Lipase immobilization for microscopic investigation of enzyme activity

Water contamination caused by human and industrial activities is a significant global concern. One of the most prominent pollutants is oily wastewater, severely impacting groundwater and drinking water quality.

Biocatalysts, such as Thermomyces lanuginosus lipase (TLL), are used in many household detergents to remove lipids effectively. However, their limited solubility and reusability can increase running costs and hinder large-scale applications.

To address this issue, we plan to immobilize TLL on solid supports and determine the appropriate surface density of enzymes, conformational changes during immobilization, and the effect of support on its kinetic properties.

Our goal is to develop innovative methods for nanoscale imaging of enzymes using transmission electron microscopy (TEM) to gain insights into the structural aspects of immobilization.

By investigating the interaction of the immobilized enzyme with the support nanostructure and the lipid substrate, we expect to identify the attributes that maximize the biocatalytic reaction rate.

We will explore different immobilization methods on various nanostructures and investigate how TLL interacts with the immobilization matrices at the nanoscale level.

Additionally, we will apply water vapor atmosphere in the TEM to explore in situ the dynamic switching between the active and idle state of the single enzyme molecules in real-time by their conformational changes.

We are aiming to obtain groundbreaking results and a paradigm shift, based on the in situ kinetic studies of lipase-catalyzed chemical reactions which can become a gateway into the quantum mechanical world of molecular science.