in Cupriavidus necator H16.

What's a SNP between friends: The lineage of Clostridioides difficile R20291 can effect research outcomes (2021)
Journal Article
Monteford, J., Bilverstone, T. W., Ingle, P., Philip, S., Kuehne, S. A., & Minton, N. P. (2021). What's a SNP between friends: The lineage of Clostridioides difficile R20291 can effect research outcomes. Anaerobe, 71, Article 102422. https://doi.org/10.1016/j.anaerobe.2021.102422

Clostridioides difficile R20291 is the most studied PCR-Ribotype 027 isolate. The two predominant lineages of this hypervirulent strain, however, exhibit substantive phenotypic differences and possess genomes that differ by a small number of nucleoti... Read More about What's a SNP between friends: The lineage of Clostridioides difficile R20291 can effect research outcomes.

Engineering improved ethylene production: Leveraging systems Biology and adaptive laboratory evolution (2021)
Journal Article
Vaud, S., Pearcy, N., Hanževački, M., Van Hagen, A. M., Abdelrazig, S., Safo, L., …Bryan, S. J. (2021). Engineering improved ethylene production: Leveraging systems Biology and adaptive laboratory evolution. Metabolic Engineering, 67, 308-320. https://doi.org/10.1016/j.ymben.2021.07.001

Ethylene is a small hydrocarbon gas widely used in the chemical industry. Annual worldwide production currently exceeds 150 million tons, producing considerable amounts of CO2 contributing to climate change. The need for a sustainable alternative is... Read More about Engineering improved ethylene production: Leveraging systems Biology and adaptive laboratory evolution.

Development of a Suite of Tools for Genome Editing in Parageobacillus thermoglucosidasius and Their Use to Identify the Potential of a Native Plasmid in the Generation of Stable Engineered Strains (2021)
Journal Article
Lau, M. S. H., Sheng, L., Zhang, Y., & Minton, N. P. (2021). Development of a Suite of Tools for Genome Editing in Parageobacillus thermoglucosidasius and Their Use to Identify the Potential of a Native Plasmid in the Generation of Stable Engineered Strains. ACS Synthetic Biology, 10(7), 1739-1749. https://doi.org/10.1021/acssynbio.1c00138

The relentless rise in the levels of atmospheric greenhouse gases caused by the exploitation of fossil fuel necessitates the development of more environmentally friendly routes to the manufacture of chemicals and fuels. The exploitation of a fermenta... Read More about Development of a Suite of Tools for Genome Editing in Parageobacillus thermoglucosidasius and Their Use to Identify the Potential of a Native Plasmid in the Generation of Stable Engineered Strains.

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.

Colonisation factor cd0873, an attractive oral vaccine candidate against clostridioides difficile (2021)
Journal Article
Karyal, C., Hughes, J., Kelly, M. L., Cockayne, A., Luckett, J. C., Kaye, P. V., …Griffin, R. (2021). Colonisation factor cd0873, an attractive oral vaccine candidate against clostridioides difficile. Microorganisms, 9(2), Article 306. https://doi.org/10.3390/microorganisms9020306

Clostridioides difficile is the main cause of health-care-associated infectious diarrhoea. Toxins, TcdA and TcdB, secreted by this bacterium damage colonic epithelial cells and in severe cases this culminates in pseudomembranous colitis, toxic megaco... Read More about Colonisation factor cd0873, an attractive oral vaccine candidate against clostridioides difficile.

Variability in Arsenic Methylation Efficiency across Aerobic and Anaerobic Microorganisms (2020)
Journal Article
Viacava, K., Meibom, K. L., Ortega, D., Dyer, S., Gelb, A., Falquet, L., …Bernier-Latmani, R. (2020). Variability in Arsenic Methylation Efficiency across Aerobic and Anaerobic Microorganisms. Environmental Science and Technology, 54(22), 14343-14351. https://doi.org/10.1021/acs.est.0c03908

© 2020 American Chemical Society. Microbially-mediated methylation of arsenic (As) plays an important role in the As biogeochemical cycle, particularly in rice paddy soils where methylated As, generated microbially, is translocated into rice grains.... Read More about Variability in Arsenic Methylation Efficiency across Aerobic and Anaerobic Microorganisms.

Synthetic biology on acetogenic bacteria for highly efficient conversion of c1 gases to biochemicals (2020)
Journal Article
Jin, S., Bae, J., Song, Y., Pearcy, N., Shin, J., Kang, S., …Cho, B. K. (2020). Synthetic biology on acetogenic bacteria for highly efficient conversion of c1 gases to biochemicals. International Journal of Molecular Sciences, 21(20), Article 7639. https://doi.org/10.3390/ijms21207639

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. Synthesis gas, which is mainly produced from fossil fuels or biomass gasification, consists of C1 gases such as carbon monoxide, carbon dioxide, and methane as well as hydrogen. Acetogenic bac... Read More about Synthetic biology on acetogenic bacteria for highly efficient conversion of c1 gases to biochemicals.

The glucosyltransferase activity of C. difficile toxin b is required for disease pathogenesis (2020)
Journal Article
Bilverstone, T. W., Garland, M., Cave, R. J., Kelly, M. L., Tholen, M., Bouley, D. M., …Melnyk, R. A. (2020). The glucosyltransferase activity of C. difficile toxin b is required for disease pathogenesis. PLoS Pathogens, 16(9), Article e1008852. https://doi.org/10.1371/journal.ppat.1008852

© 2020 Bilverstone 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 ar... Read More about The glucosyltransferase activity of C. difficile toxin b is required for disease pathogenesis.

Using singular perturbation theory to determine kinetic parameters in a non-standard coupled enzyme assay (2020)
Journal Article
Dalwadi, M. P., Orol, D., Walter, F., Minton, N. P., King, J. R., & Kovács, K. (2020). Using singular perturbation theory to determine kinetic parameters in a non-standard coupled enzyme assay. Journal of Mathematical Biology, 81(2), 649-690. https://doi.org/10.1007/s00285-020-01524-8

We investigate how to characterize the kinetic parameters of an aminotransaminase using a non-standard coupled (or auxiliary) enzyme assay, where the peculiarity arises for two reasons. First, one of the products of the auxiliary enzyme is a substrat... Read More about Using singular perturbation theory to determine kinetic parameters in a non-standard coupled enzyme assay.

