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

• Published in: Molecular biology reports Vol. 47; no. 3; pp. 1821 - 1834
• Main Authors: Sarkar, Purabi, Stefi, Raju V., Pasupuleti, Mukesh, Paray, Bilal Ahmad, Al-Sadoon, Mohammad K., Arockiaraj, Jesu
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.03.2020; Springer Nature B.V
• Subjects: Adenosylhomocysteinase - chemistry; Animal Anatomy; Animal Biochemistry; Animals; Antioxidants; Antioxidants - chemical synthesis; Antioxidants - chemistry; Antioxidants - pharmacology; Bacterial Proteins - chemistry; Bioinformatics; Biomedical and Life Sciences; Cell adhesion & migration; Cell migration; Cell Movement - drug effects; Cell Proliferation - drug effects; Cell Survival; Cell viability; Chlorocebus aethiops; Cytotoxicity; Fibroblasts; Fibroblasts - cytology; Fibroblasts - drug effects; Fibroblasts - metabolism; Gene expression; Histology; Humans; Intracellular; Leukocytes, Mononuclear - cytology; Leukocytes, Mononuclear - drug effects; Leukocytes, Mononuclear - metabolism; Life Sciences; Morphology; Original; Original Article; Oxidative stress; Peptides; Peptides - chemical synthesis; Peptides - chemistry; Peptides - pharmacology; Reactive Oxygen Species - metabolism; Spirulina - enzymology; Sulfur; Vero Cells; Vitamin E; Wound healing; Wound Healing - drug effects; Adenosyl homocysteinase; Fibroblast cells; Spirulina; Wound healing; Cyanobacteria
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-020-05276-y

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.