Appl Microbiol Biotechnol DOI 10.1007/s00253-014-5739-y
APPLIED GENETICS AND MOLECULAR BIOTECHNOLOGY
Enhancing recombinant protein production with an Escherichia coli host strain lacking insertion sequences Myung Keun Park & Sang Hee Lee & Kyung Seok Yang & Suk-Chae Jung & Jun Hyoung Lee & Sun Chang Kim
Received: 26 February 2014 / Revised: 27 March 2014 / Accepted: 29 March 2014 # Springer-Verlag Berlin Heidelberg 2014
Abstract The genomic stability and integrity of host strains are critical for the production of recombinant proteins in biotechnology. Bacterial genomes contain numerous jumping genetic elements, the insertion sequences (ISs) that cause a variety of genetic rearrangements, resulting in adverse effects such as genome and recombinant plasmid instability. To minimize the harmful effects of ISs on the expression of recombinant proteins in Escherichia coli, we developed an IS-free, minimized E. coli strain (MS56) in which about 23 % of the genome, including all ISs and many unnecessary genes, was removed. Here, we compared the expression profiles of recombinant proteins such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and bone morphogenetic protein-2 (BMP2) in MG1655 and MS56. Hopping of ISs (IS1, IS3, or IS5) into the TRAIL and BMP2 genes occurred at the rate of ~10−8/gene/h in MG1655 whereas such events were not observed in MS56. Even though IS hopping occurred very rarely (10−8/gene/h), cells containing the IS-inserted TRAIL and BMP2 plasmids became dominant (~52 % of the total population) 28 h after fermentation began due to their growth advantage over cells containing intact plasmids, significantly reducing recombinant protein production in batch fermentation. Our findings clearly indicate that IS hopping is detrimental to the industrial production of recombinant proteins,
Electronic supplementary material The online version of this article (doi:10.1007/s00253-014-5739-y) contains supplementary material, which is available to authorized users. M. K. Park : S. H. Lee : K. S. Yang : S.
APPLIED GENETICS AND MOLECULAR BIOTECHNOLOGY
Enhancing recombinant protein production with an Escherichia coli host strain lacking insertion sequences Myung Keun Park & Sang Hee Lee & Kyung Seok Yang & Suk-Chae Jung & Jun Hyoung Lee & Sun Chang Kim
Received: 26 February 2014 / Revised: 27 March 2014 / Accepted: 29 March 2014 # Springer-Verlag Berlin Heidelberg 2014
Abstract The genomic stability and integrity of host strains are critical for the production of recombinant proteins in biotechnology. Bacterial genomes contain numerous jumping genetic elements, the insertion sequences (ISs) that cause a variety of genetic rearrangements, resulting in adverse effects such as genome and recombinant plasmid instability. To minimize the harmful effects of ISs on the expression of recombinant proteins in Escherichia coli, we developed an IS-free, minimized E. coli strain (MS56) in which about 23 % of the genome, including all ISs and many unnecessary genes, was removed. Here, we compared the expression profiles of recombinant proteins such as tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and bone morphogenetic protein-2 (BMP2) in MG1655 and MS56. Hopping of ISs (IS1, IS3, or IS5) into the TRAIL and BMP2 genes occurred at the rate of ~10−8/gene/h in MG1655 whereas such events were not observed in MS56. Even though IS hopping occurred very rarely (10−8/gene/h), cells containing the IS-inserted TRAIL and BMP2 plasmids became dominant (~52 % of the total population) 28 h after fermentation began due to their growth advantage over cells containing intact plasmids, significantly reducing recombinant protein production in batch fermentation. Our findings clearly indicate that IS hopping is detrimental to the industrial production of recombinant proteins,
Electronic supplementary material The online version of this article (doi:10.1007/s00253-014-5739-y) contains supplementary material, which is available to authorized users. M. K. Park : S. H. Lee : K. S. Yang : S.
