Phd on topic: Novel Pichia pastoris platform strains for protein production

Research interest and scientific background: The Glieder group at the Graz University of Technology established crucial gene expression technologies for protein production and engineering of enzymes which are difficult to express and the balanced expression of such enzymes for whole cell catalyzed biosynthetic routes. Genome engineering and editing are applied to generate new host strains for protein production (1-3).

Affiliation: The student will work at the Institute of Molecular Biotechnology at the Graz University of Technology. Cooperation is intended especially with Margit Winkler, and international industrial and academic partners. This project is connected to the “Christian Doppler Laboratory for Pichia pastoris Host and Vector Development”.

Hypothesis and objective: The project aims to investigate the effect of systematic gene inactivation and activation studies on the efficiency of protein production by Pichia pastoris.

Experimental approaches: Novel CRISPR/CAS9 strategies in combination with classical recombination engineering will be applied to generate new host strains with re-engineered genomes. The new strains will be studied and characterized by small scale cultivation in 0,5 ml scale up to controlled cultivation in 5L bioreactors.

The student will strongly interact with other research groups of the industrial partners of the CD lab and international partners at academic labs for the optimization of cultivation technologies, as well as, for bio-computational analyses of the engineered strain employing NextGen sequencing technologies. The student will be embedded

In the doctoral training network of NAWI Graz.


  1. Vogl T, Kickenweiz T, Pitzer J, Sturmberger L, Weninger A, Biggs BW, Koehler EM, Baumschlager A, Fischer J, Hyden P., Wagner M, Baumann M, Borth N, Geier M, Ajikumar PK, Glieder A. Engineered bidirectional promoters enable rapid multi-gene co-expression optimization. Nature Communications, volume 9, Article number: 3589 (2018)
  2. Weninger A, Fischer JE, Raschmanová H, Kniely C, Vogl T, Glieder A. Expanding the CRISPR/Cas9 toolkit for Pichia pastoris with efficient donor integration and alternative resistance markers. J Cell Biochem. 2018 Apr;119(4):3183-3198. doi: 10.1002/jcb.26474.
  3. Sturmberger L, Chappell T, Geier M, Krainer F, Day KJ, Vide U, Trstenjak S, Schiefer A, Richardson T, Soriaga L, Darnhofer B, Birner-Gruenberger R, Glick BS, Tolstorukov I, Cregg J, Madden K, Glieder A. Refined Pichia pastoris reference genome sequence. J Biotechnol. 2016 Oct 10;235:121-31. doi: 10.1016/j.jbiotec.2016.04.023.., PMID: 27084056.