Understanding enzyme surface kinetics via high content screening methodology

Enzymes are one of the most important molecular tools for laundry and dishwashing detergents, driving soil removal, dingy cleaning, fabric appearance/softness, whiteness maintenance, anti-malodour, and soil release benefits. Industry enzymes play increasingly important roles in making biotechnology more effective and substantiable. When used in laundry detergents or dishwashing products, enzymes can provide improved performance with reduced temperatures, lower water consumption and mitigate environmental impacts. This project is part of a wider effort by P&G and academic partners to develop a new generation of highly effective industry enzymes by investigating several aspects of enzyme performance at the molecular level. Specifically, this project, will be investigating the factors that determine how, and how strongly, enzymes become in contact with the substrates they act upon. This is important because these 'adsorption/desorption kinetics' play a critical role in determining how effective enzymes are in 'real life' consumer products. Several methods are currently used to study enzyme adsorption/desorption kinetics and correlate the results to observed wash performance. For example, Surface Plasmon Resonance (SPR), Dual Polarization Interferometry (DPI) and Quartz Crystal Microbalance with Dissipation (QCM-D). However, these methods are unable to measure enzyme adsorption/desorption kinetics on the surfaces relevant to 'real life' detergent applications since they all use chips with model surfaces that have questionable relevance to textiles, ceramics, steel and other surfaces relevant to detergent enzymes. Hence, this project will develop and apply several new approaches and techniques for studying adsorption/desorption kinetics. Whilst many different enzymes are involved in effective laundry detergents and dishwashing products, carbohydrate active enzymes (CAZymes) will be focussed on initially. Following which lipase, amylase, protease and phosphodiesterase will be looked at.
Project objectives:
Develop a high content screening approach to study enzyme adsorption/desorption kinetics on fabrics and hard surfaces.
Developing a high content screening methodology will first involve a suitable fluorescent enzyme tagging approach to be developed. Previous proof of concept work identified that lipase can be fluorescently tagged with DyLight 594 where it was also found that there is no significant difference in enzyme activity with and without fluorescent tag. A flow cell array will be developed to simultaneously compare the adsorption/desorption properties of enzymes on clean and soiled fabrics following bioimaging using time lapse confocal microscopy.
Application of the methodology to construct a performance model linked to adsorption/desorption kinetics, intrinsic activity/substrate specificity and in-wash stability.
High content screening with high content imaging and data modelling will provide a state-of-the-art approach that utilises imaging of floe cells to measure enzyme surface adsorption/desorption onto relevant surfaces in real time presence of detergent. This will enable P&G to screen multiple conditions in an automated and repeatable manner, replicating the laundry and dishwashing wash cycles enabling to i) enhance the performance of current enzymes via formula modification with desire surface kinetics and ii) engineer new enzymes with desired surface kinetics with our enzyme supplier to achieve the benefits in Cold & Quick wash that can't be achieved with current enzymes.