Metal-sensing in Salmonella: A model for targeting a network that differentiates metals
Lead Research Organisation:
Durham University
Department Name: Biosciences
Abstract
Metals have been used to control microbes in agriculture, food handling, domestic hygiene, medicine and more broadly as an additive to preserve perishables. The industrial partner, Procter and Gamble, have on-going programmes to develop new metal-based biocides to replace existing preservatives and to increase the efficacy of current metal-based anti-microbials (ensuring compliance with shifting legislative guidelines). Ionophores can help minimise the amount of metal added to products and there is interest in using more subtle combinations of metals and/or chelators. Historically, the exploitation of metals has been empirical but the discovery of natural metal-based antimicrobial mechanisms and of bacterial systems that sense and adapt to metals, presents opportunities to improve these additives through mimicry and subversion respectively.
Metals are implicated in several natural anti-microbial mechanisms. For example hosts and pathogens have evolved to compete for iron. Copper is pumped into the phagolysosomal compartment of macrophages about eight hours post infection most probably as a biocide to drive the Fenton reaction. Neutrophils are thought to starve microbes of zinc and manganese by releasing the metal-binding protein calprotectin. Bacteria (in common with all other types of cells) have evolved elaborate homeostatic mechanisms to balance the buffered intracellular concentrations of various metals within critical thresholds; critical to ensure that vast numbers of metalloproteins acquire their correct metal-cofactors. Central to such metal homeostasis is a set of metal-sensing proteins that detect when the buffered limits have been exceeded. The sensors trigger expression of proteins that restore the correct metal-balance. Having discovered bacterial metal-sensors and identified properties that determine which metals they 'can' sense in vitro, the next step is to investigate how metal-specificity in vivo (the metals the sensors 'do' sense inside cells) is a shared function of a set of metal-sensors. We will begin to model the network of interactions between the different sensor proteins. This programme will explore a fundamental question central to the cell biology of metals, coincidentally providing insight needed to formulate additives that subvert bacterial metal-sensing networks.
Metals are implicated in several natural anti-microbial mechanisms. For example hosts and pathogens have evolved to compete for iron. Copper is pumped into the phagolysosomal compartment of macrophages about eight hours post infection most probably as a biocide to drive the Fenton reaction. Neutrophils are thought to starve microbes of zinc and manganese by releasing the metal-binding protein calprotectin. Bacteria (in common with all other types of cells) have evolved elaborate homeostatic mechanisms to balance the buffered intracellular concentrations of various metals within critical thresholds; critical to ensure that vast numbers of metalloproteins acquire their correct metal-cofactors. Central to such metal homeostasis is a set of metal-sensing proteins that detect when the buffered limits have been exceeded. The sensors trigger expression of proteins that restore the correct metal-balance. Having discovered bacterial metal-sensors and identified properties that determine which metals they 'can' sense in vitro, the next step is to investigate how metal-specificity in vivo (the metals the sensors 'do' sense inside cells) is a shared function of a set of metal-sensors. We will begin to model the network of interactions between the different sensor proteins. This programme will explore a fundamental question central to the cell biology of metals, coincidentally providing insight needed to formulate additives that subvert bacterial metal-sensing networks.
Technical Summary
Five transcriptional regulators that detect metals have been characterised in S. Typhimurium (CueR, GolS, Fur, MntR and Zur). Metal-effectors and operator-promoter targets can be predicted for a further five deduced sensors (hypothetically ZntR, NikR, RcnR, ModE and ArsR) exploiting sequence similarity and gene context. Predicted DNA-binding sites and metal-specificities will be validated by experiment, establishing which metals each sensor responds to under steady state conditions (generations of growth under maximum and minimum permissive concentrations of each metal). One additional CsoR/RcnR homologue plus five further MerR-homologues are also encoded in the genome and will be analysed to discover which ones are metal sensors.
Recombinant proteins will be expressed and purified to determine affinities (of the tightest sensory site in the first instance) for the metals that each sensor detects, affinities for the metals detected by the other sensors and affinities for DNA. This part of the programme will be aided by a linked PhD student, also funded by our industrial partners. In addition to advancing knowledge of the individual components of the set of metal sensors in a micro-organism of relevance to food production and processing, this programme will quantify the abundance of each metal sensor in vivo under each steady state metal condition to enable the generation of integrated mathematical models to describe metal-specificity of metal-sensing. An attractive feature is that some intracellular parameters (the buffered available concentration of metals for example) can be treated as common functions for all of the sensors, assisting the mathematical modelling.
