Cargando…

Antioxidant molecular mechanism of adenosyl homocysteinase from cyanobacteria and its wound healing process in fibroblast cells

An antioxidant molecule namely, adenosyl homocysteinase (AHc) was identified from the earlier constructed transcriptome database of Spirulina, where it was cultured in a sulphur deprived condition. From the AHc protein, a small peptide NL13 was identified using bioinformatics tools and was predicted...

Descripción completa

Detalles Bibliográficos
Autores principales: Sarkar, Purabi, Stefi, Raju V., Pasupuleti, Mukesh, Paray, Bilal Ahmad, Al-Sadoon, Mohammad K., Arockiaraj, Jesu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7223595/
https://www.ncbi.nlm.nih.gov/pubmed/31989428
http://dx.doi.org/10.1007/s11033-020-05276-y
_version_ 1783533769835151360
author Sarkar, Purabi
Stefi, Raju V.
Pasupuleti, Mukesh
Paray, Bilal Ahmad
Al-Sadoon, Mohammad K.
Arockiaraj, Jesu
author_facet Sarkar, Purabi
Stefi, Raju V.
Pasupuleti, Mukesh
Paray, Bilal Ahmad
Al-Sadoon, Mohammad K.
Arockiaraj, Jesu
author_sort Sarkar, Purabi
collection PubMed
description An antioxidant molecule namely, adenosyl homocysteinase (AHc) was identified from the earlier constructed transcriptome database of Spirulina, where it was cultured in a sulphur deprived condition. From the AHc protein, a small peptide NL13 was identified using bioinformatics tools and was predicted to have antioxidant property. Further, the peptide was synthesised and its antioxidant mechanism was addressed at molecular level. NL13 was subjected to various antioxidant assays including DPPH assay, HARS assay, SARS Assay, NO assay and ABTS assay, where NL13 exhibited significant (P < 0.05) potential antioxidant activity compared to its antioxidant control, Trolox. Cytotoxicity was performed on Human whole blood and the cell viability was performed on VERO fibroblast cells. In both assays, it was found that NL13 did not exhibit any cytotoxic effect towards the cells. Further, the intracellular ROS was performed on Multimode reader followed by imaging on fluorescence microscope which showed scavenging activity even at lower concentration of NL13 (31.2 µM). An effective wound healing property of NL13 on VERO cells was confirmed by analysing the cell migration rate at two different time intervals (24 and 48 h). Overall, the study shows that NL13 peptide scavenges the intracellular oxidative stress. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s11033-020-05276-y) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-7223595
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Springer Netherlands
record_format MEDLINE/PubMed
spelling pubmed-72235952020-05-15 Antioxidant molecular mechanism of adenosyl homocysteinase from cyanobacteria and its wound healing process in fibroblast cells Sarkar, Purabi Stefi, Raju V. Pasupuleti, Mukesh Paray, Bilal Ahmad Al-Sadoon, Mohammad K. Arockiaraj, Jesu Mol Biol Rep Original Article An antioxidant molecule namely, adenosyl homocysteinase (AHc) was identified from the earlier constructed transcriptome database of Spirulina, where it was cultured in a sulphur deprived condition. From the AHc protein, a small peptide NL13 was identified using bioinformatics tools and was predicted to have antioxidant property. Further, the peptide was synthesised and its antioxidant mechanism was addressed at molecular level. NL13 was subjected to various antioxidant assays including DPPH assay, HARS assay, SARS Assay, NO assay and ABTS assay, where NL13 exhibited significant (P < 0.05) potential antioxidant activity compared to its antioxidant control, Trolox. Cytotoxicity was performed on Human whole blood and the cell viability was performed on VERO fibroblast cells. In both assays, it was found that NL13 did not exhibit any cytotoxic effect towards the cells. Further, the intracellular ROS was performed on Multimode reader followed by imaging on fluorescence microscope which showed scavenging activity even at lower concentration of NL13 (31.2 µM). An effective wound healing property of NL13 on VERO cells was confirmed by analysing the cell migration rate at two different time intervals (24 and 48 h). Overall, the study shows that NL13 peptide scavenges the intracellular oxidative stress. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s11033-020-05276-y) contains supplementary material, which is available to authorized users. Springer Netherlands 2020-01-27 2020 /pmc/articles/PMC7223595/ /pubmed/31989428 http://dx.doi.org/10.1007/s11033-020-05276-y Text en © Springer Nature B.V. 2020 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Original Article
Sarkar, Purabi
Stefi, Raju V.
Pasupuleti, Mukesh
Paray, Bilal Ahmad
Al-Sadoon, Mohammad K.
Arockiaraj, Jesu
Antioxidant molecular mechanism of adenosyl homocysteinase from cyanobacteria and its wound healing process in fibroblast cells
title Antioxidant molecular mechanism of adenosyl homocysteinase from cyanobacteria and its wound healing process in fibroblast cells
title_full Antioxidant molecular mechanism of adenosyl homocysteinase from cyanobacteria and its wound healing process in fibroblast cells
title_fullStr Antioxidant molecular mechanism of adenosyl homocysteinase from cyanobacteria and its wound healing process in fibroblast cells
title_full_unstemmed Antioxidant molecular mechanism of adenosyl homocysteinase from cyanobacteria and its wound healing process in fibroblast cells
title_short Antioxidant molecular mechanism of adenosyl homocysteinase from cyanobacteria and its wound healing process in fibroblast cells
title_sort antioxidant molecular mechanism of adenosyl homocysteinase from cyanobacteria and its wound healing process in fibroblast cells
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7223595/
https://www.ncbi.nlm.nih.gov/pubmed/31989428
http://dx.doi.org/10.1007/s11033-020-05276-y
work_keys_str_mv AT sarkarpurabi antioxidantmolecularmechanismofadenosylhomocysteinasefromcyanobacteriaanditswoundhealingprocessinfibroblastcells
AT stefirajuv antioxidantmolecularmechanismofadenosylhomocysteinasefromcyanobacteriaanditswoundhealingprocessinfibroblastcells
AT pasupuletimukesh antioxidantmolecularmechanismofadenosylhomocysteinasefromcyanobacteriaanditswoundhealingprocessinfibroblastcells
AT paraybilalahmad antioxidantmolecularmechanismofadenosylhomocysteinasefromcyanobacteriaanditswoundhealingprocessinfibroblastcells
AT alsadoonmohammadk antioxidantmolecularmechanismofadenosylhomocysteinasefromcyanobacteriaanditswoundhealingprocessinfibroblastcells
AT arockiarajjesu antioxidantmolecularmechanismofadenosylhomocysteinasefromcyanobacteriaanditswoundhealingprocessinfibroblastcells