Increased production of chymosin by glycosylation

Journal of Biotechnology
Hans J M van den BrinkMarianne Harboe

Abstract

Filamentous fungi are well known in the industry as producers of large amounts of extracellular proteins. However, production levels of heterologous proteins are often disappointing low. In this paper it is shown that increasing glycosylation is a powerful strategy for increasing production levels of chymosin in filamentous fungi. Two different concepts based on glycosylation were tested. First, we improved a poorly used N-glycosylation site within the prochymosin molecule. The resulting highly glycosylated chymosin molecule was expressed in Aspergillus niger. It was shown that production of the glycosylated protein was much more efficient, giving a yield increase of more than 100% compared to production of the native chymosin molecule. In an alternative strategy the N-glycosylation site was located outside of the native chymosin molecule, on a linker separating prochymosin from its carrier molecule. Also in this case significantly increased production levels were obtained. This strategy might offer a powerful tool for increasing production levels of other heterologous proteins as well.

References

Oct 1, 1991·Bio/technology·N S Dunn-ColemanR LaCost
Jan 1, 1992·Scandinavian Journal of Clinical and Laboratory Investigation. Supplementum·J AikawaT Beppu
Jan 1, 1992·Methods in Enzymology·P J Punt, C A van den Hondel
Jan 1, 1991·Advances in Experimental Medicine and Biology·J AikawaT Beppu
May 1, 1994·Enzyme and Microbial Technology·N Jenkins, E M Curling
Jan 1, 1997·Applied Microbiology and Biotechnology·R J GoukaC A van den Hondel
Jan 1, 1997·Biotechnology & Genetic Engineering Reviews·P P WardO M Conneely
Oct 9, 2001·Trends in Biochemical Sciences·C M CabralR N Sifers

❮ Previous
Next ❯

Citations

Jun 10, 2010·Applied Microbiology and Biotechnology·André Fleissner, Petra Dersch
Aug 2, 2014·Protein Engineering, Design & Selection : PEDS·Bastiaan A van den BergDick de Ridder
Oct 5, 2011·Biotechnology Advances·Owen P Ward
Oct 9, 2008·Biotechnology Advances·David Lubertozzi, Jay D Keasling
Sep 3, 2010·Journal of the Science of Food and Agriculture·Zhen FengLanwei Zhang
Jan 28, 2012·Journal of Dairy Science·J A VallejoT G Villa
Apr 4, 2015·Protein Expression and Purification·Nan WangDeHu Liu
Jun 8, 2010·Critical Reviews in Biotechnology·A KumarV K Batish
May 21, 2017·Protein Expression and Purification·Wen-Guo LiuMing Lu
Mar 22, 2021·Protein Expression and Purification·Selin Ali HanoğluMehmet Karaaslan

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bioinformatics in Biomedicine

Bioinformatics in biomedicine incorporates computer science, biology, chemistry, medicine, mathematics and statistics. Discover the latest research on bioinformatics in biomedicine here.

Blood Clotting Disorders

Thrombophilia includes conditions with increased tendency for excessive blood clotting. Blood clotting occurs when the body has insufficient amounts of specialized proteins that make blood clot and stop bleeding. Here is the latest research on blood clotting disorders.