High-Throughput Screening and Selection of Pichia pastoris Strains
Clone screening procedures for Pichia pastoris expression strain comparison rely on the availability of a cultivation environment that ensures equal growth and production capabilities for all assessed transformants. As clonal variation in such experiments is caused by diverging numbers and possibly also genomic locations of integrated (linearized) expression constructs, the productivity assessment of a larger number of strains is mandatory for selecting a set of strains for follow-up bioreactor cultivations in order to define the best-producing clone. Microscale cultivation provides the means to reliably compare growth and productivity of a large number of transformants and by that narrows down the amount of selected strains for scaling up.
Key wordsSmall-scale cultivation Deep-well plates High-throughput Protein production Screening
- 2.Hartner FS, Ruth C, Langenegger D, Johnson SN, Hyka P, Lin-Cereghino GP, Lin-Cereghino J, Kovar K, Cregg JM, Glieder A (2008) Promoter library designed for fine-tuned gene expression in Pichia pastoris. Nucleic Acids Res 36(12):e76. https://doi.org/10.1093/nar/gkn369 CrossRefPubMedPubMedCentralGoogle Scholar
- 4.Marsalek L, Gruber C, Altmann F, Aleschko M, Mattanovich D, Gasser B, Puxbaum V (2017) Disruption of genes involved in CORVET complex leads to enhanced secretion of heterologous carboxylesterase only in protease deficient Pichia pastoris. Biotechnol J 12(5). https://doi.org/10.1002/biot.201600584 CrossRefGoogle Scholar
- 8.Camattari A, Goh A, Yee Yip L, Hee Meng Tan A, Wai Ng S, Tran A, Liu G, Liachko I, Dunham MJ, Rancati G (2016) Characterization of a panARS-based episomal vector in the methylotrophic yeast Pichia pastoris for recombinant protein production and synthetic biology applications. Microb Cell Factories 15:139. https://doi.org/10.1186/s12934-016-0540-5 CrossRefGoogle Scholar