Recombinant proteins will be expressed and purified to determine affinities (of the tightest sensory site in the first instance) for the metals that each sensor detects, affinities for the metals detected by the other sensors and affinities for DNA. This part of the programme will be aided by a linked PhD student, also funded by our industrial partners. In addition to advancing knowledge of the individual components of the set of metal sensors in a micro-organism of relevance to food production and processing, this programme will quantify the abundance of each metal sensor in vivo under each steady state metal condition to enable the generation of integrated mathematical models to describe metal-specificity of metal-sensing. An attractive feature is that some intracellular parameters (the buffered available concentration of metals for example) can be treated as common functions for all of the sensors, assisting the mathematical modelling.
Planned Impact
The logical next step in our on-going (two decades) basic research programme to understand how bacteria sense and discern metals, is to model interactions between the individual metal-sensors. This is expected to discover why some DNA-binding, metal-responsive, transcriptional regulators are competent to respond (allosterically) to certain metals, but fail to do so in vivo. Coincidentally, this research aims to uncover combinations of treatments which subvert the metal-sensory network. For this reason we are extremely fortunate that an industrial sponsor, Procter and Gamble (P&G), has offered to partly finance this fundamental project in the form of an Industrial Partner Award (IPA). In addition the industrial partner is funding a linked PhD student who will work closely with the research associate appointed to this IPA. Moreover, P&G will supply additional expertise to aid the microbial systems modelling, and the latter stages of the project will use their in-house testing facilities.
P&G already exploit metal and/or metal-chelator-based antimicrobial treatments in consumer products for which they hold substantive market shares; as preservatives, in antibacterial cleaning agents and in personal hygiene products. This research programme has the potential to inform the development of more effective anti-microbial treatments exploiting less metal and/or combinations of metals, chelators and other agents. Durham University and Procter and Gamble have a strategic research relationship in which a Master Collaboration Agreement is already in place to cover Intellectual Property, confidentiality, publication and any subsequent commercial exploitation. Within the provision of any overriding terms of the research grant offer letter, these existing agreed terms will be used to govern impact relating to commercial exploitation. Development of this proposal has involved interaction between the University and Industrial partner in the form of visits, conference calls and of course extensive e-mail traffic to review draft documents. Throughout the programme there will be regular meetings and on-going electronic communication. All of these mechanisms will ensure that discoveries with industrial impact are exploited in the swiftest possible manner.
Other forms of impact, the trained personnel and public engagement activities, are noted in the pathways to impact statement. The publications that result from this research may also assist in the development of metal-based antimicrobial treatments for other purposes, for example in the control of plant pathogens (we have support from Syngenta in this area), and more broadly adding to the arsenal of approaches to protect the UK against disease.
P&G already exploit metal and/or metal-chelator-based antimicrobial treatments in consumer products for which they hold substantive market shares; as preservatives, in antibacterial cleaning agents and in personal hygiene products. This research programme has the potential to inform the development of more effective anti-microbial treatments exploiting less metal and/or combinations of metals, chelators and other agents. Durham University and Procter and Gamble have a strategic research relationship in which a Master Collaboration Agreement is already in place to cover Intellectual Property, confidentiality, publication and any subsequent commercial exploitation. Within the provision of any overriding terms of the research grant offer letter, these existing agreed terms will be used to govern impact relating to commercial exploitation. Development of this proposal has involved interaction between the University and Industrial partner in the form of visits, conference calls and of course extensive e-mail traffic to review draft documents. Throughout the programme there will be regular meetings and on-going electronic communication. All of these mechanisms will ensure that discoveries with industrial impact are exploited in the swiftest possible manner.
Other forms of impact, the trained personnel and public engagement activities, are noted in the pathways to impact statement. The publications that result from this research may also assist in the development of metal-based antimicrobial treatments for other purposes, for example in the control of plant pathogens (we have support from Syngenta in this area), and more broadly adding to the arsenal of approaches to protect the UK against disease.
