Who’s in the team?
Dr Emmanuel Oluwabusola, Dr Federica Casolari, Nicole Cochrane, Professor Marcel Jaspars and Professor Rainer Ebel.
What do you do?
We manage and run the new BBSRC-funded mass spectrometer, a Thermo Orbitrap Tribrid IQ-X liquid chromatograph mass spectrometer (LC-MS). This system is optimised for the analysis and identification of small molecules in complex mixtures. It separates mixtures and breaks molecules into diagnostic fragments (known as MSn). It can also image tissue and sample thin sections to show where certain compounds are distributed in the material. This system can be applied to solve problems in chemistry, biology, medicine and archaeology amongst others.
We are receiving training and are learning to use the hardware and software. Once we are more familiar with the system, we will have information sessions in the different schools to advertise the system’s capabilities and potential use. If you have any project that would benefit from using the system, please contact Marcel Jaspars (m.jaspars@abdn.ac.uk) to discuss the potential application of the system to your research.
The Thermo Orbitrap Tribid IQ-X LC-MS has two ionisation modes (ESI, APCI) three fragmentation modes (CID. HCD and UV-PD) two mass analysers (ion trap, Orbitrap). We were able to secure 5 years’ service and support, as well as a large number of software packages. The AP-MALDI for MS imaging is shared between Chemistry and the Rowett.
Why is it important?
Chemical space is, big, mindbogglingly big. Currently there are over 250,000 human metabolites (small molecules) known, and at least another 250,000 from plants, microorganisms and invertebrates. The number of compounds across all species globally therefore runs into billions of compounds, many in trace amounts, and many with important bioactivity. Identifying and quantifying these compounds is essential for understanding biological processes and disease states, discovering new drugs and engineering enzymes for industrial biotechnology. Less than 5% of all mass spectra can be linked to compound structures. Therefore, an approach that can provide compound identification of each component of a complex mixture with high confidence is essential. Only LC-MSn provides this capacity, when coupled with very high mass resolving power and different fragmentation modes with accurate mass determinations up to very high levels of MSn. Mass spectrometric fragments are used to identify compounds with a high degree of confidence.
What are some of your big projects right now?
Discovery of new antibiotics - With over 670k infections yearly in the EU, antimicrobial resistance has become one of the most pressing health challenges. Microorganisms are acquiring multiple resistance mechanisms against all currently available treatments thus spreading multidrug resistant organisms in the environment. The need for new more selective drugs leads research towards unexplored and extreme environments, such as the deep sea, which is proven to be unique habitat for microorganisms that produce novel antibiotics.
Historically, small molecules from microorganisms have provided a rich source of diverse antibiotic compounds. With the rising resistance to these proven antibiotic classes, alternative sources of antibiotics must be discovered. The ribosomally synthesized and post-translationally modified peptides (RiPPs) have been attracting interest as one such source of untapped potential. The possibility of interpreting their genomes to predict what compounds they make is assisted by having powerful tools like the IQ-X available. This project will focus on identifying these compounds in the extracts of bacteria from extreme environments. The utility of the IQ-X is that we will be able to identify predicted metabolites and verify their structure using fragmentation of selected liquid chromatography peaks of the extract. This has not been possible previously and this capacity will be incredibly useful.
The role of invertebrates in deep ocean environments. Deep-sea invertebrates are important in climate regulation and their potential for bioprospecting. Due to their remote and inaccessible nature, studies on the ecological interactions (competition, symbiosis) and bioprospecting of deep-sea invertebrates remain scarce. The metabolome of the sponge Mycale lingua from various locations is being analysed using LC-MS and MS networking. Unique clusters of metabolites are being isolated and their structures determined. Compound bioactivity is being assessed in a variety of biological and ecological assays. A separate project is studying larger collections of deep-sea invertebrates using metabolomics to investigate the diversity of invertebrates inhabiting abyssal Pacific mining regions and defining their capacity to produce natural products. Critically, this dynamic biodiversity assessment in protected and at-risk regions of the deep sea will enable a fair and informed comparison of the potential impacts of seabed mining compared to terrestrial mining. Given the challenge in collecting materials, often coupled with low sample amounts, structure determination will be achieved mainly using the MSn fragmentation.
Any other key takeaways about your team?
Teamwork is at the core of our group. We enjoy working together, and learning from one another as we have different backgrounds ranging from Natural Product chemistry to molecular biology to plant biology etc. This aids in developing protocols for sample preparation as well as data interpretation. Communication among us forwards the overall understanding of the instrument, both independently and as a team. We are constantly learning from each other, whereby when troubleshooting is required, we tackle issues as a whole. We’ve built our confidence in managing our IQ-X by running a large variety of samples in multiple applications.
What’s the best thing about being part of your team?
Responsibilities are shared; therefore, workload and instrument maintenance are equally distributed.