The impact of phosphate scarcity on pharmaceutical protein production in S. cerevisiae: linking transcriptomic insights to phenotypic responses

Microbial Cell Factories

Research

Lisbeth Olsson¹  Eva Akke Palmqvist²  Ali Kazemi Seresht³ 

[1] Department of Chemical and Biological Engineering, Industrial Biotechnology, Chalmers University of Technology, 41296, Gothenburg, Sweden;Protein Expression, Novo Nordisk A/S, Novo Nordisk Park 1, 2760, Måløv, Denmark;Protein Expression, Novo Nordisk A/S, Novo Nordisk Park 1, 2760, Måløv, Denmark;Department of Chemical and Biological Engineering, Industrial Biotechnology, Chalmers University of Technology, 41296, Gothenburg, Sweden;

关键词: Phosphate regulation;    heterologous protein production;    chemostat cultivations;    human insulin;    secretory flux;    TPI1;   

DOI  :  10.1186/1475-2859-10-104

 received in 2011-09-13, accepted in 2011-12-07,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundThe adaptation of unicellular organisms like Saccharomyces cerevisiae to alternating nutrient availability is of great fundamental and applied interest, as understanding how eukaryotic cells respond to variations in their nutrient supply has implications spanning from physiological insights to biotechnological applications.ResultsThe impact of a step-wise restricted supply of phosphate on the physiological state of S. cerevisiae cells producing human Insulin was studied. The focus was to determine the changes within the global gene expression of cells being cultured to an industrially relevant high cell density of 33 g/l cell dry weight and under six distinct phosphate concentrations, ranging from 33 mM (unlimited) to 2.6 mM (limited). An increased flux through the secretory pathway, being induced by the PHO circuit during low Pi supplementation, proved to enhance the secretory production of the heterologous protein. The re-distribution of the carbon flux from biomass formation towards increased glycerol production under low phosphate led to increased transcript levels of the insulin gene, which was under the regulation of the TPI1 promoter.ConclusionsOur study underlines the dynamic character of adaptive responses of cells towards a change in their nutrient access. The gradual decrease of the phosphate supply resulted in a step-wise modulated phenotypic response, thereby alternating the specific productivity and the secretory flux. Our work emphasizes the importance of reduced phosphate supply for improved secretory production of heterologous proteins.

【 授权许可】

Unknown   
© Seresht et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.