Publications
Osman D
(2019)
Bacterial sensors define intracellular free energies for correct enzyme metalation.
in Nature chemical biology
Young TR
(2021)
Calculating metalation in cells reveals CobW acquires CoII for vitamin B12 biosynthesis while related proteins prefer ZnII.
in Nature communications
Osman D
(2017)
Fine control of metal concentrations is necessary for cells to discern zinc from cobalt.
in Nature communications
Schilter D
(2019)
Finding the right match
in Nature Reviews Chemistry
Foster AW
(2014)
Metal preferences and metallation.
in The Journal of biological chemistry
Osman D
(2015)
Generating a Metal-responsive Transcriptional Regulator to Test What Confers Metal Sensing in Cells.
in The Journal of biological chemistry
Osman D
(2016)
The Effectors and Sensory Sites of Formaldehyde-responsive Regulator FrmR and Metal-sensing Variant.
in The Journal of biological chemistry
Description | The network of metal sensors has been characterised in Salmonella, providing insight into what enables a DNA-binding metal-sensor to detect a specific metal inside a cell. In turn, these findings informed the creation of a DNA-binding metal-sensor from a transcriptional regulator that normally detects formaldehyde (Journal of Biological Chemistry, 2015). By measuring metal-affinities, DNA-affinities of apo- and metalated sensors, plus the number of sensor molecules per cell, it has become possible to combine these parameters to model metal specificity of metal-sensing transcriptional-regulators. This established that the bona fide sensor for each metal is simply the most sensitive in the set of Salmonella sensors for that cognate ion (Nature Communications, 2017). However, these data also established that perfect metal specificity only operates when the buffered metal concentrations are fine tuned to within a narrow range (about one order of magnitude). Outside this narrow window of concentrations, other sensors are liable to mal-respond to the wrong metal, and indeed this was shown to be the case when cells were transiently shocked with elevated metal concentrations (Nature Communications, 2017). The order of mal-responses matched the thermodynamic predictions. These data establish that the metal sensory system of bacteria is vulnerable to subversion by combinations of metal chelants and ionophores, offering an explanation for why metal excess and deficiency appears to be a microbial 'Achillees heel' exploited in nutritional immunity and in a range of antimicrobial treatments. Synergistic combinations of metal chelants have been discovered. Finally, this work has provided major insight into protein metalation. There is a challenge for metalloenzymes to acquire their correct metals because some inorganic elements form more stable complexes with proteins than do others. These preferences can be overcome provided some metals are more available than others. However, while the total amount of cellular metal can be readily measured, the available levels of each metal have been more difficult to define. Metal-sensing transcriptional regulators are tuned to the intracellular availabilities of their cognate ions. In this programme, the standard free energy for metal complex formation to which each sensor, in a set of bacterial metal sensors, is attuned, has been established. This has shown that the less competitive the metal, the less favourable the free energy and hence the greater availability to which the cognate allosteric mechanism is tuned. Crucially, comparing these free energies with values derived from the metal affinities of a metalloprotein reveals the mechanism of correct metalation. This work was published in Nature Chemical Biology in 2019, which was recommended by two members of Faculty 1000 and highlighted by an article in Nature Chemistry, We are working to generate a metalation calculator to allow researchers in academia and industry alike, to determine the metalation state of proteins (and other molecules) inside living cells. For P&G this could include ionophore ingredients with implications for product preservation. For industrial biotechnologists, this could include half of enzymes (the metallo-enzymes) many of which drive critical bioprocesses and bio-transformations. |
Exploitation Route | Refer to impact section. The work also has relevance to the development of new antimicrobials and to the optimisation of metal supply for metalloenzymes in Industrial Biotechnology. By using the free energy values for available metals inside cells (determined under this award) a metalation calculator has now been released (initially as 'Supplementary data 1' in Nature Communications (2021) article, https://www.nature.com/articles/s41467-021-21479-8) to enable others to determine the metalation state of proteins of interest in cells and hence optimise processes exploiting metalloenzymes. This has been put to use in optimising the production of vitamin B12 (https://www.nutritioninsight.com/news/vitamin-b12-calculator-could-reduce-manufacturing-price-amid-rising-vegan-need.html). Also a series of workshops have also been run by the E3B BBSRC NIBB supporting two-way communication about how the calculator can be put to use by others (https://sites.durham.ac.uk/mib-nibb/events/), described within the engagement activities here in Researchfish. |
