E. coli strain engineering (pathway engineering)
Escherichia coli is one important producer organism in industrial biotechnology and represents the best studied and understood prokaryotic model organism in microbiology. Genetic modified strains are used constantly to express industrial important proteins, enzymes, or fine chemicals. Metabolic engineering has become more and more attractive to optimize strains to meet commercial objectives by modifying metabolic pathways or even introducing entirely new ones.
Recombineering (Red/ET recombination) permits a defined and rapid access to the chromosomal modification of E. coli by (multiple) gene disruption, deletion and insertion, as well as gene modification by introducing (point) mutations in a seamless manner. Recombineering also allows promotor fine tuning to optimize gene expression in order to remove bottlenecks or to reduce activities of metabolic pathways producing unwanted by-products.
Gen-H’s scientists performed hundreds of E. coli modifications in the last years for academic and industrial customers including very challenging modifications like multiple insertions of expression cassettes into the genome, insertion of large operons or the insertion of point mutations in essential genes.
Gen-H offers its know-how in micro- and molecular biology plus experience in White Biotechnology for the creation of any adjusted E. coli strain.
Our services include:
- Customized design
- Decent timelines and regular progress updates
- Final strain confirmation by PCR and sequencing of modified regions
- If desired: documentation in GMP-like manner
What we offer:
- Modification of K12 strains (e.g. W3110), B-strains (e.g. BL21(DE3)), E. coli W and Nissle1917.
- Single or multiple gene knock-outs
- Single or multiple insertions
- Seamless gene modifications
- Insertion of point mutations in essential genes
- Promoter modulations
- Application of an Escherichia coli triple reporter strain for at-line monitoring of single-cell physiology during L-phenylalanine production; Mahn Dat Hoang, Dieu Thi Doan, Marlen Schmidt, Harald Kranz, Andreas Kremling and Anna-Lena Heins; Eng Life Sci 2022, 1
- Novel Escherichia coli active site dnaE alleles with altered base and sugar selectivity; Alexandra Vaisman, Krystian Łazowski, Martin A. M. Reijns, Erin Walsh, John P. McDonald, Kristiniana C. Moreno, Dominic R. Quiros, Marlen Schmidt, Harald Kranz,Wei Yang, Karolina Makiela-Dzbenska and Roger Woodgate; Molecular Microbiology 2021; 116: 909
- CroSR391, an ortholog of the λ Cro repressor, plays a major role in suppressing polVR391-dependent mutagenesis; John P. McDonald, Dominic R. Quiros, Alexandra Vaisman, Antonio R. Mendez, Jan Reyelt, Marlen Schmidt, Martín Gonzalez and Roger Woodgate; Molecular Microbiology 2021, 116: 877
- Development and characterization of Escherichia coli triple reporter strains for investigation of population heterogeneity in bioprocesses; Anna-Lena Heins, Jan Reyelt, Marlen Schmidt, Harald Kranz and Dirk Weuster-Botz; Microbial Cell fact 2020, 19: 14
- Chromosome and plasmid-borne PlacO3o1 promoters differ in sensitivity to critically low temperatures; Samuel Oliveria, Nadia Goncalves, Vinodh Kandavalli, Leonardo Martins, Ramakanth Neeli-Venkata, Jan Reyelt, Jose Fonseca, Jason Lloyd-Price, Harald Kranz and Andre Ribero; Scientific Reports 2019, 9: 4486
- Investigating the mechanisms of ribonucleotide excision repair in Escherichia coli; Alexandra Vaisman, Jahon McDonald, Stephan Noll, Donald Huston, Gregory Loeb, Myron Goodman and Roger Woodgate; Mutation Research 2014, 761: 21
- Improving safety and performance of the expression host Escherichia coli BL21(DE3); Stephan Noll, Jan Reyelt, Thomas Rysiok, Roland Kellner, Detlef Güssow, Stefan Jäkel, Stefanie Hager and Harald Kranz; Poster Presentation RPP7
- Gezielte Optimierung von Escherichia coli BL21(DE3); Stephan Noll, Jan Reyelt, Thomas Rysiok, Roland Kellner, Detlef Güssow, Stefan Jäkel, Stefanie Hager and Harald Kranz; Biospektrum 2013, 19: 211