A Sustainable Chemicals Manufacturing Paradigm Using CO2 and Renewable H2 (2020)
Journal Article
Bommareddy, R. R., Wang, Y., Pearcy, N., Hayes, M., Lester, E., Minton, N. P., & Conradie, A. V. (2020). A Sustainable Chemicals Manufacturing Paradigm Using CO2 and Renewable H2. iScience, 23(6), https://doi.org/10.1016/j.isci.2020.101218

© 2020 The Author(s) The chemical industry must decarbonize to align with UN Sustainable Development Goals. A shift toward circular economies makes CO2 an attractive feedstock for producing chemicals, provided renewable H2 is available through techno... Read More about A Sustainable Chemicals Manufacturing Paradigm Using CO2 and Renewable H2.

A gold standard, CRISPR/Cas9-based complementation strategy reliant on 24 nucleotide bookmark sequences (2020)
Journal Article
Seys, F. M., Rowe, P., Bolt, E. L., Humphreys, C. M., & Minton, N. P. (2020). A gold standard, CRISPR/Cas9-based complementation strategy reliant on 24 nucleotide bookmark sequences. Genes, 11(4), Article 458. https://doi.org/10.3390/genes11040458

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. Phenotypic complementation of gene knockouts is an essential step in establishing function. Here, we describe a simple strategy for ‘gold standard’ complementation in which the mutant allele i... Read More about A gold standard, CRISPR/Cas9-based complementation strategy reliant on 24 nucleotide bookmark sequences.

A genome-wide approach for identification and characterisation of metabolite-inducible systems (2020)
Journal Article
Hanko, E. K. R., Paiva, A. C., Jonczyk, M., Abbott, M., Minton, N. P., & Malys, N. (2020). A genome-wide approach for identification and characterisation of metabolite-inducible systems. Nature Communications, 11(1), https://doi.org/10.1038/s41467-020-14941-6

© 2020, The Author(s). Inducible gene expression systems are vital tools for the advancement of synthetic biology. Their application as genetically encoded biosensors has the potential to contribute to diagnostics and to revolutionise the field of mi... Read More about A genome-wide approach for identification and characterisation of metabolite-inducible systems.

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.

Engineering Geobacillus thermoglucosidasius for direct utilisation of holocellulose from wheat straw (2019)
Journal Article
Bashir, Z., Sheng, L., Anil, A., Lali, A., Minton, N. P., & Zhang, Y. (2019). Engineering Geobacillus thermoglucosidasius for direct utilisation of holocellulose from wheat straw. Biotechnology for Biofuels, 12(1), Article 199. https://doi.org/10.1186/s13068-019-1540-6

Background: A consolidated bioprocessing (CBP), where lignocellulose is converted into the desired product(s) in a single fermentative step without the addition of expensive degradative enzymes, represents the ideal solution of renewable routes to ch... Read More about Engineering Geobacillus thermoglucosidasius for direct utilisation of holocellulose from wheat straw.

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.

Phosphorylation and functionality of CdtR in Clostridium difficile (2019)
Journal Article
Bilverstone, T., Minton, N., & Kuehne, S. (2019). Phosphorylation and functionality of CdtR in Clostridium difficile. Anaerobe, 58, 103-109. https://doi.org/10.1016/j.anaerobe.2019.102074

The production of TcdA, TcdB and CDT in Clostridium difficile PCR ribotype 027, is regulated by the two-component system response regulator CdtR. Despite this, little is known about the signal transduction pathway leading to the activation of CdtR. I... Read More about Phosphorylation and functionality of CdtR in Clostridium difficile.

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.

RiboCas: A Universal CRISPR-Based Editing Tool for Clostridium (2019)
Journal Article
Cañadas, I. C., Groothuis, D., Zygouropoulou, M., Rodrigues, R., & Minton, N. P. (2019). RiboCas: A Universal CRISPR-Based Editing Tool for Clostridium. ACS Synthetic Biology, 8(6), 1379-1390. https://doi.org/10.1021/acssynbio.9b00075

Members of the genus Clostridium represent a diverse assemblage of species exhibiting both medical and industrial importance. Deriving both a greater understanding of their biology, while at the same time enhancing their exploitable properties, requi... Read More about RiboCas: A Universal CRISPR-Based Editing Tool for Clostridium.

Generation of a fully erythromycin-sensitive strain of Clostridioides difficile using a novel CRISPR-Cas9 genome editing system (2019)
Journal Article
Ingle, P., Groothuis, D., Rowe, P., Huang, H., Cockayne, A., Kuehne, S. A., …Minton, N. P. (2019). Generation of a fully erythromycin-sensitive strain of Clostridioides difficile using a novel CRISPR-Cas9 genome editing system. Scientific Reports, 9, Article 8123. https://doi.org/10.1038/s41598-019-44458-y

© 2019, The Author(s). Understanding the molecular pathogenesis of Clostridioides difficile has relied on the use of ermB-based mutagens in erythromycin-sensitive strains. However, the repeated subcultures required to isolate sensitive variants can l... Read More about Generation of a fully erythromycin-sensitive strain of Clostridioides difficile using a novel CRISPR-Cas9 genome editing system.

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.

Homologous overexpression of hydrogenase and glycerol dehydrogenase in Clostridium pasteurianum to enhance hydrogen production from crude glycerol (2019)
Journal Article
Sarma, S., Ortega, D., Minton, N. P., Dubey, V. K., & Moholkar, V. S. (2019). Homologous overexpression of hydrogenase and glycerol dehydrogenase in Clostridium pasteurianum to enhance hydrogen production from crude glycerol. Bioresource Technology, 284, 168-177. https://doi.org/10.1016/j.biortech.2019.03.074

This study reports engineering of a hypertransformable variant of C. pasteurianum for bioconversion of glycerol into hydrogen (H2). A functional glycerol-triggered hydrogen pathway was engineered based on two approaches: (1) increasing product yield... Read More about Homologous overexpression of hydrogenase and glycerol dehydrogenase in Clostridium pasteurianum to enhance hydrogen production from crude glycerol.

Gsmodutils: a python based framework for test-driven genome scale metabolic model development (2019)
Journal Article
Gilbert, J., Pearcy, N., Norman, R., Millat, T., Winzer, K., King, J., …Twycross, J. (2019). Gsmodutils: a python based framework for test-driven genome scale metabolic model development. Bioinformatics, 35(18), 3397-3403. https://doi.org/10.1093/bioinformatics/btz088

© 2019 The Author(s) 2019. Published by Oxford University Press. Motivation: Genome scale metabolic models (GSMMs) are increasingly important for systems biology and metabolic engineering research as they are capable of simulating complex steady-stat... Read More about Gsmodutils: a python based framework for test-driven genome scale metabolic model development.