Sectors | Chemicals Manufacturing including Industrial Biotechology Retail |
Description | This BBSRC IPA research grant with Procter and Gamble has formed the foundation for a series of additional related programmes on microbial-metal-systems in Durham, funded to the tune of between £0.5 and £1 million by Procter and Gamble. These programmes have supported four related PhD students and (at least) three research assistants. The insights from this entire body of research are being exploited by the Industrial partner in relation to the formulation of fast moving consumer goods. Regular meetings between the Durham metals-team and Procter and Gamble staff from the UK and from Cincinnati continued through 2020. This work underpinned a successful application for a phase II BBSRC NIBB and ongoing funded work to develop a metalation calculator for widespread use in bioscience and biotechnology. |
First Year Of Impact | 2019 |
Sector | Manufacturing, including Industrial Biotechology |
Impact Types | Economic |
Description | 12 month JGW GS programme |
Amount | £112,500 (GBP) |
Organisation | Procter & Gamble |
Sector | Private |
Country | United States |
Start | 03/2016 |
End | 03/2017 |
Description | A calculator for metalation inside a cell (Extranet ref: OEFE3B003) |
Amount | £625,780 (GBP) |
Funding ID | BB/V006002/1 |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 09/2021 |
End | 09/2024 |
Description | BBSRC NIBB phase II |
Amount | £1,360,000 (GBP) |
Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
Sector | Public |
Country | United Kingdom |
Start | 03/2019 |
End | 02/2024 |
Description | MB studentship |
Amount | £64,500 (GBP) |
Organisation | Procter & Gamble |
Sector | Private |
Country | United States |
Start | 08/2014 |
End | 09/2018 |
Description | MM studentship |
Amount | £44,000 (GBP) |
Organisation | Procter & Gamble |
Sector | Private |
Country | United States |
Start | 08/2015 |
End | 09/2019 |
Description | MS programme |
Amount | £318,646 (GBP) |
Organisation | Procter & Gamble |
Sector | Private |
Country | United States |
Start | 12/2014 |
End | 12/2017 |
Description | PhD studentship CP |
Amount | £60,000 (GBP) |
Organisation | Procter & Gamble |
Sector | Private |
Country | United States |
Start | 11/2011 |
End | 10/2015 |
Description | RM studentship |
Amount | £99,000 (GBP) |
Organisation | Procter & Gamble |
Sector | Private |
Country | United States |
Start | 09/2012 |
End | 09/2016 |
Description | Research Fellowships - Royal Commission 1851 - to TRY (Extranet ref: OEFE3B001) |
Amount | £250,000 (GBP) |
Organisation | Royal Commission for the Exhibition of 1851 |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 09/2019 |
End | 09/2022 |
Title | Calculating in vivo metalation from the sensitivities of metal-sensors. |
Description | Equations, software and spreadsheets to calculate the sensitivities of metal sensors and in turn to determine metal availability inside a cell. This enables the calculation of metalation inside living cells with implications for engineering a half of the reactions of life. Includes: 1. Excel Spreadsheet (with instructions) to enable calculation of fractional DNA occupancy. 2. MATLAB codes (with instructions), to determine the buffered metal concentration from given value(s) of ?D or ?DM. 3. Supplementary equations and unique Supplementary Note 2 references in support of the above. |
Type Of Material | Technology assay or reagent |
Year Produced | 2019 |
Provided To Others? | Yes |
Impact | Too early |
URL | https://www.nature.com/articles/s41589-018-0211-4.pdf |
Title | Clones encoding proteins of metal homeostasis |
Description | Clones distributed in 2014 include pETInrS, pETNmtR, pETKmtR, pETCucA, pBAD24-ssCucA-GFP-SsrA |
Type Of Material | Biological samples |
Year Produced | 2014 |
Provided To Others? | Yes |
Impact | Supplied to reseachers in Bologna, UMass Boston, Universite de Riems |
Title | Computational method to determine DNA occupancy by metal sensors |
Description | Computational method to determine DNA occupancy by metal sensors |
Type Of Material | Technology assay or reagent |
Year Produced | 2017 |
Provided To Others? | Yes |
Impact | Ongoing industrial collaboration. |
URL | https://www.nature.com/articles/s41467-017-02085-z#Sec23 |
Title | Crystal coordinates of FrmR |
Description | Crystal coordinates of FrmR PDB: 5LCY |
Type Of Material | Database/Collection of data |
Year Produced | 2016 |
