ESR Project 7
University of Cambridge, Department of Biochemistry, Cambridge, United Kingdom
Directed evolution of enzymes for the synthesis of modified non-natural nucleoside building blocks for oligonucleotide therapeutics
Friederike Nintzel received her bachelor’s degree in Biochemistry from the Ruprecht-Karls-University of Heidelberg, Germany.
During her bachelor’s study, she conducted several research internships: She researched in structural biology with Dr. Christian Biertümpfel at the Max Planck Institute of Biochemistry in Martinsried, Germany. Furthermore, she researched in virology with Prof. Dr. Gülsah Gabriel at the HPI - Leibniz Institute for Experimental Virology in Hamburg, Germany and she performed a research internship in environmental toxicology with Dr. Johan Lundqvist at the VHC Research Centre at SLU Uppsala in Sweden. Additionally, she worked as teaching assistant at the University of Heidelberg.
For her bachelor’s thesis, she researched in phototrophic whole-cell biocatalysis with Prof. Bruno Bühler at the Helmholtz Centre for Environmental Research in Leipzig, Germany. She worked on establishing a synthetic cascade for the conversion of cyclohexane to epsilon-caprolactone in Pseudomonas taiwanensis and Synechocystis sp. PCC 6803.
Friederike pursued a master’s degree in Life Science & Technology with an Annotation in Technology for Sustainable Development at Delft University of Technology in the Netherlands. During her master’s thesis, she researched in enzyme biocatalysis with Prof. Sven Panke at ETH Zürich D-BSSE in collaboration with Prof. Frank Hollmann at TU Delft. Her thesis aimed at immobilisation of peroxygenase rAaeUPO for increased performance in hydrophobic environments and application of the catalyst for synthesis of epoxides in neat reactions (published in Nintzel et al. 2021, Chemical Communications).
Furthermore, being highly enthusiastic about sustainability, she was working at the interface of sustainability, technology, and innovation during an internship at Merck KGaA, Darmstadt, Germany.
Biocatalytic routes to chemical building blocks [e.g. oligonucleotides (ONs) in the treatment of genetic diseases] are emerging as environmentally friendly and sustainable ‘green’ routes for manufacturing and this approach will be developed as part of this multicentre project.
The aim of this ESR PhD project will be the development of efficient biocatalysts. Starting with promiscupus enzymes, strategies for ultrahigh-throughput evolution will be developed, consisting of implementation of droplet-based microfluidic assays, synthesis of gene libraries (via error-prone PCR and transposon-based InDel mutagenesis), improved mutants will be selected and the process of evolution monitored by full-length nanopore sequencing (using UMIC-seq). Alternatively identification of useful enzymes will be attempted by functional metagenomics. Ultimately the utility of new biocatalysts will be tested in collaboration with partner groups.
This project is carried out in strong collaboration with the following groups:
Prof. Roger Strömberg, Karolinska Institutet, Sweden
Dr. Anna Perdrix Rosell, Sixfold Bioscience, UK
Project summary in 3 min
The Hollfelder 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 we extend the mechanistic lessons learned to potential applications in biotechnology, synthetic chemistry and medicine.