CRISPR-Cas9D10A nickase-assisted base editing in the solvent producer Clostridium beijerinckii (2019)
Journal Article
Li, Q., Seys, F. M., Minton, N. P., Yang, J., Jiang, Y., Jiang, W., & Yang, S. (2019). CRISPR-Cas9D10A nickase-assisted base editing in the solvent producer Clostridium beijerinckii. Biotechnology and Bioengineering, 116(6), 1475-1483. https://doi.org/10.1002/bit.26949

Clostridium beijerinckii is a potentially important industrial microorganism as it can synthesize valuable chemicals and fuels from various carbon sources. The establishment of convenient to use, effective gene tools with which the organism can be ra... Read More about CRISPR-Cas9D10A nickase-assisted base editing in the solvent producer Clostridium beijerinckii.

The Rnf complex is an energy-coupled transhydrogenase essential to reversibly link cellular NADH and ferredoxin pools in the acetogen acetobacterium woodii (2018)
Journal Article
Westphal, L., Wiechmann, A., Baker, J., Minton, N. P., & Müller, V. (2018). The Rnf complex is an energy-coupled transhydrogenase essential to reversibly link cellular NADH and ferredoxin pools in the acetogen acetobacterium woodii. Journal of Bacteriology, 200(21), https://doi.org/10.1128/JB.00357-18

© 2018 American Society for Microbiology. The Rnf complex is a respiratory enzyme that catalyzes the oxidation of reduced ferredoxin to the reduction of NAD + , and the negative free energy change of this reaction is used to generate a transmembrane... Read More about The Rnf complex is an energy-coupled transhydrogenase essential to reversibly link cellular NADH and ferredoxin pools in the acetogen acetobacterium woodii.

Gsmodutils: A python based framework for test-driven genome scale metabolic model development (2018)
Other
Gilbert, J. P., Pearcy, N., Norman, R., Millat, T., Winzer, K., King, J., …Twycross, J. (2018). Gsmodutils: A python based framework for test-driven genome scale metabolic model development

Motivation Genome scale metabolic models (GSMMs) are increasingly important for systems biology and metabolic engineering research as they are capable of simulating complex steady-state behaviour. Constraints based models of this form can include tho... Read More about Gsmodutils: A python based framework for test-driven genome scale metabolic model development.

Functional genetic elements for controlling gene expression in Cupriavidus necator H16 (2018)
Journal Article
Alagesan, S., Hanko, E. K., Malys, N., Ehsaan, M., Winzer, K., & Minton, N. P. (2018). Functional genetic elements for controlling gene expression in Cupriavidus necator H16. Applied and Environmental Microbiology, https://doi.org/10.1128/AEM.00878-18

A robust and predictable control of gene expression plays an important role in synthetic biology and biotechnology applications. Development and quantitative evaluation of functional genetic elements such as constitutive and inducible promoters, as w... Read More about Functional genetic elements for controlling gene expression in Cupriavidus necator H16.

Effect of antibiotic treatment on the formation of non-spore Clostridium difficile persister-like cells (2018)
Journal Article
Álvarez, R., Inostroza, O., Garavaglia, M., Minton, N. P., Paredes-Sabja, D., & Gil, F. (2018). Effect of antibiotic treatment on the formation of non-spore Clostridium difficile persister-like cells. Journal of Antimicrobial Chemotherapy, 73(9), 2396-2399. https://doi.org/10.1093/jac/dky186

Background: The spore is the virulence factor identified to be involved in the recurrence of the disease caused by Clostridium difficile. Objectives: To demonstrate that lethal antibiotic concentrations induce the appearance of C. difficile persister... Read More about Effect of antibiotic treatment on the formation of non-spore Clostridium difficile persister-like cells.

The butanol producing microbe Clostridium beijerinckii NCIMB 14988 manipulated using forward and reverse genetic tools (2018)
Journal Article
Little, G. T., Willson, B. J., Heap, J. T., Winzer, K., & Minton, N. P. (2018). The butanol producing microbe Clostridium beijerinckii NCIMB 14988 manipulated using forward and reverse genetic tools. Biotechnology Journal, https://doi.org/10.1002/biot.201700711

The solventogenic anaerobe Clostridium beijerinckii has potential for use in the sustainable bioconversion of plant-derived carbohydrates into solvents, such as butanol or acetone. However, relatively few strains have been extensively characterised e... Read More about The butanol producing microbe Clostridium beijerinckii NCIMB 14988 manipulated using forward and reverse genetic tools.

A transcription factor-based biosensor for detection of itaconic acid (2018)
Journal Article
Hanko, E. K., Minton, N. P., & Malys, N. (2018). A transcription factor-based biosensor for detection of itaconic acid. ACS Synthetic Biology, 7(5), 1436–1446. https://doi.org/10.1021/acssynbio.8b00057

Itaconic acid is an important platform chemical that can easily be incorporated into polymers and has the potential to replace petrochemical-based acrylic or methacrylic acid. A number of microorganisms have been developed for the biosynthesis of ita... Read More about A transcription factor-based biosensor for detection of itaconic acid.

Quantitative isotope-dilution high-resolution-mass-apectrometry analysis of multiple intracellular metabolites in Clostridium autoethanogenum with uniformly 13C-labeled standards derived from Spirulina (2018)
Journal Article
Schatschneider, S., Abdelrazig, S. M., Safo, L., Henstra, A. M., Millat, T., Kim, D., …Barrett, D. A. (2018). Quantitative isotope-dilution high-resolution-mass-apectrometry analysis of multiple intracellular metabolites in Clostridium autoethanogenum with uniformly 13C-labeled standards derived from Spirulina. Analytical Chemistry, 90(7), https://doi.org/10.1021/acs.analchem.7b04758

We have investigated the applicability of commercially available lyophilized spirulina (Arthrospira platensis), a microorganism uniformly labeled with 13C, as a readily accessible source of multiple 13C-labeled metabolites suitable as internal standa... Read More about Quantitative isotope-dilution high-resolution-mass-apectrometry analysis of multiple intracellular metabolites in Clostridium autoethanogenum with uniformly 13C-labeled standards derived from Spirulina.

Advances in metabolic engineering in the microbial production of fuels and chemicals from C1 gas (2018)
Journal Article
Humphreys, C. M., & Minton, N. P. (in press). Advances in metabolic engineering in the microbial production of fuels and chemicals from C1 gas. Current Opinion in Biotechnology, 50, https://doi.org/10.1016/j.copbio.2017.12.023

The future sustainable production of chemicals and fuels from non-petrochemical sources, while at the same time reducing greenhouse gas (GHG) emissions, represent two of society's greatest challenges. Microbial chassis able to grow on waste carbon mo... Read More about Advances in metabolic engineering in the microbial production of fuels and chemicals from C1 gas.

