Suppressing posttranslational gluconoylation of heterologous proteins by metabolic engineering of Escherichia coli

Applied and Environmental Microbiology
Juan C AonPramatesh S Patel

Abstract

Minimization of chemical modifications during the production of proteins for pharmaceutical and medical applications is of fundamental and practical importance. The gluconoylation of heterologously expressed protein which is observed in Escherichia coli BL21(DE3) constitutes one such undesired posttranslational modification. We postulated that formation of gluconoylated/phosphogluconoylated products of heterologous proteins is caused by the accumulation of 6-phosphogluconolactone due to the absence of phosphogluconolactonase (PGL) in the pentose phosphate pathway. The results obtained demonstrate that overexpression of a heterologous PGL in BL21(DE3) suppresses the formation of the gluconoylated adducts in the therapeutic proteins studied. When this E. coli strain was grown in high-cell-density fed-batch cultures with an extra copy of the pgl gene, we found that the biomass yield and specific productivity of a heterologous 18-kDa protein increased simultaneously by 50 and 60%, respectively. The higher level of PGL expression allowed E. coli strain BL21(DE3) to satisfy the extra demand for precursors, as well as the energy requirements, in order to replicate plasmid DNA and express heterologous genes, as metabolic flux analysis ...Continue Reading

References

Jul 25, 1997·Clinica Chimica Acta; International Journal of Clinical Chemistry·R M LindsayJ D Baird
Nov 5, 1997·Protein Expression and Purification·P J BertiA C Storer
Sep 2, 1999·Biochemical and Biophysical Research Communications·Z YanP A McDonell
Oct 6, 1999·Current Opinion in Biotechnology·F Baneyx
Apr 4, 2000·Biotechnology and Bioengineering·Santosh B NoronhaJoseph Shiloach
May 2, 2001·Molecular and Cellular Biochemistry·M BeránekV Palicka
Jan 14, 2003·Nature Medicine·Sean B JosephPeter Tontonoz
Jan 29, 2003·Proceedings of the National Academy of Sciences of the United States of America·Veronique HannaertFred R Opperdoes
Oct 4, 2003·The Journal of Biological Chemistry·Roumyana MironovaIvan Ivanov
Jul 29, 2004·The Journal of Biological Chemistry·Qin YanJoan Weliky Conaway
Jul 23, 2005·Annals of the New York Academy of Sciences·Alan W Stitt

Citations

Apr 21, 2009·Journal of Bacteriology·Jennifer I HandfordTracy Palmer
Jan 15, 2014·The Journal of General Physiology·D Brent HallingRichard W Aldrich
Oct 17, 2013·Biotechnology Letters·Quinn Lu, Juan C Aon
Jan 22, 2013·Nature Chemical Biology·Carole L LinsterAndrew D Hanson
May 18, 2012·Plant Molecular Biology·Zerihun A DemissieSoheil S Mahmoud
Feb 24, 2016·Journal of Molecular Biology·Kyle TrainorElizabeth M Meiering
Nov 10, 2009·Proteomics·Marta P Pereira MoraisJean M H van den Elsen
May 27, 2015·Protein Expression and Purification·Aaron M MarcellaAdam W Barb
Aug 1, 2015·ELife·Shi-Hui DongWilfred A van der Donk
Jan 20, 2016·Chemical Communications : Chem Comm·M Robert J ValléeBenjamin G Davis
Feb 2, 2018·Journal of the American Society for Mass Spectrometry·Mowei ZhouLjiljana Paša-Tolić
Dec 23, 2011·Chembiochem : a European Journal of Chemical Biology·Sebastian MeierJens Ø Duus
Sep 18, 2009·Rapid Communications in Mass Spectrometry : RCM·Peiran LiuReb J Russell
Mar 16, 2019·Scientific Reports·Anne-Marie C AnderssonMark Howarth
Apr 17, 2019·Nature Communications·Irsyad N A Khairil AnuarMark Howarth
Feb 10, 2019·Journal of Biomolecular NMR·David SchweidaMario Schubert
Dec 6, 2020·The Journal of Biological Chemistry·Aml A AlnaasJefferson D Knight

Related Concepts

D-glucono-delta-lactone hydrolase
6-phosphogluconolactone
Carboxylic Ester Hydrolases
High Pressure Liquid Chromatography Procedure
Alkalescens-Dispar Group
Fluorometry
Gluconates
Post-Translational Protein Processing
Proteins, Recombinant DNA
Mass Spectrometry

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