Metabolic Engineering of Recombinant Protein Production by Saccharomyces cerevisiae
Doktorsavhandling, 2012

The yeast Saccharomyces cerevisiae is a widely used cell factory for the production of fuels, chemicals, and it also provides a platform for the production of many heterologous proteins of medical or industrial interest. In this thesis, random and rational approaches, such as vector design, host engineering, fermentation analysis, UV Mutation, coupled with high-throughput systems biology techniques (including whole genomic sequencing, microarray analysis and flux analysis) and integrated analysis (Reporter feature technique), were employed to engineer cellular properties more effectively and purposefully to construct cell factories for protein production. We reported that insulin production mainly depends on the expression level of the gene, whereas amylase tends to achieve higher secretion at lower growth conditions in order to reduce ER stress. Moreover, based on large data generated and systems biology tools, we proposed several models to address unknown questions regarding recombinant protein production: i) the futile cycle of protein folding in the ER and the thermodynamic model of non-stoichiometric production of reactive oxygen species explains the oxidative stress that occurred during recombinant protein production, and ii) the final electron acceptor for protein folding and the electron transferring model at anaerobic condition proposed potential electron consuming pathway for protein folding in the ER. Our research provided a deep understanding of the processing of protein secretory pathway, potential targets for future engineering, as well as shed lights for basic cellular metabolisms.

recombinant protein production

genome sequencing

DNA microarray

insulin precursor

growth rate

UV mutation

yeast

anaerobic electron acceptor

secretory pathway

unfolded protein response

metabolic engineering

systems biology

α-amylase

seminar room Euler
Opponent: Prof. Diethard Mattanovich

Författare

Zihe Liu

Chalmers, Kemi- och bioteknik, Livsvetenskaper, Systembiologi

Metabolic engineering of recombinant protein secretion by Saccharomyces cerevisiae

FEMS Yeast Research,; Vol. 12(2012)p. 491-510

Artikel i vetenskaplig tidskrift

Different expression systems for production of recombinant proteins in Saccharomyces cerevisiae

Biotechnology and Bioengineering,; Vol. 109(2012)p. 1259-1268

Artikel i vetenskaplig tidskrift

Imbalance of heterologous protein folding and disulfide bond formation rates yields runaway oxidative stress

BMC Biology,; Vol. 10(2012)p. Art. no 16-

Artikel i vetenskaplig tidskrift

Proteins play crucial roles in cell signaling, immune systems and the cell cycle. Many human proteins have important values or great potentials as biopharmaceutical. Since the first recombinant pharmaceutical was approved for clinical use, recombinant DNA technology and protein engineering have established an efficient tailor-made industry for protein production. Now there are over 300 biopharmaceuticals proteins and antibodies on the market, with more than $100 billion of sales. In addtion, around 240 monoclonal antibody products and 120 recombinant proteins are in clinical trials. The yeast Saccharomyces cerevisiae provides a platform for the production of many heterologous proteins of medical or industrial interest. In this thesis, random and rational approaches, such as vector design, host engineering, fermentation analysis, UV Mutation, coupled with high-throughput systems biology techniques (including whole genomic sequencing, microarray analysis and flux analysis) and integrated analysis (Reporter feature technique), were employed to engineer cellular properties more effectively and purposefully to construct cell factories for protein production. Our research provided a deep understanding of the processing of protein secretory pathway, proposed targets for future engineering, as well as shed lights for basic cellular metabolisms.

Styrkeområden

Livsvetenskaper och teknik

Ämneskategorier

Läkemedelsbioteknik

Annan industriell bioteknik

ISBN

978-91-7385-741-3

Doktorsavhandlingar vid Chalmers tekniska högskola. Ny serie: 3422

seminar room Euler

Opponent: Prof. Diethard Mattanovich