Upscaling Diffusion Through First-Order Volumetric Sinks: A Homogenization of Bacterial Nutrient Uptake (2018)
Journal Article
Dalwadi, M. P., Wang, Y., King, J. R., & Minton, N. P. (2018). Upscaling Diffusion Through First-Order Volumetric Sinks: A Homogenization of Bacterial Nutrient Uptake. SIAM Journal on Applied Mathematics, 78(3), 1300-1329. https://doi.org/10.1137/17m1138625

In mathematical models that include nutrient delivery to bacteria, it is prohibitively expensive to include a pointwise nutrient uptake within small bacterial regions over bioreactor length-scales, and so such models often impose an effective uptake... Read More about Upscaling Diffusion Through First-Order Volumetric Sinks: A Homogenization of Bacterial Nutrient Uptake.

Characterization of the impact of rpoB mutations on the in vitro and in vivo competitive fitness of Clostridium difficile and susceptibility to fidaxomicin (2017)
Journal Article
Kuehne, S. A., Dempster, A. W., Collery, M. M., Joshi, N., Jowett, J., Kelly, M. L., …Minton, N. P. (2018). Characterization of the impact of rpoB mutations on the in vitro and in vivo competitive fitness of Clostridium difficile and susceptibility to fidaxomicin. Journal of Antimicrobial Chemotherapy, 73(4), 973-980. https://doi.org/10.1093/jac/dkx486

Objectives To establish the role of specific, non-synonymous SNPs in the RNA polymerase ? subunit (rpoB) gene in reducing the susceptibility of Clostridium difficile to fidaxomicin and to explore the potential in vivo significance of rpoB mutant str... Read More about Characterization of the impact of rpoB mutations on the in vitro and in vivo competitive fitness of Clostridium difficile and susceptibility to fidaxomicin.

13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16 (2017)
Journal Article
Alagesan, S., Minton, N. P., & Malys, N. (2018). 13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16. Metabolomics, 14(9), https://doi.org/10.1007/s11306-017-1302-z

Introduction. Cupriavidus necator H16 is a gram-negative bacterium, capable of lithoautotrophic growth by utilizing hydrogen as an energy source and fixing carbon dioxide (CO2) through Calvin-Benson-Bassham (CBB) cycle. The potential to utilize synth... Read More about 13C-assisted metabolic flux analysis to investigate heterotrophic and mixotrophic metabolism in Cupriavidus necator H16.

Applying asymptotic methods to synthetic biology: modelling the reaction kinetics of the mevalonate pathway (2017)
Journal Article
Dalwadi, M. P., Garavaglia, M., Webb, J. P., King, J. R., & Minton, N. P. (2018). Applying asymptotic methods to synthetic biology: modelling the reaction kinetics of the mevalonate pathway. Journal of Theoretical Biology, 439, https://doi.org/10.1016/j.jtbi.2017.11.022

The mevalonate pathway is normally found in eukaryotes, and allows for the production of isoprenoids, a useful class of organic compounds. This pathway has been successfully introduced to Escherichia coli, enabling a biosynthetic production route for... Read More about Applying asymptotic methods to synthetic biology: modelling the reaction kinetics of the mevalonate pathway.

Multi-timescale analysis of a metabolic network in synthetic biology: a kinetic model for 3-hydroxypropionic acid production via beta-alanine (2017)
Journal Article
Dalwadi, M. P., King, J. R., & Minton, N. P. (2018). Multi-timescale analysis of a metabolic network in synthetic biology: a kinetic model for 3-hydroxypropionic acid production via beta-alanine. Journal of Mathematical Biology, 77(1), 165-199. https://doi.org/10.1007/s00285-017-1189-3

A biosustainable production route for 3-hydroxypropionic acid (3HP), an important platform chemical, would allow 3HP to be produced without using fossil fuels. We are interested in investigating a potential biochemical route to 3HP from pyruvate thro... Read More about Multi-timescale analysis of a metabolic network in synthetic biology: a kinetic model for 3-hydroxypropionic acid production via beta-alanine.

Recombinant expression and characterisation of the oxygen-sensitive 2-enoate reductase from Clostridium sporogenes (2017)
Journal Article
Mordaka, P. M., Hall, S. J., Minton, N. P., & Stephens, G. (in press). Recombinant expression and characterisation of the oxygen-sensitive 2-enoate reductase from Clostridium sporogenes. Microbiology, https://doi.org/10.1099/mic.0.000568

‘Ene’-reductases have attracted significant attention for the preparation of chemical intermediates and biologically active products. To date, research has been focussed primarily on Old Yellow Enzyme-like proteins, due to their ease of handling, whe... Read More about Recombinant expression and characterisation of the oxygen-sensitive 2-enoate reductase from Clostridium sporogenes.

Improving gene transfer in Clostridium pasteurianum through the isolation of rare hypertransformable variants (2017)
Journal Article
Grosse-Honebrink, A., Schwarz, K., Wang, H., Minton, N. P., & Zhang, Y. (2017). Improving gene transfer in Clostridium pasteurianum through the isolation of rare hypertransformable variants. Anaerobe, 48, https://doi.org/10.1016/j.anaerobe.2017.09.001

Effective microbial metabolic engineering is reliant on efficient gene transfer. Here we present a simple screening strategy that may be deployed to isolate rare, hypertransformable variants. The procedure was used to increase the frequency of transf... Read More about Improving gene transfer in Clostridium pasteurianum through the isolation of rare hypertransformable variants.

Exposing hidden putative lipoproteins in Clostridium difficile (2017)
Presentation / Conference
Griffin, R., & Minton, N. P. (2017, August). Exposing hidden putative lipoproteins in Clostridium difficile. Poster presented at ClostPath 10th International Conference on the Molecular Biology and Pathogenesis of the Clostrida, Ann Arbor, Michigan, USA

Characterisation of a 3-hydroxypropionic acid-inducible system from Pseudomonas putida for orthogonal gene expression control in Escherichia coli and Cupriavidus necator (2017)
Journal Article
Hanko, E. K., Minton, N. P., & Malys, N. (in press). Characterisation of a 3-hydroxypropionic acid-inducible system from Pseudomonas putida for orthogonal gene expression control in Escherichia coli and Cupriavidus necator. Scientific Reports, 7, Article 1724. https://doi.org/10.1038/s41598-017-01850-w

3-hydroxypropionic acid (3-HP) is an important platform chemical used as a precursor for production of added-value compounds such as acrylic acid. Metabolically engineered yeast, Escherichia coli, cyanobacteria and other microorganisms have been deve... Read More about Characterisation of a 3-hydroxypropionic acid-inducible system from Pseudomonas putida for orthogonal gene expression control in Escherichia coli and Cupriavidus necator.

