Making Smarter and Sweeter E. coli
E. coli are great cell factories- they grow fast and on really simple food. This makes them very attractive for biomanufacturing. Not only this, they are most understood organisms from a genetic level with an array of wonderful models to better understand metabolism and predict responses to genetic or environmental perturbation. Their relatively simple cell chassis makes them great for making lots of products from fuels to proteins but how about making more complex molecules more akin to eukaryotic cells?
We have run several projects on generating E. coli cell lines capable of N-type glycosylation. We use a variety of metabolic engineering strategies to increase glycosylation efficiency, including inverse metabolic engineering using genome modification tools, as well as high throughput proteomics approaches with statistical metabolic mapping. To help use along the way we have also develop new quantitative mass spectrometry methods to quantify and characterise glycosylation simultaneously.
We have also contributed to other E. coli based projects.
Please see our publications!
N-Glycosylation in E. coli
Producing a glycosylating Escherichia coli cell factory: The placement of the bacterial oligosaccharyltransferase pglB onto the genome
Strutton B, Jaffé SR, Pandhal J, Wright PC. (2018) Biochemical and Biophysical Research Communications, 495(1): 686-692.
Generation of recombinant N-linked glycoproteins in E. coli
Strutton B, Jaffé SR, Wright PC., Pandhal J, (2017) Methods Molecular Biology, Vol. 1586.
Inverse metabolic engineering for enhanced glycoprotein production in Escherichia coli
Jaffé SR, Strutton B, Pandhal J, Wright PC. (2015) Methods Mol Biol 1321:17-35
Escherichia coli as a glycoprotein production host: recent developments and challenges
Jaffé SR, Strutton B, Levarski Z, Pandhal J, Wright PC. (2015) Current Opinion Biotechnology 30:205-210
Structural and functional characterization of NanU, a novel high-affinity sialic acid-inducible binding protein of oral and gut-dwelling Bacteroidetes species
Phansopa C, Roy S, Rafferty JB, Douglas CW, Pandhal J, Wright PC, Kelly DJ, Stafford GP. (2014) Journal of Biochemistry 458(3):499-511.
Inverse metabolic engineering to improve Escherichia coli as an N-glycosylation host
Pandhal J, Woodruff L.B., Jaffe S, Desai P, Ow S.Y., Noirel J, Gill R.T., Wright P.C. (2013) Biotechnology and Bioengineering 110(9):2482-93
Genome-scale identification and characterization of ethanol tolerance genes in Escherichia coli.
Woodruff L.B., Pandhal J., Ow S.Y., Karimpour-Fard A, Weiss S.J., Wright P.C., Gill R.T. (2013) Metabolic engineering 15:124-33
Systematic metabolic engineering for improvement of glycosylation efficiency in Escherichia coli
Pandhal J., Desai P., Walpole C., Doroudi L., Malyshev D., Wright PC. (2012) Biochemical and Biophysical Research Communications 419(3):472-476
Improving N-glycosylation efficiency in E. coli using high-throughput proteomics, metabolic network analysis and selective reaction monitoring
Pandhal J., Ow S.Y., Noirel J., Wright P.C. (2011) Biotechnology and Bioengineering 108(4):902-912.
N-Linked glycoengineering for human therapeutic proteins in bacteria
Pandhal J., Wright P.C. (2010) Biotechnology letters 32(9):1189-1198.