Acetylene hydratase enzymes are produced by bacteria to convert acetylene, C2H2, into acetaldehyde, H3C-C(O)H under low-energy, low-pollution conditions (ambient temperature, atmospheric pressure and using water as the solvent). The ability to harness this reaction in industrial biotechnology would provide “greener” ways to synthesise important chemical building blocks from acetylene, which is a cheap industrial-chemistry feedstock. This has significant potential in fuel, plastic and drug production. This project will identify the gene(s) which encode the oxygen-tolerant acetylene hydratase in the bacteria Rhodococcus opacus. This is the crucial first step in initiating a collaborative biology-chemistry project to understand the mechanism of action of acetylene hydratases, with a view to their development in enzyme-catalysed industrial synthesis.
This project was not directly successful (it did not lead to the scientific results anticipated) because an unexpected problem was found when following published methodology (see below). However, by successfully seeding the collaboration between Parkin and Bruce, York hosted a BBSRC NIBB Metals in Biology funded academic and industrial on Rhodococcus (2/11/15) which has been followed up by a publication in Adjacent Government and has seeded a collaboration between Alison and her co-chair Colin Murrell at UEA, leading to an External Speaker invitation for Alison and upcoming summer experiments on different Rhodococcus strains with known sequences to try and further the scientific search for a new family of enzymes, the Mo acetylene hydratases. Additionally, Parkin has also been able to begin collaborating on related hydratase enzymes with Gideon Grogan and an application has been submitted to BBSRC.
A 2015 application for a University EPSRC funded strategic studentship was unsuccessful. A 2013 application for a GlaxoSmithKline UK Industrial CASE studentship in Chemistry (starting Autumn 2014) was unsuccessful.
Objective 1: Initial culturing experiments (project student work, Parkin lab)
Develop methods for growing colonies of Rhodococcus opacus bacteria with and without acetylene, and differentiate (via assays) between conditions which do/do not produce microbes capable of catalysing the reaction between acetylene and water
Successful: The project work described in this application was successfully carried out by a Biochemistry project student (Alec Banner) in Parkin's lab for the academic year 2014/15. The assay method which was to be followed from the literature turned out to display a fundamental flaw, in that it was not a sole measure for acetylene production.
Objective 2: RNA extraction (project student work, Bruce lab)
Conduct RNA extraction experiments to fingerprint how protein expression changes as a function of microbial acetylene hydration activity
Unsuccessful: We did not get to this stage of the project because of an inability to be able to quantify enzyme activity (see above).
Objective 3: Sequencing (Technology facility)
Use sequencing to isolate the gene(s) which encode Rhodococcus opacus acetylene hydratase
Successful: these results are reported in Alec Banner's Biochemistry Project Report and are beig used to support future work.