Award details

14IBCAT1FEASIBILITY Discovery and development of large/diverse user-friendly panels of novel biocatalysts

Reference	BB/M028496/1
Principal Investigator / Supervisor	Professor Gary Black
Co-Investigators / Co-Supervisors	Professor Justin Perry, Dr MENG ZHANG
Institution	Northumbria University
Department	Fac of Health and Life Sciences
Funding type	Research
Value (£)	82,241
Status	Completed
Type	Research Grant
Start date	01/07/2015
End date	30/06/2016
Duration	12 months

Abstract

The project will involve biochemical assay development. This will include the initial appraisal of the literature and other resources such as the Bergmeyer Methods of Enzymatic Analysis Series, BRENDA, etc, towards the development and optimisation of suitable assay formats for characterisation of the 10 classes of enzymes to be studied. Where possible colorimetric assays will be developed, such as ones based on detection of the NAD(P)H produced from for instance aldehyde dehydrogenases activity, where a red INT-formazan product is formed in the presence of diaphorase activity. Such assays will be very useful for not only characterisation of the new panels to demonstrate the requisite diversity intended, but also to enable advanced screening technology to be developed for subsequent in-house screening by customers of Prozomix. The project will also likely involve the discovery/development of novel activity assays to account for the small number of putative enzymes expected not to exhibit the expected activity. The project will also involve preliminary biochemical characterisation of the 10 classes of enzymes to be studied. This will establish which of the enzymes generated exhibit the expected activity, thus informing subsequent enzyme target selection. These assays will be conducted wherever possible in 96-well high-throughput format, for instance employing a microplate reader, but will also employ lower-throughput assays where necessary, such as those involving LC-MS. Finally, the project will involve detailed biochemical characterisation of the 10 classes of enzymes to be studied. This will include using colorimetric, spectrophotometric, GC(-MS), (chiral) LC(-MS), and/or NMR based methods as appropriate. The data will be used to demonstrate the diversity of novel enzyme specificities discovered, and for the preparation of a series of scientific papers as appropriate.

Summary

Biochemical tests will be developed to facilitate initial screening of members from each of 10 classes of enzyme on a small range of relevant chemical compounds which the enzymes are likely to convert. Those enzymes that are shown to convert one or more such compounds will be characterized in detail. This analysis will employ techniques that involve absorbency of light (spectrophotometry), separation (chromatography) and mass determination (mass spectrometry). Analysis will also involve determining whether certain enzymes can distinguish between non-superimposable mirror images of chemical compounds (i.e. stereoisomers), as this is of critical importance when producing pure compounds for the pharmaceutical industry, for example. It is highly likely that some of the enzymes will not convert the chosen chemical compounds. These enzymes will then be tested against other chemical compounds that will be informed by studying the relevant scientific literature. The results generated will inform the efficient selection of other enzymes, by the industrial partner producing the enzymes, which are complementary to those that are performing interesting conversions. This will increase the total number of enzymes produced that perform interesting conversions, i.e. generate wide-ranging screening panels that will therefore perform a large range of interesting conversions.

Impact Summary

The beneficiaries of this research will be those using biocatalysis. Examples of immediate beneficiaries include the Chemicals, Food Producers and Pharmaceuticals and Biotechnology industries, as well as academia. This research will significantly simplify and drive down the costs of biocatalysis research and development, increasing its uptake by industry and academia. In the longer term, the increasing number of the resulting non-petroleum-based (i.e. bio-based) products produced will contribute significantly to the alleviation of the consumption of resources that harm the environment in terms of biodegradability, toxicity, and pollution resulting in a beneficial effect on the health of the UK population. Importantly, the market for bio-based products is likely to increase significantly in the longer term, and has been identified as a lead market by the European Commission. Therefore it is imperative to wealth creation, and specifically the economic competitiveness of the UK, that the biocatalysis community has access to appropriate enzymes that can contribute to the manufacture of bio-based products.

Committee	Not funded via Committee
Research Topics	Industrial Biotechnology
Research Priority	X – Research Priority information not available
Research Initiative	Industrial Biotechnology Catalyst (IBCAT) [2014-2015]
Funding Scheme	X – not Funded via a specific Funding Scheme

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