Project Reference: TRG27-BIO-CB2

Project title: The early life of membrane-forming enzymes

Project outline

Understanding how molecular systems became capable of organising themselves into functional cells is a fundamental question in biology. Membranes are central to this process, providing the boundary that defines cells and controlling molecular exchange with their environment. However, little is known about how the enzymes that synthesise and remodel membrane components emerged and diversified. This project will investigate how primitive catalysts could have contributed to membrane formation and function, while developing new technologies for enzyme discovery and engineering.

The project combines the expertise of Dr. Claudia Bonfio in prebiotic chemistry, membrane biophysics and protocell engineering with Prof. Florian Hollfelder’s expertise in high-throughput enzyme discovery, directed evolution and microfluidics. The student will develop an ultrahigh-throughput droplet-based platform to screen large enzyme libraries, including metagenomic sequences from extremophilic organisms and designed variants. Candidates will be tested against plausible prebiotic substrates to identify enzymes capable of synthesising or modifying membrane-forming amphiphiles.

Promising enzymes will then be reconstituted in liposomes to determine how enzymatic activity changes membrane composition and properties. Fluorescence-based assays, zeta potential measurements and Raman spectroscopy will provide complementary information on membrane remodelling.

Finally, directed evolution and phylogenetic approaches will be used to define the functional requirements of membrane-modifying enzymes and generate simplified variants retaining catalytic activity. The project will address a fundamental question concerning the emergence of cellular organisation while establishing a transformative platform for discovering and engineering enzymes. The resulting technology could have applications in synthetic biology, including the development of highly resilient artificial cells and biomolecular compartments.

Principal Supervisor

Dr Claudia Bonfio

Email address: cb2036@cam.ac.uk

Department / Partner Institute: Biochemistry

Website: https://bonfiolab.eu/

BBSRC strategic theme

Transformative technologies