Version 1
: Received: 21 June 2024 / Approved: 24 June 2024 / Online: 24 June 2024 (08:44:01 CEST)
How to cite:
Gad, A. A. M.; Gora-Sochacka, A.; Sirko, A. Recombinant C-terminal Catalytic Domain of Rat L-gulono Lactone Oxidase Produced in Bacterial Cells Is Enzymatically Active. Preprints2024, 2024061617. https://doi.org/10.20944/preprints202406.1617.v1
Gad, A. A. M.; Gora-Sochacka, A.; Sirko, A. Recombinant C-terminal Catalytic Domain of Rat L-gulono Lactone Oxidase Produced in Bacterial Cells Is Enzymatically Active. Preprints 2024, 2024061617. https://doi.org/10.20944/preprints202406.1617.v1
Gad, A. A. M.; Gora-Sochacka, A.; Sirko, A. Recombinant C-terminal Catalytic Domain of Rat L-gulono Lactone Oxidase Produced in Bacterial Cells Is Enzymatically Active. Preprints2024, 2024061617. https://doi.org/10.20944/preprints202406.1617.v1
APA Style
Gad, A. A. M., Gora-Sochacka, A., & Sirko, A. (2024). Recombinant C-terminal Catalytic Domain of Rat L-gulono Lactone Oxidase Produced in Bacterial Cells Is Enzymatically Active. Preprints. https://doi.org/10.20944/preprints202406.1617.v1
Chicago/Turabian Style
Gad, A. A. M., Anna Gora-Sochacka and Agnieszka Sirko. 2024 "Recombinant C-terminal Catalytic Domain of Rat L-gulono Lactone Oxidase Produced in Bacterial Cells Is Enzymatically Active" Preprints. https://doi.org/10.20944/preprints202406.1617.v1
Abstract
L-gulonolactone oxidase enzyme (GULO) catalyzes the last step of L-ascorbic acid (vitamin C) biosynthesis. This enzymatic activity is lost in primates. The full-length rat GULO has been previously produced in plants and demonstrated to be active. In this study, we compare activity of two variants of GULO produced in Escheriachia coli cells, full-length rat GULO (fGULO) and its C-terminal catalytic domain (cGULO). The expression and purification of the recombinant proteins were optimised and their biological activity was confirmed by two methods, the GULO activity assay in the protein extracts and the ‘in-gel’ staining for GULO activity. Both variants of recombinant GULO were biologically active in both assays. However, cGULO is more promising than fGULO for ascorbic acid production because it is more efficiently produced by bacteria.
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
