Performing impactful research requires craftsmanship on the part of the experimenter. Since ZoBio believes that the combination of craftsmanship and leading edge technology will yield impactful insights, we take our responsibility to educate the next generation of scientists seriously. Therefore, ZoBio has participated in the Marie Skłodowska-Curie Innovative Training Network (ITN) FRAGNET whose goal was to train young researchers in the field of Fragment Based Drug Discovery. One of these researchers, Dr. Eleni Makraki, was trained at ZoBio for half a year in NMR spectroscopy.

Eleni’s project focused on a class of industrially important enzymes, the β-glucosidases, which hydrolyse β-1,4-glycosidic bonds. These enzymes are of great use in less environmentally damaging  production of bioethanol from sources such as wood, agricultural residues and dedicated energy crops. However, the β-glucosidase typically is the rate limiting step in conversion and therefore requires very large quantities of enzyme, reducing the economic attractiveness of the process.

The thermostable glycoside hydrolase TrBgl2, is easy to produce and robust and therefore useful for industrial applications. Although engineered versions show enhanced activity, it is desirable to achieve even greater levels. Eleni and the team from University of York used fragment approaches to find a novel activator of TrBgl2.  Even though the crystal structure of the protein was available, efforts to obtain structures of the bound activator were unsuccessful. So, Eleni worked with the ZoBio team to use NMR combined with molecular docking to solve the structure of the complex. One of the conclusions is that covalent linkage of activators would be useful to  generate enzymes that could feasibly be used for industrial applications.

This work has been published in Biochemical Journal (2020) 477 4383-4395.

ZoBio has used all of these techniques in another project to stabilize a preferred conformation of a protein using a specifically designed covalent binder.