title: Improving heterologous protein secretion at aerobic conditions by activating hypoxia-induced genes in Saccharomyces cerevisiae.
abstract: Oxygen is important for normal aerobic metabolism, as well as for protein production where it is needed for oxidative protein folding. However, several studies have reported that anaerobic conditions seem to be more favorable in terms of recombinant protein production. We were interested in increasing recombinant protein production under aerobic conditions so we focused on Rox1p regulation. Rox1p is a transcriptional regulator, which in oxidative conditions represses genes induced in hypoxia. We deleted ROX1 and studied the effects on the production of recombinant proteins in Saccharomyces cerevisiae. Intriguingly, we found a 100% increase in the recombinant fungal α-amylase yield, as well as productivity. Varied levels of improvements were also observed for the productions of the human insulin precursor and the yeast endogenous enzyme invertase. Based on the genome-wide transcriptional response, we specifically focused on the effect of UPC2 upregulation on protein production and suggested a possible mechanistic explanation.
authors.affiliation: Department of Biology and Biological Engineering, Chalmers University of Technology, SE-41296 Göteborg, Sweden Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE-41296 Göteborg, Sweden.
authors.forename: Lifang
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Biology and Biological Engineering, Chalmers University of Technology, SE-41296 Göteborg, Sweden Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE-41296 Göteborg, Sweden.
authors.forename: Yiming
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Biology and Biological Engineering, Chalmers University of Technology, SE-41296 Göteborg, Sweden Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE-41296 Göteborg, Sweden.
authors.forename: Zihe
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of Biology and Biological Engineering, Chalmers University of Technology, SE-41296 Göteborg, Sweden Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE-41296 Göteborg, Sweden.
authors.forename: Dina
authors.initials: D
authors.lastname: Petranovic
authors.affiliation: Department of Biology and Biological Engineering, Chalmers University of Technology, SE-41296 Göteborg, Sweden Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, SE-41296 Göteborg, Sweden Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, DK-2970 Hørsholm, Denmark nielsenj@chalmers.se.
authors.forename: Jens
authors.initials: J
authors.lastname: Nielsen
mesh_terms: D000332:Aerobiosis; D000693:Anaerobiosis; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D060847:Metabolic Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation
chemical_list: C083348:ROX1 protein, S cerevisiae; D011994:Recombinant Proteins; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; C113958:UPC2 protein, S cerevisiae
keywords: ROX1; UPC2; lipid classes; α–amylase