Novel extremophilic enzymes for new applications in Healthcare Products (Ref: 4188)

The understanding of cellular communication and microbial biofilm formation is a key interest for Unilever with respect to many of their home and healthcare products which look to target this area for improving hygiene. Enzymes that can interfere with the biofilm formation by specifically hydrolysing the microbial communication lactone molecules are of key importance towards addressing these problems. The potential of combining this approach with other cell wall degrading hydrolases and cleaning chemistries, attempts to create a more holistic approach to multiple modes of action providing the desired outcome. If the multiple mode of intervention is successful it may also help in the prevention of undesired bacterial resistance. This is well aligned to the Clean Future agenda of Unilever and increasing sustainability of products.
Thermostable lactonases have been identified using structural bioinformatics from thermophilic and metagenomes from 'Hot Environments'. The three classes of lactonase enzymes have different structures and different enzymatic mechanisms. The use of these enzymes to break down contaminating biofilms has advantages over the use of antibiotics, and importantly eliminates problems with antibiotic resistance. The Quorum sensing lactonases have been reported to disrupt biofilm formation, the enol lactonases are a new potential target for biofilm disruption and some preliminary information on substrate specificity of these lactonases is available. The Gluconolactonases have been reported to also disrupt biofilm formation.
Recent studies have identified many enzymes useful for improved Healthcare products. Traditionally the majority of industrial enzymes used to date have been obtained from mesophilic organisms, which have a limited stability under harsh industrial conditions including high temperatures, high salt concentrations, extreme pH, and formulation chemicals such as surfactants. Therefore, the discovery of robust enzymes, engineering of more active variants, as well as better understanding of their molecular mechanisms, represent some of the key challenges for the development of future biocatalytic strategies in Industrial Biotechnology. The extremophilic microorganisms represent an attractive source of industrial biocatalysts because they are naturally evolved robust enzymes to function under extreme conditions. In addition, extremophilic enzymes found in one environment (e.g. high temperature) are typically also tolerant to other extreme conditions (e.g. organic solvents, surfactants and other chemical formulation components) making them useful for a variety of applications.

Task 1 - Assessment of novel lactonase and acylases in breakdown of microbial biofilms using quorum sensing lactonases, enol lactonases, gluconolactonases and AHL acylases.

Task 2 - Search Metagenomics DNA databases for further related enzymes. Carry out structurally characterization of selected enzymes by both X-ray crystallography and molecular modelling/ substrate docking. Crystal structures will be used to understand the substrate selectivity and catalytic mechanism of these enzymes and to direct protein engineering experiments to improve their efficacy under industrial conditions.

Task 3 - Evaluation of selected enzyme for application in biofilm disruption. Project placements at Unilever R&D labs within the Material Innovation Factory will involve the use of a bespoke automated microbiological testing platform (for surface adhered microbes), as well as use of fluidic devices and visualisation techniques to observe and measure biofilm development, including gene reporter systems.

Task 4 - All potential lactonases that have positive effects on the formation or disruption of biofilms will be subjected to different types of immobilisation techniques to help develop and broaden their potential applications for home and healthcare products.