Microbial solvent formation revisited by comparative genome analysis (2017)
Journal Article
Poehlein, A., Solano, J. D. M., Flitsch, S. K., Krabben, P., Winzer, K., Reid, S. J., …Dürre, P. (2017). Microbial solvent formation revisited by comparative genome analysis. Biotechnology for Biofuels, 10, Article 58. https://doi.org/10.1186/s13068-017-0742-z

Background: Microbial formation of acetone, isopropanol, and butanol is largely restricted to bacteria belonging to the genus Clostridium. This ability has been industrially exploited over the last 100 years. The solvents are important feedstocks for... Read More about Microbial solvent formation revisited by comparative genome analysis.

Towards improved butanol production through targeted genetic modification of Clostridium pasteurianum (2017)
Journal Article
Schwarz, K., Grosse-Honebrink, A., Derecka, K., Rotta, C., Zhang, Y., & Minton, N. P. (2017). Towards improved butanol production through targeted genetic modification of Clostridium pasteurianum. Metabolic Engineering, 40, 124-137. https://doi.org/10.1016/j.ymben.2017.01.009

Declining fossil fuel reserves, coupled with environmental concerns over their continued extraction and exploitation have led to strenuous efforts to identify renewable routes to energy and fuels. One attractive option is to convert glycerol, a by-pr... Read More about Towards improved butanol production through targeted genetic modification of Clostridium pasteurianum.

Metabolic engineering of Clostridium autoethanogenum for selective alcohol production (2017)
Journal Article
Liew, F., Henstra, A. M., Kӧpke, M., Winzer, K., Simpson, S. D., & Minton, N. P. (2017). Metabolic engineering of Clostridium autoethanogenum for selective alcohol production. Metabolic Engineering, 40, 104-114. https://doi.org/10.1016/j.ymben.2017.01.007

Gas fermentation using acetogenic bacteria such as Clostridium autoethanogenum offers an attractive route for production of fuel ethanol from industrial waste gases. Acetate reduction to acetaldehyde and further to ethanol via an aldehyde: ferredoxin... Read More about Metabolic engineering of Clostridium autoethanogenum for selective alcohol production.

Development of Clostridium difficile R20291?PaLoc model strains and in vitro methodologies reveals CdtR is required for the production of CDT to cytotoxic levels (2017)
Journal Article
Bilverstone, T., Kinsmore, N., Minton, N. P., & Kuehne, S. A. (in press). Development of Clostridium difficile R20291?PaLoc model strains and in vitro methodologies reveals CdtR is required for the production of CDT to cytotoxic levels. Anaerobe, 44, https://doi.org/10.1016/j.anaerobe.2017.01.009

Assessing the regulation of Clostridium difficile transferase (CDT), is complicated by the presence of a Pathogenicity locus (PaLoc) which encodes Toxins A and B. Here we developed R20291?PaLoc model strains and cell-based assays to quantify CDT-medi... Read More about Development of Clostridium difficile R20291?PaLoc model strains and in vitro methodologies reveals CdtR is required for the production of CDT to cytotoxic levels.

What's a SNP between friends: The influence of single nucleotide polymorphisms on virulence and phenotypes of Clostridium difficile strain 630 and derivatives (2016)
Journal Article
Collery, M. M., Kuehne, S. A., McBride, S. M., Kelly, M. L., Monot, M., Cockayne, A., …Minton, N. P. (2017). What's a SNP between friends: The influence of single nucleotide polymorphisms on virulence and phenotypes of Clostridium difficile strain 630 and derivatives. Virulence, 8(6), 767-781. https://doi.org/10.1080/21505594.2016.1237333

Clostridium difficile is a major cause of antibiotic induced diarrhoea worldwide, responsible for significant annual mortalities and represents a considerable economic burden on healthcare systems. The two main C. difficile virulence factors are toxi... Read More about What's a SNP between friends: The influence of single nucleotide polymorphisms on virulence and phenotypes of Clostridium difficile strain 630 and derivatives.

Coinfection and Emergence of Rifamycin Resistance during a Recurrent Clostridium difficile Infection (2016)
Journal Article
Stevenson, E. C., Major, G. A., Spiller, R. C., Kuehne, S. A., & Minton, N. P. (2016). Coinfection and Emergence of Rifamycin Resistance during a Recurrent Clostridium difficile Infection. Journal of Clinical Microbiology, 54(11), 2689-2694. https://doi.org/10.1128/jcm.01025-16

Clostridium difficile (Peptoclostridium difficile) is a common health care-associated infection with a disproportionately high incidence in elderly patients. Disease symptoms range from mild diarrhea to life-threatening pseudomembranous colitis. Arou... Read More about Coinfection and Emergence of Rifamycin Resistance during a Recurrent Clostridium difficile Infection.

A genetic assay for gene essentiality in Clostridium (2016)
Journal Article
Walker, D. J., Heap, J. T., Winzer, K., & Minton, N. P. (2016). A genetic assay for gene essentiality in Clostridium. Anaerobe, 42, 40-43. https://doi.org/10.1016/j.anaerobe.2016.07.007

Essential genes of pathogens are potential therapeutic targets, but are difficult to verify. Here, gene essentiality was determined by targeted knockout following engineered gene duplication. Null mutants of candidate essential genes of Clostridium d... Read More about A genetic assay for gene essentiality in Clostridium.

The binary toxin CDT enhances Clostridium difficile virulence by suppressing protective colonic eosinophilia (2016)
Journal Article
Cowardin, C. A., Buonomo, E. L., Saleh, M. M., Wilson, M. G., Burgess, S. L., Schwan, C., …Petri, W. A. (2016). The binary toxin CDT enhances Clostridium difficile virulence by suppressing protective colonic eosinophilia. Nature Microbiology, 1(8), Article 16108. https://doi.org/10.1038/nmicrobiol.2016.108

© 2016 Macmillan Publishers Limited. All rights reserved. Clostridium difficile is the most common hospital acquired pathogen in the USA, and infection is, in many cases, fatal. Toxins A and B are its major virulence factors, but expression of a thir... Read More about The binary toxin CDT enhances Clostridium difficile virulence by suppressing protective colonic eosinophilia.