Provided To Others? | Yes |
Impact | PDB entry: 5LCY |
URL | http://www.rcsb.org/pdb/explore.do?structureId=5lcy |
Description | Chelation Therapy in the Washing Machine |
Organisation | Procter & Gamble |
Department | Procter & Gamble Technical Centres Limited |
Country | United Kingdom |
Sector | Private |
PI Contribution | We funded and administered a Business Interaction Voucher in a collaboration with Nick Jakubovics, Newcastle University (BIVMiB012) |
Collaborator Contribution | Biofilms are a major problem in all sorts of industrial settings, including bioprocessing facilities. Mechanical biofilm removal is not always possible due to the chemical and physical properties of the contaminated surface and the use of chemical agents is the most appropriate approach for biofilm control. This proposal involves working with an industrial partner (P&G) to develop novel chemical technologies for biofilm removal at low temperatures on a complex surface (laundry). We envisage that successful outcomes can be translated to biofilm control in many different settings including bioprocessing plants. |
Impact | Project on-going |
Start Year | 2016 |
Description | Interaction with Industry partner |
Organisation | Procter & Gamble |
Country | United States |
Sector | Private |
PI Contribution | Regular teleconferences and exchange visits, exchange of data and materials. |
Collaborator Contribution | Regular teleconferences and exchange visits, exchange of data and funding of multiple PhD students and other staff with only an approximate value given above relating to new activities in the most recent ca one year. |
Impact | Joint publication in 2014 and in subsequent years. Other outcomes are mostly subject to NDA's. |
Start Year | 2014 |
Description | Interaction with industrial sponsor |
Organisation | Procter & Gamble |
Country | United States |
Sector | Private |
PI Contribution | Regular teleconference meetings (in excess of 50 over 24 months including all forms of interaction) with industrial collaborator Reciprocal exchange of materials and biologics with industrial collaborator Reciprocal visits with industrial collaborator (associated PhD students and academic staff etc) Analytical services provided for industrial partner and others, and vice versa |
Collaborator Contribution | See above |
Impact | Ongoing and confidential |
Start Year | 2012 |
Title | Metalation calculator as a spreadsheet |
Description | Excel spreadsheet (with instructions) constituting a metalation calculator as "Supplementary Data 1" within the linked publication below |
Type Of Technology | New/Improved Technique/Technology |
Year Produced | 2021 |
Impact | Has enabled the calculation of the metalation state of molecules inside cells in both academic and industrial contexts. |
URL | https://www.nature.com/articles/s41467-021-21479-8 |
Description | Author profile associated with Journal of Biological Chemistry Review. |
Form Of Engagement Activity | A magazine, newsletter or online publication |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Other academic audiences (collaborators, peers etc.) |
Results and Impact | Refer to URL to see the item See next response |
Year(s) Of Engagement Activity | 2014 |
URL | http://www.jbc.org/content/289/41/28095/suppl/DCAuthor_profile |
Description | BBSRC showcase event, Edinburgh |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Professional Practitioners |
Results and Impact | Poster sparked discussion. Further interaction with industrial partner |
Year(s) Of Engagement Activity | 2012 |
Description | EuroBIC bioinorganic chemistry conference |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Disseminated knowledge about the cell biology of metals which sparked questions and discussion. |
Year(s) Of Engagement Activity | 2018 |
URL | https://www.birmingham.ac.uk/facilities/mds-cpd/conferences/eurobic/index.aspx |
Description | FASEB, Lake Tahoe, Trace Metals in Health and Disease |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Dissemination of knowledge about the cell biology of metals which sparked questions and discussion. |
Year(s) Of Engagement Activity | 2018 |
Description | Invited lecture by RA Deenah Osman on Metals in Biology in Hawaii |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Other audiences |
Results and Impact | Invited lecture on the generation of a metals sensor given by RA Deenah Osman. |
Year(s) Of Engagement Activity | 2015 |
Description | Invited lecture, Newcastle University |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Postgraduate students |
Results and Impact | Invited lecture on metals in biology |
Year(s) Of Engagement Activity | 2015 |
Description | Invited lecture, Umea, Sweden |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Public lecture on metals in biology |
Year(s) Of Engagement Activity | 2015 |
