EastBio CASE: Development of High-Efficiency Landing Pads for Protein Production in P. pastoris

Pichia pastoris, a methylotrophic yeast, is a microbial expression system widely adopted by the biotechnology industry due to its ability to produce recombinant proteins in high yield. While there are well-characterized expression systems, reliable integration and high yield expression of recombinant proteins remains problematic. However, stable and reliable integration of the gene encoding a protein of interest can be facilitated by introducing engineered DNA sequences, called landing pads, into genomic loci typically associated with high gene expression and high protein titre.

In collaboration with our industrial partner Fujifilm Diosynth Biotechnologies (FDBK), we aim to engineer the P. pastoris genome to encode high-efficiency, well-characterised genomic landing pads for heterologous protein expression.

To do that, the student will use transcriptomic and epigenomic data to identify target loci associated with high gene expression. Landing pads, carrying different combinations of promoters and terminators, will be then integrated into the genome using CRISPR and site-specific recombination integration techniques. The engineered strains will be then tested to evaluate the consistency and robustness of protein expression using, as a proof of concept, the the A. niger alpha galactosidase and Human Factor H.

The project builds on existing collaboration between the Prof Giovanni Stracquadanio and Prof Paul Barlow’s group at the School of Biological Sciences and Chemistry.

Applicants should hold a degree in a relevant subject (typically molecular biology, biochemistry, or closely related fields). Previous experience with yeasts, either P. pastoris or S. cerevisiae, is strongly desirable. The student will receive training in microbial protein expression, genome engineering, genomics, transcriptomics and bioinformatics. We expect the successful candidate to build a competitive profile in genome engineering and protein production, which ultimately will support a career in academia or industry.