Show simple item record

dc.creatorAl-Madboly, LA
dc.creatorAli, SM
dc.creatorFakharany, EME
dc.creatorRagab, AE
dc.creatorKhedr, EG
dc.creatorElokely, KM
dc.date.accessioned2020-12-10T16:57:23Z
dc.date.available2020-12-10T16:57:23Z
dc.date.issued2020-02-27
dc.identifier.issn2296-4185
dc.identifier.issn2296-4185
dc.identifier.doihttp://dx.doi.org/10.34944/dspace/4248
dc.identifier.otherLC7JU (isidoc)
dc.identifier.other32181246 (pubmed)
dc.identifier.urihttp://hdl.handle.net/20.500.12613/4266
dc.description.abstract© Copyright © 2020 Al-Madboly, Ali, Fakharany, Ragab, Khedr and Elokely. More attention has been recently directed toward glutathione peroxidase and s-transferase enzymes because of the great importance they hold with respect to their applications in the pharmaceutical field. This work was conducted to optimize the production and characterize glutathione peroxidase and glutathione s-transferase produced by Lactobacillus plantarum KU720558 using Plackett-Burman and Box-Behnken statistical designs. To assess the impact of the culture conditions on the microbial production of the enzymes, colorimetric methods were used. Following data analysis, the optimum conditions that enhanced the s-transferase yield were the De Man-Rogosa-Sharp (MRS) broth as a basal medium supplemented with 0.1% urea, 0.075% H2O2, 0.5% 1-butanol, 0.0125% amino acids, and 0.05% SDS at pH 6.0 and anaerobically incubated for 24 h at 40°C. The optimum s-transferase specific activity was 1789.5 U/mg of protein, which was ~12 times the activity of the basal medium. For peroxidase, the best medium composition was 0.17% urea, 0.025% bile salt, 7.5% Na Cl, 0.05% H2O2, 0.05% SDS, and 2% ethanol added to the MRS broth at pH 6.0 and anaerobically incubated for 24 h at 40°C. Furthermore, the optimum peroxidase specific activity was 612.5 U/mg of protein, indicating that its activity was 22 times higher than the activity recorded in the basal medium. After SDS-PAGE analysis, GST and GPx showed a single protein band of 25 and 18 kDa, respectively. They were able to retain their activities at an optimal temperature of 40°C for an hour and pH range 4–7. The 3D model of both enzymes was constructed showing helical structures, sheet and loops. Protein cavities were also detected to define druggable sites. GST model had two large pockets; 185Å3 and 71 Å3 with druggability score 0.5–0.8. For GPx, the pockets were relatively smaller, 71 Å3 and 32 Å3 with druggability score (0.65–0.66). Therefore, the present study showed that the consortium components as well as the stress-based conditions used could express both enzymes with enhanced productivity, recommending their application based on the obtained results.
dc.format.extent78-
dc.language.isoeng
dc.relation.haspartFrontiers in Bioengineering and Biotechnology
dc.relation.isreferencedbyFrontiers Media SA
dc.rightsCC BY
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectLactobacillus plantarum
dc.subjectglutathione s-transferase
dc.subjectglutathione-peroxidase
dc.subjectenzymatic activity
dc.subjectpurification
dc.titleStress-Based Production, and Characterization of Glutathione Peroxidase and Glutathione S-Transferase Enzymes From Lactobacillus plantarum
dc.typeArticle
dc.type.genreJournal Article
dc.relation.doi10.3389/fbioe.2020.00078
dc.ada.noteFor Americans with Disabilities Act (ADA) accommodation, including help with reading this content, please contact scholarshare@temple.edu
dc.creator.orcidElokely, Khaled M.|0000-0002-2394-021X
dc.date.updated2020-12-10T16:57:19Z
refterms.dateFOA2020-12-10T16:57:24Z


Files in this item

Thumbnail
Name:
Stress-Based Production, and ...
Size:
1.300Mb
Format:
PDF

This item appears in the following Collection(s)

Show simple item record

CC BY
Except where otherwise noted, this item's license is described as CC BY