Description | Invited seminar by RA (Deenah Osman) at Procter and Gamble, Ohio USA |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Industry/Business |
Results and Impact | Lecture on metals in biology to the Procter and Gamble Mason Business Center in Ohio, USA, given by Deenah Osman and followed by a series of smaller meetings and discussions. |
Year(s) Of Engagement Activity | 2015 |
Description | Invited speaker at the 12th International Biometals web symposium, Biometals 2020. |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Invited speaker in the opening session of an international conference (and also subsequent session chair). |
Year(s) Of Engagement Activity | 2020 |
URL | https://biometals2020.sciencesconf.org/ |
Description | Invited speaker, BBSRC NIBB BioProNET 6th annual science meeting, Manchester |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Industry/Business |
Results and Impact | This aim of the event was to promote collaboration between industry and academia and advertsie the opportunities arising from the E3B BBSRC NIBB. |
Year(s) Of Engagement Activity | 2019 |
URL | http://biopronetuk.org/6th-annual-science-meeting/ |
Description | Invited talk at International Conference on BioInorganic Chemistry, Interlakken, Switzerland |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | The results of our research were described which sparked questions and discussions immediately afterwards and ongoing by e-mail. |
Year(s) Of Engagement Activity | 2019 |
URL | https://www.chem.uzh.ch/dam/jcr:d809e5d0-e81b-42d0-a1c9-175c8e13e958/ICBIC19_ScientificProgram_v5.pd... |
Description | Lecture at the University of Maryland, Baltimore, USA (virtual) |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Invited lecture which increased interest in the subject area. |
Year(s) Of Engagement Activity | 2020 |
Description | Media coverage of publication in Nature Communications |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Industry/Business |
Results and Impact | A press release was generated in support of a publication in Nature Communications as follows: New findings are set to improve the biomanufacturing of B12, a crucial vitamin that is missing from vegan diets, recommended as a supplement by The Vegan Society, but remains prohibitively expensive for many of those who most need it. Vitamin B12 is an essential micronutrient which plays a role in supporting red blood cell production, energy metabolism and nerve function, however it is neither made, nor required by plants. With a record 560,000 people signing up to Veganuary 2021, this important nutrient is in demand and the global transition to low-meat diets means that biomanufacturing will need to increase. However, due to its complex molecular structure, it is currently not feasible to mass-produce via conventional chemical synthesis. Instead, it is the only vitamin which is produced exclusively by bioproduction (culturing bacteria that naturally produce B12). This process remains inefficient and it continues to be expensive for many people who need it - particularly in developing nations. New research, by Dr Tessa Young, of the Department of Biosciences, Durham University, UK, published in Nature Communications, looked into ways of understanding and improving the biosynthesis of B12 by studying how enzymes obtain essential metals. With cobalt, a crucial metal in the B12 production process, Dr Young and the Durham team worked closely with Professor Martin Warren of the University of Kent and the Quadram Institute in Norwich, whose research group engineered E. Coli strains (which don't normally make B12) to synthesise the vitamin. During vitamin B12 biomanufacturing, the vital element, cobalt, is supplied by a metal delivery enzyme. However, ensuring that this enzyme is supplying enough of the right metal, and not becoming clogged-up with the wrong one, remains an obstacle when producing B¬12 on a large scale. To overcome the cobalt bottleneck, Dr Young and the Durham team created a 'metalation calculator' to understand and optimise cobalt supply for B12 to support the manufacture of this essential vitamin. Dr Young said: "By understanding the mechanism that distributes vital metals, it has become possible to produce a calculator which industrial biotechnologists can use to optimise their manufacturing reactions. "The calculator has been tested in the production of vitamin B12 and we hope to see it adopted by biotechnology manufacturers to help foster a more sustainable future." Senior author Professor Nigel Robinson, in the Department of Biosciences, Durham University, said: "About a half of life's reactions are catalysed by metals including iron, copper, zinc, magnesium, manganese, nickel and cobalt. "This paper describes the underlying mechanism that distributes these metals to the reaction centres inside living cells. Industrial Biotechnology manufactures compounds that society needs sustainably, by replacing processes that use fossil fuels with yeast, bacteria or the cells of other organisms as the alternative factories." The ability of the 'metalation calculator' to determine the metal requirements for producing B12 on a large scale shows great promise, not only for the manufacturing of this supplement but also in wider sustainable manufacturing processes using biotechnology. Multiple "tweets" (at least 35 at the time of reporting) relating to these discoveries were also circulated on social media The URL given below is an example of the resulting media coverage. |
