Rapid screening of cellular stress responses in recombinant Pichia pastoris strains using metabolite profiling.
Heterologous protein production in the yeast Pichia pastoris can be limited by biological responses to high expression levels; the unfolded protein response (UPR) is a key determinant of the success of protein production in this organism. Here, we used untargeted NMR metabolic profiling (metabolomics) of a number of different recombinant strains, carried out in a miniaturized format suitable for screening-level experiments. We identified a number of metabolites (from both cell extracts and supernatants) which correlated well with UPR-relevant gene transcripts, and so could be potential biomarkers for future high-throughput screening of large numbers of P. pastoris clones.
Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, London, SW7 2AZ, UK.|Department of Applied Mathematics, Australian National University, Canberra, Australia.
Gregory D
GD
Tredwell
Department of Life Sciences, Faculty of Natural Sciences, Imperial College London, London, SW7 2AZ, UK.
Rochelle
R
Aw
Department of Life Sciences, Faculty of Natural Sciences, Imperial College London, London, SW7 2AZ, UK.|FUJIFILM Diosynth Biotechnologies, Billingham, TS23 1LH, UK.
Bryn
B
Edwards-Jones
Department of Life Sciences, Faculty of Natural Sciences, Imperial College London, London, SW7 2AZ, UK.|Department of Biology and Biochemistry, University of Bath, Bath, BA2 7AY, UK.
David J
DJ
Leak
Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, London, SW7 2AZ, UK. j.bundy@imperial.ac.uk.
Jacob G
0000-0002-1164-8465
JG
Bundy
D057166:High-Throughput Screening Assays; D055432:Metabolomics; D000074041:Microorganisms, Genetically-Modified; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D056811:Unfolded Protein Response / Q000235:genetics
D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins
Bioprocessing; Clonal variation; Metabolic footprinting; Recombinant protein; Unfolded protein response