Biography
Florian Hollfelder was educated at the Technical University of Berlin (Diplom-Chemiker) and Cambridge University (MPhil). After a formative stay at Stanford (with Dan Herschlag) on free-energy relationships in enzymes) he joined Tony Kirby’s group at the Chemistry Department of Cambridge University working on enzyme models and physical-organic chemistry. During his PhD he also collaborated with Dan Tawfik (on the mechanism and evaluation of model enzymes such as catalytic antibodies). His postdoctoral work at Harvard Medical School (with Chris T. Walsh) was concerned with the biosynthesis and action of the natural antibiotic microcin B17.
In 2001 he returned to Cambridge to start his own research group in the Biochemistry Department, where he is Professor in Chemical and Synthetic Biology. The group’s research centres around quantitative and mechanistic questions at the chemistry/biology interface, involving low- and high-throughput approaches.
Florian is holder of an ERC Advanced Grant, Director of Studies and Graduate Mentor at Trinity Hall. He is coordinator of several EU-funded trans-national collaborative initiatives, e.g. the present H2020 research training networks MMBio ('Molecular Tools for Nucleic Acid Manipulation for Biological Intervention') and ES-Cat ('Directed Protein Evolution for Synthetic Biology and Biocatalysis'). These initiatives follow structured graduate training programmes developed in previous Marie-Curie networks on directed evolution of functional proteins (ENDIRPRO, ENEFP), on protein-protein interactions (ProSA) and on the chemical biology of phosphates (PhosChemRec).
Research
Nature has evolved the most amazing functional biomolecules and we want to know how they work. Enzymes are the all-purpose catalysts that make the Chemistry of Life run smoothly and efficiently, under the mildest, ‘greenest’ conditions – and protein binders are involved in governing many biological processes.
This group is interested in gaining a fundamental understanding of the principles responsible for molecular recognition processes in chemistry and biology. We probe whether these principles enable us to describe, manipulate and ultimately make functional molecules. Using an eclectic mix of techniques (including kinetic measurements, organic synthesis, biophysical analysis and microengineering) we extend the mechanistic lessons learned to potential applications in biotechnology, chemistry and medicine. In particular we are excited about the potential of pico- to nanolitre mircrodroplets to provide a quantitative high-throughput system that provides insight not only into one reaction at a time, but - by way of massive multiplexing - lets us study millions of samples in one go.
Publications
• Gielen F, Hours R, et al. Ultrahigh-throughput-directed enzyme evolution by absorbance-activated droplet sorting (AADS). Proc. Natl Acad. Sci USA 2016, 113(47), E7383-E7389. Read online
• Colin PY, et al. Ultrahigh-throughput discovery of promiscuous enzymes by picodroplet functional metagenomics. Nature Communications 2015, 6, 10008. Read online
• Colin PY, Zinchenko A, Hollfelder F. Enzyme engineering in biomimetic compartments. Curr Opin Struct Biol 2015, 33, 42-51. Read online
• Shafee T, Gatti-Lafranconi P, Minter R, Hollfelder F. Handicap-Recover Evolution Leads to a Chemically Versatile, Nucleophile-Permissive Protease. Chembiochem 2015, 16(13), 1866–1869. Read online
• Fischlechner M, Schaerli Y, Mohamed MF, Patil S, Abell C & Hollfelder F. Microfluidic Gel-Shell Beads: Evolution of Catalysts Caged in Biomimetic Compartments. Nature Chemistry 2014, 6(9):791-6. Read online
• Diamante, L.; Gatti-Lafranconi, P.; Schaerli, Y.; Hollfelder, F., In vitro affinity screening of protein and peptide binders by megavalent bead surface display. Protein Eng Des Sel 2013, 26(10):713-24. Read online
• Mohamed, M.F. & Hollfelder, F. Efficient, crosswise catalytic promiscuity among enzymes that catalyze phosphoryl transfer. Biochim Biophys Acta 2013, 1834, 417-2. Read online
• Kwok, A., Eggimann, G.A., Reymond, J.L., Darbre, T. & Hollfelder, F. Peptide dendrimer/lipid hybrid systems are efficient DNA transfection reagents: structure-activity relationships highlight the role of charge distribution across dendrimer generations. ACS Nano 2013, 7, 4668-82. Read online
• Gielen, F.; van Vliet, L.; Koprowski, B. T.; Devenish, S. R.; Fischlechner, M.; Edel, J. B.; Niu, X.; Demello, A. J.; Hollfelder, F., A Fully Unsupervised Compartment-on-Demand Platform for Precise Nanoliter Assays of Time-Dependent Steady-State Enzyme Kinetics and Inhibition. Anal Chem 2013, 85(9):4761-9. Read online
• Kintses, B.; Hein, C.; Mohamed, M. F.; Fischlechner, M.; Courtois, F.; Lainé, C.; Hollfelder, F. Chemistry & Biology 2012, Picoliter cell lysate assays in microfluidic droplet compartments for directed enzyme evolution, 19(8):1001-9. Read online
• Villiers, B. R.; Hollfelder, F., Directed evolution of a gatekeeper domain in nonribosomal peptide synthesis. Chemistry & Biology 2011,18(10):1290-9. Read online
• van Loo, B.; Jonas, S.; Babtie, A. C.; Benjdia, A.; Berteau, O.; Hyvönen, M.; Hollfelder, F., An efficient, multiply promiscuous hydrolase in the alkaline phosphatase superfamily. Proc. Natl Acad. Sci USA 2010, 107 (7), 2740-5. Read online
• Golicnik, M.; Olguin, L. F.; Feng, G.; Baxter, N. J.; Waltho, J. P.; Williams, N. H.; Hollfelder, F., Kinetic analysis of beta-phosphoglucomutase and its inhibition by magnesium fluoride. J Am Chem Soc 2009,131 (4), 1575-88. Read online
• Shim, J. U.; Olguin, L. F.; Whyte, G.; Scott, D.; Babtie, A.; Abell, C.; Huck, W. T.; Hollfelder, F., Simultaneous determination of gene expression and enzymatic activity in individual bacterial cells in microdroplet compartments. J Am Chem Soc 2009, 131 (42), 15251-6. Read online