Year(s) Of Engagement Activity | 2021 |
URL | https://www.nutritioninsight.com/news/vitamin-b12-calculator-could-reduce-manufacturing-price-amid-r... |
Description | Metal-Related Antimicrobials Workshop |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Industry/Business |
Results and Impact | We organised a workshop to bring together academics and industry to discuss opportunities in collaborative research Led to new collaborations and a volume on "Microbiology of Metals Ions" 2016 volume 70: https://www.elsevier.com/books/microbiology-of-metal-ions/author/978-0-12-812386-7 |
Year(s) Of Engagement Activity | 2015 |
URL | https://www.elsevier.com/books/microbiology-of-metal-ions/author/978-0-12-812386-7 |
Description | Metal-related antimicrobials BBSRC NIBB workshop (November 2015) |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Industry/Business |
Results and Impact | BBSRC NIBB event with representatives from agritechnology business, consumer goods industry, pharmaceutical companies, policy and standards agencies, industrial biotechnology companies to explore the opportunities for metal-related antimicrobials (PI and RA attended). |
Year(s) Of Engagement Activity | 2015 |
Description | Metals in Biology Community Event |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Industry/Business |
Results and Impact | We organised a workshop involving academic and industry to show case funded projects as exemplars to instigate future collaborations. |
Year(s) Of Engagement Activity | 2016 |
Description | Metals in Bioprocessing multiple BBSRC NIBB event (Hexham) |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Industry/Business |
Results and Impact | Interaction with Bio-processing industries. PI and RA both attended and PI gave a presentation. |
Year(s) Of Engagement Activity | 2015 |
Description | NJR Bangalore |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Invited keynote speaker, 6th International conference on Metals in Genetics, Chemical Biology and Therapeutics, Bangalore, India. |
Year(s) Of Engagement Activity | 2016 |
Description | NJR Biometals Dresden Invited Talk |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Invited speaker, Biometals Conference 2016, Dresden, Germany. |
Year(s) Of Engagement Activity | 2016 |
Description | NJR Biophysical Society Invited Lecture |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Invited speaker, British Biophysical Society 2016 Biennial Meeting "Metals in Biology" microsymposium, Liverpool. |
Year(s) Of Engagement Activity | 2016 |
Description | NJR GRC Vermont |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Dissemination of knowldege about the Cell Biology of Metals which sparked questions and discussion. |
Year(s) Of Engagement Activity | 2017 |
URL | https://www.grc.org/cell-biology-of-metals-conference/2017/ |
Description | NJR Queen Mary University of London |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Undergraduate students |
Results and Impact | Delivered the Wills lecture. This sparked questions afterwards followed by requests for information including ICP-MS analyses (for example). |
Year(s) Of Engagement Activity | 2017 |
URL | http://www.sbcs.qmul.ac.uk/research/researchseminars/speciallectureseries/#4 |
Description | NJR San Diego ACS conference |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Invited speaker, American Chemical Society National Meeting Spring 2016, San Diego California. |
Year(s) Of Engagement Activity | 2016 |
Description | NJR TUM Munich |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Delivered lecture and interacted with postgraduate students and staff at a series of meetings. |
Year(s) Of Engagement Activity | 2017 |
Description | National Institutes of Health, Washington DC, presentation |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Disseminated knowledge of the cell biology of metals which sparked questions and discussion. |
Year(s) Of Engagement Activity | 2018 |
Description | Nature Microbiology Community Blog |
Form Of Engagement Activity | Engagement focused website, blog or social media channel |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Invited blog linked to a paper in Nature Communications |
Year(s) Of Engagement Activity | 2017 |