SBRC-Nottingham: sustainable routes to platform chemicals from C1 wastes Gases (2016)
Journal Article
Burbidge, A., & Minton, N. P. (2016). SBRC-Nottingham: sustainable routes to platform chemicals from C1 wastes Gases. Biochemical Society Transactions, 44(3), 684-686. https://doi.org/10.1042/BST20160010

SBRC-Nottingham (www.sbrc-nottingham.ac.uk) was one of the first three UK university-based Synthetic Biology Research Centres (SBRCs) to be funded by the BBSRC and EPSRC as part of the recommendations made in the UKs Synthetic Biology Roadmap. It was... Read More about SBRC-Nottingham: sustainable routes to platform chemicals from C1 wastes Gases.

Insights into CO2 fixation pathway of Clostridium autoethanogenumby targeted mutagenesis (2016)
Journal Article
Liew, F., Henstra, A. M., Winzer, K., Kopke, M., Simpson, S. D., & Minton, N. P. (in press). Insights into CO2 fixation pathway of Clostridium autoethanogenumby targeted mutagenesis. mBio, 7(3), Article e00427-16. https://doi.org/10.1128/mBio.00427-16

The future sustainable production of chemicals and fuels from nonpetrochemical resources and reduction of greenhouse gas emissions are two of the greatest societal challenges. Gas fermentation, which utilizes the ability of acetogenic bacteria such a... Read More about Insights into CO2 fixation pathway of Clostridium autoethanogenumby targeted mutagenesis.

A roadmap for gene system development in Clostridium (2016)
Journal Article
Minton, N. P., Ehsaan, M., Humphreys, C. M., Little, G. T., Baker, J., Henstra, A. M., …Zhang, Y. (2016). A roadmap for gene system development in Clostridium. Anaerobe, 41, 104-112. https://doi.org/10.1016/j.anaerobe.2016.05.011

Clostridium species are both heroes and villains. Some cause serious human and animal diseases, those present in the microbiota contribute to health and wellbeing, while others represent useful industrial chassis for the production of chemicals and f... Read More about A roadmap for gene system development in Clostridium.

CRISPR-based genome editing and expression control systems in Clostridium acetobutylicum and Clostridium beijerinckii (2016)
Journal Article
Li, Q., Cheng, J., Minton, N. P., Zhang, Y., Wen, Z., Liu, J., …Yang, S. (2016). CRISPR-based genome editing and expression control systems in Clostridium acetobutylicum and Clostridium beijerinckii. Biotechnology Journal, 11(7), 961-972. https://doi.org/10.1002/biot.201600053

Solventogenic clostridia are important industrial microorganisms that produce various chemicals and fuels. Effective genetic tools would facilitate physiological studies aimed both at improving our understanding of metabolism and optimizing solvent p... Read More about CRISPR-based genome editing and expression control systems in Clostridium acetobutylicum and Clostridium beijerinckii.

Production of a functional cell wall-anchored minicellulosome by recombinant Clostridium acetobutylicum ATCC 824 (2016)
Journal Article
Willson, B. J., Kovács, K., Wilding-Steele, T., Markus, R., Winzer, K., & Minton, N. P. (2016). Production of a functional cell wall-anchored minicellulosome by recombinant Clostridium acetobutylicum ATCC 824. Biotechnology for Biofuels, 9(1), Article 109. https://doi.org/10.1186/s13068-016-0526-x

Background: The use of fossil fuels is no longer tenable. Not only are they a finite resource, their use is damaging the environment through pollution and global warming. Alternative, environmentally friendly, renewable sources of chemicals and fuels... Read More about Production of a functional cell wall-anchored minicellulosome by recombinant Clostridium acetobutylicum ATCC 824.

Development of an inducible transposon system for efficient random mutagenesis in Clostridium acetobutylicum (2016)
Journal Article
Zhang, Y., Xu, S., Chai, C., Yang, S., Jiang, W., Minton, N. P., & Gu, Y. (2016). Development of an inducible transposon system for efficient random mutagenesis in Clostridium acetobutylicum. FEMS Microbiology Letters, 363(8), Article fnw065. https://doi.org/10.1093/femsle/fnw065

Clostridium acetobutylicum is an industrially important Gram-positive organism which is capable of producing economically important chemicals in the ABE (Acetone, Butanol and Ethanol) fermentation process. Renewed interests in the ABE process necessi... Read More about Development of an inducible transposon system for efficient random mutagenesis in Clostridium acetobutylicum.

Improving the reproducibility of the NAP1/B1/027 epidemic strain R20291 in the hamster model of infection (2016)
Journal Article
Kelly, M. L., Ng, Y. K., Cartman, S. T., Collery, M. M., Cockayne, A., & Minton, N. P. (2016). Improving the reproducibility of the NAP1/B1/027 epidemic strain R20291 in the hamster model of infection. Anaerobe, 39, 51-53. https://doi.org/10.1016/j.anaerobe.2016.02.011

Comparative analysis of the Clostridium difficile BI/NAP1/027 strain R20291 and ClosTron-derived ermB mutants in the hamster infection model are compromised by the clindamycin susceptibility of the parent. Mutants can appear more virulent. We have re... Read More about Improving the reproducibility of the NAP1/B1/027 epidemic strain R20291 in the hamster model of infection.

Mutant generation by allelic exchange and genome resequencing of the biobutanol organism Clostridium acetobutylicum ATCC 824 (2016)
Journal Article
Ehsaan, M., Kuit, W., Zhang, Y., Cartman, S. T., Heap, J. T., Winzer, K., & Minton, N. P. (2016). Mutant generation by allelic exchange and genome resequencing of the biobutanol organism Clostridium acetobutylicum ATCC 824. Biotechnology for Biofuels, 9(1), Article 4. https://doi.org/10.1186/s13068-015-0410-0

Background Clostridium acetobutylicum represents a paradigm chassis for the industrial production of the biofuel biobutanol and a focus for metabolic engineering. We have previously developed procedures for the creation of in-frame, marker-less de... Read More about Mutant generation by allelic exchange and genome resequencing of the biobutanol organism Clostridium acetobutylicum ATCC 824.

Whole genome sequence and manual annotation of Clostridium autoethanogenum, an industrially relevant bacterium (2015)
Journal Article
Humphreys, C. M., McLean, S., Schatschneider, S., Millat, T., Henstra, A. M., Annan, F. J., …Minton, N. P. (2015). Whole genome sequence and manual annotation of Clostridium autoethanogenum, an industrially relevant bacterium. BMC Genomics, 16(1), Article 1085. https://doi.org/10.1186/s12864-015-2287-5

© 2015 Humphreys et al. Background: Clostridium autoethanogenum is an acetogenic bacterium capable of producing high value commodity chemicals and biofuels from the C1 gases present in synthesis gas. This common industrial waste gas can act as the so... Read More about Whole genome sequence and manual annotation of Clostridium autoethanogenum, an industrially relevant bacterium.

