Enzyme discovery projects often start with literature or screening evidence suggesting an enzyme activity associated with a microbial strain. The discovery project can then involve a range of activities from genome sequencing, transcriptional analysis and bioinformatics-guided approaches to finding enzyme sequences for expression and analysis.
We can design new assays to find and assess the activity of enzyme preparations or modify and adapt previously developed assays. Ultimately, our aim is to identify the enzyme sequence encoding the activity of interest, typically following expression in a microbial strain such as Escherichia coli or Pichia pastoris.
Enzyme engineering describes the process whereby the amino acid sequence of an enzyme is altered to improve or adjust its catalytic capability. This can involve rationally guided, targeted changes to an enzyme sequence, supported by bioinformatics and/or methods which rely on iterative selection of properties, such as with directed evolution.
Isomerase has expertise and technologies which enable the manipulation of simple and complex enzymes. From the manipulation of modular multi-domain polyketide synthases to the directed evolution of individual cytochrome P450 oxidases, we work across a wide range of target systems. We also have proprietary bioinformatic software (Evoselect) that improves the selection of active sequences based on evolutionary data.
Following engineering and optimisation of the production process, the microbial strain can then be used in a cellular format to directly produce the product of interest at scale (such as the production of a novel from an engineered non-ribosomal peptide synthetase) or to produce and isolate the enzyme to enable a cell-free biocatalytic process.
Example enzyme discovery and engineering projects
Example projects that make use of our enzyme services include:
- Enzyme Identification or discovery – Genome sequencing of a microbial strain or strains with literature evidence of an enzyme activity. Bioinformatic analysis to generate a shortlist of candidate enzyme sequences. Expression and assay of the resulting enzymes to confirm the presence of desired biocatalytic activity.
- Engineering of the active site of a biocatalytic enzyme - based on in silico structural prediction and/or medium to high throughput assessment of mutant enzymes to look for changes in the product profile.
- Domain engineering - for example, a complex modular enzyme, to bring in enzyme domains to alter the product of a multi-enzyme synthetase to produce a different polyketide or peptide.
- Hybrid enzymes - for example merging two enzymes with identified domains encoding different functions, such as glycosyltransferase substrate recognition domains.
What are the fundamentals of Enzyme Engineering?
There are many techniques that can be used for enzyme engineering including directed evolution, rational design, and computational modelling. Enzyme engineering has the potential to revolutionise many industries by enabling the development of more efficient and sustainable processes, which can lead to cost and/or environmental benefits.
Enzyme engineering can enable the development of new products and services such as new more efficient routes to APIs, bioremediation technologies for environmental cleanup and enzyme-based sensors for detecting contaminants in food and water.
At Isomerase, our enzyme services can be used to enhance the performance of enzymes for specific applications, such as improving the rate or specificity of reaction steps in the production of pharmaceuticals or enabling enzymes to work under specific conditions, such as high temperatures or extreme pH levels. This can result in a more efficient process resulting in cost savings and improved product quality.
Why is Enzyme Engineering important?
Enzyme engineering can enable the optimization of enzymes for specific applications, resulting in improved performance, efficiency, and sustainability.
It can also enable the development of more sustainable processes that help to reduce waste and energy consumption thereby improving the environmental impact of the final product
Not only this but enzyme engineering can also provide a competitive advantage in enabling the development of proprietary enzyme variants that are optimised for specific applications, thereby providing a product with differentiated properties.
Why choose Isomerase?
Isomerase offers a range of innovative enzyme services and platform technologies across bioinformatics, enzyme discovery, enzyme engineering and enzyme manufacture. We have proprietary software, strains and genome sequences that can support enzyme development projects.
We aim to support partners with all aspects of process discovery and development from concept through to scaled manufacture.
Contact us for more information about our enzyme services and to request a quote from us today.