URL | https://naturemicrobiologycommunity.nature.com/users/71254-deenah-osman/posts/24964-sensing-the-diff... |
Description | Organisation of a programme of engagement events involving Industry and Academia related to the exploitation of metal-in-biology expertise in biomaunfacturing, biorecovery and bioenergy. |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Organisation of a series of seminars/workshops to disseminate information about industrial opportunities & challenges, academic expertise & discoveries, and to exemplify projects that connect the two, all within the E3B BBSRC NIBB remit (industrial biotechnology and metals-in-biology). 11 February, 25 February, 4 March and 18 March 2021. The challenge to correctly measure metal affinities of proteins - Many reported metal affinities of proteins are incorrect, often by many orders of magnitude. This one-hour workshop will provide an introduction to some of the common pitfalls and the ways to avoid them. Run by Tessa Young and Nigel Robinson, Durham University. 19 September, 22 September, 29 September 2020 and 16 March 2022. Top tips for writing better manuscripts - a 90-minute interactive seminar designed to help you make the most of your research when publishing a paper, run by a freelance science editor Charlotte Harrison. Aimed at early career researchers, but anyone welcome to attend. 17 February 2021. Probing metalloenzyme catalysis with time-resolved crystallographic and spectroscopic methods at X-ray free-electron lasers; a seminar given by Allen Orville from Diamond Light Source, the UK's national synchrotron science facility. 11 March 2021. Bridging the gap between concept and commercialisation Bob Holt, Centre for Process Innovation Biotechnology. 16 March 2021. UK to get the world's first commercial precious metal bio-refinery from e-waste Ollie Crush and Andy Hanratty, Mint Innovation. 14 April 2021. An Introduction to working with Johnson Matthey Nigel Powell, Johnson Matthey. 17 May 2021. Rare-earth metal responses explored in the genomes of extremophilic red algae Galdieria Seth Davis, University of York. 10 June 2021. Introducing Oxford Biotrans: P450-driven routes to high-value chemicals Matthew Hodges, Oxford Biotrans. 12 October 2021. The London Metallomics Facility Wolfgang Maret and Theodora Stewart. 18 November 2021. Cleaning up biocatalysis with hydrogen: from recycling NADH and flavin cofactors for biotechnology to spin-out of HydRegen and beyond Kylie Vincent and Sarah Cleary, University of Oxford/HydRegen Ltd. 12 January 2022. 'Nuclear Magnetic Resonance' (NMR) Facility Claudia Blindauer and Trent Franks, Warwick University. |
Year(s) Of Engagement Activity | 2021,2022 |
URL | https://mib-nibb.webspace.durham.ac.uk/events/ |
Description | Penn State Summer Symposium in Molecular Biology |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Postgraduate students |
Results and Impact | Disseminated knowledge of the cell biology of metals which sparked questions and discussion. |
Year(s) Of Engagement Activity | 2018 |
Description | Presentation at 'Responsible Innovation: Industrial Biotechnology and Engineering Biology' |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Invited short presentation at 'Responsible Innovation: Industrial Biotechnology and Engineering Biology' Online Event by Carbon Recycling Network & SRBC Nottingham (for BBSRC NIBB) |
Year(s) Of Engagement Activity | 2021 |
URL | https://www.nottingham.ac.uk/iss/events/2020-21/conference-responsible-innovation.aspx |
Description | Presentation to Nobel symposium #168 Visions of bio-inorganic chemistry: metals and the molecules of life, Stockholm |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | The event included 30 pioneer lectures in Bioinorganic Chemistry with a similar number of International observers, leading to discussion about the future of the discipline plus a set of published articles (https://febs.onlinelibrary.wiley.com/toc/18733468/2023/597/1). |
Year(s) Of Engagement Activity | 2022 |
URL | http://doi.org/10.1002/1873-3468.14559 |
Description | RSC Inorganic Biochemistry Discussion Group including industrial uses of metalloenzymes (York, April 2015) |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Industry/Business |
Results and Impact | Discussion of metaloenzymes and their uses (RA attended). |
Year(s) Of Engagement Activity | 2015 |
Description | Tetrapyrroles GRC, Rhode Island |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Disseminated knowledge of the cell biology of metals which sparked questions and discussion. |
Year(s) Of Engagement Activity | 2018 |
URL | https://www.grc.org/chemistry-and-biology-of-tetrapyrroles-conference/2018/ |
Description | The 2020 West Riding Lecture |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Professional Practitioners |
Results and Impact | Named lecture. |
Year(s) Of Engagement Activity | 2020 |