Clostridium difficile-mediated effects on human intestinal epithelia: Modelling host-pathogen interactions in a vertical diffusion chamber (2015)
Journal Article
Jafari, N. V., Kuehne, S. A., Minton, N. P., Allan, E., & Bajaj-Elliott, M. (2016). Clostridium difficile-mediated effects on human intestinal epithelia: Modelling host-pathogen interactions in a vertical diffusion chamber. Anaerobe, 37, 96-102. doi:10.1016/j.anaerobe.2015.12.007

Clostridium difficile infection is one of the leading causes of healthcare associated diarrhoea in the developed world. Although the contribution of C. difficile toxins to disease pathogenesis is now well understood, many facets of host-pathogen inte... Read More about Clostridium difficile-mediated effects on human intestinal epithelia: Modelling host-pathogen interactions in a vertical diffusion chamber.

A universal mariner transposon system for forward genetic studies in the genus Clostridium (2015)
Journal Article
Zhang, Y., Grosse-Honebrink, A., & Minton, N. P. (2015). A universal mariner transposon system for forward genetic studies in the genus Clostridium. PLoS ONE, 10(4), 1-21. https://doi.org/10.1371/journal.pone.0122411

DNA transposons represent an essential tool in the armoury of the molecular microbiologist. We previously developed a catP-based mini transposon system for Clostridium difficile in which the expression of the transposase gene was dependent on a sigma... Read More about A universal mariner transposon system for forward genetic studies in the genus Clostridium.

Coenzyme A-transferase-independent butyrate re-assimilation in Clostridium acetobutylicum - evidence from a mathematical model (2014)
Journal Article
Millat, T., Voigt, C., Janssen, H., Cooksley, C. M., Winzer, K., Minton, N. P., …Wolkenhauer, O. (2014). Coenzyme A-transferase-independent butyrate re-assimilation in Clostridium acetobutylicum - evidence from a mathematical model. Applied Microbiology and Biotechnology, 98(21), https://doi.org/10.1007/s00253-014-5987-x

The hetero-dimeric CoA-transferase CtfA/B is believed to be crucial for the metabolic transition from acidogenesis to solventogenesis in Clostridium acetobutylicum as part of the industrial-relevant acetone-butanol-ethanol (ABE) fermentation. Here, t... Read More about Coenzyme A-transferase-independent butyrate re-assimilation in Clostridium acetobutylicum - evidence from a mathematical model.

The role of flagella in Clostridium difficile pathogenesis:comparison between a non-epidemic and an epidemicstrain (2013)
Journal Article
Baban, S. T., Kuehne, S. A., Barketi-Klai, A., Cartman, S. T., Kelly, M. L., Hardie, K. R., …Minton, N. P. (2013). The role of flagella in Clostridium difficile pathogenesis:comparison between a non-epidemic and an epidemicstrain. PLoS ONE, 8(9), Article e73026. https://doi.org/10.1371/journal.pone.0073026

Clostridium difficile is a major cause of healthcare-associated infection and inflicts a considerable financial burden on healthcare systems worldwide. Disease symptoms range from self-limiting diarrhoea to fatal pseudomembranous colitis. Whilst C. d... Read More about The role of flagella in Clostridium difficile pathogenesis:comparison between a non-epidemic and an epidemicstrain.

Secretion and assembly of functional mini-cellulosomes from synthetic chromosomal operons in Clostridium acetobutylicum ATCC 824 (2013)
Journal Article
Kovacs, K., Willson, B. J., Schwarz, K., Heap, J. T., Jackson, A., Bolam, D. N., …Minton, N. P. (2013). Secretion and assembly of functional mini-cellulosomes from synthetic chromosomal operons in Clostridium acetobutylicum ATCC 824. Biotechnology for Biofuels, 6(117), https://doi.org/10.1186/1754-6834-6-117

BACKGROUND: Consolidated bioprocessing (CBP) is reliant on the simultaneous enzyme production, saccharification of biomass, and fermentation of released sugars into valuable products such as butanol. Clostridial species that produce butanol are, h... Read More about Secretion and assembly of functional mini-cellulosomes from synthetic chromosomal operons in Clostridium acetobutylicum ATCC 824.

Importance of Toxin A, Toxin B, and CDT in virulence of an epidemic Clostridium difficile strain (2013)
Journal Article
Kuehne, S. A., Collery, M. M., Kelly, M. L., Cartman, S. T., Cockayne, A., & Minton, N. P. (in press). Importance of Toxin A, Toxin B, and CDT in virulence of an epidemic Clostridium difficile strain. Journal of Infectious Diseases, 209(1), https://doi.org/10.1093/infdis/jit426

Clostridium difficile infection is the main cause of healthcare-acquired diarrhea in the developed world. In addition to the main virulence factors toxin A and B, epidemic, PCR Ribotype 027 strains, such as R20291, produce a third toxin, CDT. To deve... Read More about Importance of Toxin A, Toxin B, and CDT in virulence of an epidemic Clostridium difficile strain.

Clostridium difficile modulates host innate immunity via toxin-independent and dependent mechanism(s) (2013)
Journal Article
Jafari, N. V., Kuehne, S. A., Bryant, C. E., Elawad, M., Wren, B. W., Minton, N. P., …Bajaj-Elliott, M. (2013). Clostridium difficile modulates host innate immunity via toxin-independent and dependent mechanism(s). PLoS ONE, 8(7), Article e69846. https://doi.org/10.1371/journal.pone.0069846

Clostridium difficile infection (CDI) is the leading cause of hospital and community-acquired antibiotic-associated diarrhoea and currently represents a significant health burden. Although the role and contribution of C. difficile toxins to disease p... Read More about Clostridium difficile modulates host innate immunity via toxin-independent and dependent mechanism(s).

The putative influence of the agr operon upon survival mechanisms used by Clostridium acetobutylicum (2013)
Journal Article
Jabbari, S., Steiner, E., Heap, J. T., Winzer, K., Minton, N. P., & King, J. R. (2013). The putative influence of the agr operon upon survival mechanisms used by Clostridium acetobutylicum. Mathematical Biosciences, 243(2), https://doi.org/10.1016/j.mbs.2013.03.005

The bacterium Clostridium acetobutylicum produces acids as an energy-yielding process during exponential growth. An acidic environment, however, is toxic to the cells and two survival mechanisms are in place to prevent them from dying. Firstly, durin... Read More about The putative influence of the agr operon upon survival mechanisms used by Clostridium acetobutylicum.

