Application of repeated aspartate tags to improving extracellular production of Escherichia coli L-asparaginase isozyme II

Enzyme and Microbial Technology
Sun-Ki KimJin-Ho Seo

Abstract

Asparaginase isozyme II from Escherichia coli is a popular enzyme that has been used as a therapeutic agent against acute lymphoblastic leukemia. Here, fusion tag systems consisting of the pelB signal sequence and various lengths of repeated aspartate tags were devised to highly express and to release active asparaginase isozyme II extracellularly in E. coli. Among several constructs, recombinant asparaginase isozyme II fused with the pelB signal sequence and five aspartate tag was secreted efficiently into culture medium at 34.6 U/mg cell of specific activity. By batch fermentation, recombinant E. coli produced 40.8 U/ml asparaginase isozyme II in the medium. In addition, deletion of the gspDE gene reduced extracellular production of asparaginase isozyme II, indicating that secretion of recombinant asparaginase isozyme II was partially ascribed to the recognition by the general secretion machinery. This tag system composed of the pelB signal peptide, and repeated aspartates can be applied to extracellular production of other recombinant proteins.

References

Feb 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·A L SwainA Wlodawer
Apr 5, 2000·Pharmaceutica acta Helvetiae·A L StecherJ Abrahão-Neto
Oct 12, 2000·Current Opinion in Biotechnology·P Cornelis
Apr 20, 2001·Molecular Microbiology·M Sandkvist
May 1, 1964·Archives of Biochemistry and Biophysics·L T MASHBURN, J C WRISTON
Oct 13, 2004·Protein Expression and Purification·Amardeep KhushooK J Mukherjee
Jan 22, 2005·Applied Microbiology and Biotechnology·Amardeep KhushooK J Mukherjee
Mar 15, 2005·Biotechnology Advances·F J M MergulhãoG A Monteiro
Jun 1, 2005·Trends in Biotechnology·David S Waugh
Jun 14, 2006·Bioprocess and Biosystems Engineering·Karolin DreslerWolf-Dieter Deckwer
Jun 20, 2006·Current Opinion in Biotechnology·Dominic Esposito, Deb K Chatterjee
Oct 20, 2006·Proceedings of the National Academy of Sciences of the United States of America·Joseph A SorgOlaf Schneewind
Jul 22, 2008·Journal of Bacteriology·Kelly E RiordanOlaf Schneewind
Sep 11, 2008·Biotechnology and Bioengineering·Hyun-Dong Shin, Rachel R Chen
Jul 15, 2009·Biotechnology Letters·Ye Ni, Rachel Chen
May 11, 2013·Protein Science : a Publication of the Protein Society·Shan FengHaiteng Deng

❮ Previous
Next ❯

Citations

Nov 15, 2018·Current Protein & Peptide Science·Deepankar SharmaAbha Mishra
Apr 14, 2018·Engineering in Life Sciences·Gabriele R M Kleiner-GroteKarl Friehs
Jun 9, 2018·Microorganisms·Vasiliki Paraskevopoulou, Franco H Falcone
Mar 25, 2018·International Journal of Biological Macromolecules·Sarika AgrawalNaveen Kango
Aug 3, 2021·International Journal of Biological Macromolecules·Shu-Ping ZouYu-Guo Zheng

❮ Previous
Next ❯

Related Concepts

Related Feeds

Blood And Marrow Transplantation

The use of hematopoietic stem cell transplantation or blood and marrow transplantation (bmt) is on the increase worldwide. BMT is used to replace damaged or destroyed bone marrow with healthy bone marrow stem cells. Here is the latest research on bone and marrow transplantation.