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Tham khảo tài liệu 'advances in applied biotechnology part 8', khoa học tự nhiên, công nghệ sinh học phục vụ nhu cầu học tập, nghiên cứu và làm việc hiệu quả | Increasing of Recombinant Protein Production in E. Coli by an Alternative Method to Reduce Acetate 129 rapidly and cannot metabolise the delivered carbon source fast enough Andersen von Meyenburg 1980 Holms 1986 . It is generally observed that even low concentrations of acetate can hamper growth and obstruct the production of recombinant proteins Jensen Carlsen 1990 Nakano et al. 1997 . Many efforts have been made to overcome these hurdles and hence to increase recombinant protein production in E. coli or to express more complex proteins in this host. These engineering attempts are summarized in Fig. 1. Fig. 1. Overview of different engineering approaches to increase recombinant protein production in Escherichia coli The primarily used approach to produce recombinant proteins is to clone the gene of interest on a multi-copy plasmid under the control of a strong promoter in order to achieve high transcription rates and hence high recombinant protein concentrations. However problems such as metabolic burden segregational instability misfolding and proteolytic breakdown or aggregation in inclusion bodies and difficulties in controlling gene expression are usually associated with multi-copy plasmids and the use of strong promoters Noack et al. 1981 Parsell Sauer 1989 Bentley et al. 1990 Dong et al. 1995 Kurland Dong 1996 Gill et al. 2000 Hoffmann Rinas 2004 Ventura Villaverde 2006 . Most engineering strategies to tackle these problems focus on prevention of misfolding neutralisation of increased protease activity or stress response Chou 2007 . An elaborated review of these efforts is given in Waegeman Soetaert 2011 . Two post-translational modifications which are pivotal for the stability and activity of many more complex eukaryotic proteins are disulfide bonds and glycosylation. The former is being facilitated in E. coli by secreting the recombinant protein into the more oxidizing perisplasmic space using the Sec or Tat secretion system by altering the redox state of