Expanding the repertoire of gene tools for precise manipulation of the Clostridium difficile genome: allelic exchange using pyrE alleles (2013)
Journal Article
Ng, Y. K., Ehsaan, M., Philip, S., Collery, M. M., Janoir, C., Collignon, A., …Minton, N. P. (2013). Expanding the repertoire of gene tools for precise manipulation of the Clostridium difficile genome: allelic exchange using pyrE alleles. PLoS ONE, 8(2), Article 13. https://doi.org/10.1371/journal.pone.0056051

Sophisticated genetic tools to modify essential biological processes at the molecular level are pivotal in elucidating the molecular pathogenesis of Clostridium difficile, a major cause of healthcare associated disease. Here we have developed an effi... Read More about Expanding the repertoire of gene tools for precise manipulation of the Clostridium difficile genome: allelic exchange using pyrE alleles.

Multiple factors modulate biofilm formation by the anaerobic pathogen Clostridium difficile (2013)
Journal Article
Dapa, T., Leuzzi, R., Ng, Y. K., Baban, S. T., Adamo, R., Kuehne, S. A., …Unnikrishnan, M. (2013). Multiple factors modulate biofilm formation by the anaerobic pathogen Clostridium difficile. Journal of Bacteriology, 195(3), https://doi.org/10.1128/JB.01980-12

Bacteria within biofilms are protected from multiple stresses, including immune responses and antimicrobial agents. The biofilm-forming ability of bacterial pathogens has been associated with increased antibiotic resistance and chronic recurrent infe... Read More about Multiple factors modulate biofilm formation by the anaerobic pathogen Clostridium difficile.

Spores of Clostridium difficile clinical isolates display a diverse germination response to bile salts (2012)
Journal Article
Heeg, D., Burns, D. A., Cartman, S. T., & Minton, N. P. (2012). Spores of Clostridium difficile clinical isolates display a diverse germination response to bile salts. PLoS ONE, 7(2), Article e32381. https://doi.org/10.1371/journal.pone.0032381

Clostridium difficile spores play a pivotal role in the transmission of infectious diarrhoea, but in order to cause disease spores must complete germination and return to vegetative cell growth. While the mechanisms of spore germination are well unde... Read More about Spores of Clostridium difficile clinical isolates display a diverse germination response to bile salts.

Integration of DNA into bacterial chromosomes from plasmids without a counter-selection marker (2012)
Journal Article
Heap, J. T., Ehsaan, M., Cooksley, C. M., Ng, Y., Cartman, S. T., Winzer, K., & Minton, N. P. (2012). Integration of DNA into bacterial chromosomes from plasmids without a counter-selection marker. Nucleic Acids Research, 40(8), Article 10. https://doi.org/10.1093/nar/gkr1321

Most bacteria can only be transformed with circular plasmids, so robust DNA integration methods for these rely upon selection of single-crossover clones followed by counter-selection of double-crossover clones. To overcome the limited availability of... Read More about Integration of DNA into bacterial chromosomes from plasmids without a counter-selection marker.

Both, toxin A and toxin B, are important in Clostridium difficile infection (2011)
Journal Article
Kuehne, S. A., Cartman, S. T., & Minton, N. P. (2011). Both, toxin A and toxin B, are important in Clostridium difficile infection. Gut Microbes, 2(4), https://doi.org/10.4161/gmic.2.4.16109

The bacterium Clostridium difficile is the leading cause of healthcare associated diarrhoea in the developed world and thus presents a major financial burden. The main virulence factors of C. difficile are two large toxins, A and B. Over the years th... Read More about Both, toxin A and toxin B, are important in Clostridium difficile infection.

Regulation of neurotoxin production and sporulation by a putative agrBD signaling system in proteolytic Clostridium botulinum (2010)
Journal Article
Cooksley, C. M., Davis, I. J., Winzer, K., Chan, W. C., Peck, M. W., & Minton, N. P. (2010). Regulation of neurotoxin production and sporulation by a putative agrBD signaling system in proteolytic Clostridium botulinum. Applied and Environmental Microbiology, 76(13), 4448-4460. https://doi.org/10.1128/AEM.03038-09

A significant number of genome sequences of Clostridium botulinum and related species have now been determined. In silico analysis of these data revealed the presence of two distinct agr loci (agr-1 and agr-2) in all group I strains, each encoding pu... Read More about Regulation of neurotoxin production and sporulation by a putative agrBD signaling system in proteolytic Clostridium botulinum.

Two-component signal transduction system CBO0787/ CBO0786 represses transcription from botulinum neurotoxin promoters in Clostridium botulinum ATCC 3502
Journal Article
Zhang, Z., Korkeala, H., Dahlsten, E., Sahala, E., Heap, J. T., Minton, N. P., & Linström, M. Two-component signal transduction system CBO0787/ CBO0786 represses transcription from botulinum neurotoxin promoters in Clostridium botulinum ATCC 3502. PLoS Pathogens, 9(3), Article e1003252. https://doi.org/10.1371/journal.ppat.1003252

Blocking neurotransmission, botulinum neurotoxin is the most poisonous biological substance known to mankind. Despite its infamy as the scourge of the food industry, the neurotoxin is increasingly used as a pharmaceutical to treat an expanding range... Read More about Two-component signal transduction system CBO0787/ CBO0786 represses transcription from botulinum neurotoxin promoters in Clostridium botulinum ATCC 3502.

Regulation of cel genes of C. cellulolyticum: identification of GlyR2, a transcriptional regulator regulating cel5D gene expression
Journal Article
Fendri, I., Abdou, L., Trotter, V., Dedieu, L., Maamar, H., Minton, N. P., & Tardif, C. Regulation of cel genes of C. cellulolyticum: identification of GlyR2, a transcriptional regulator regulating cel5D gene expression. PLoS ONE, 8(1), Article e44708. https://doi.org/10.1371/journal.pone.0044708

Transcription and expression regulation of some individual cel genes (cel5A, cel5I, cel5D and cel44O) of Clostridium cellulolyticum were investigated. Unlike the cip-cel operon, these genes are transcribed as monocistronic units of transcription, exc... Read More about Regulation of cel genes of C. cellulolyticum: identification of GlyR2, a transcriptional regulator regulating cel5D gene expression.