Defense involvement of piscidin from striped murrel Channa striatus and its peptides CsRG12 and CsLC11 involvement in an antimicrobial and antibiofilm activity.

In this study, we have evaluated bioinformatics characterization and antimicrobial role of two piscidin (Pi) peptide identified from the established transcriptome of striped murrel Channa striatus (Cs). The identified CsPi cDNA contains 256 nucleotides encode a protein with 70 amino acids in length which has two antimicrobial peptides and named CsRG12 and CsLC11. The gene expression analysis with various immune stimulants indicated an induced expression pattern of CsPi. Antibiogram showed that CsRG12 and CsLC11 was active against Staphylococcus aureus ATCC 33592, a major multi-drug resistant (MDR) bacterial pathogen and Bacillus cereus ATCC 2106. The minimum inhibitory concentration (MIC) and antibiofilm assays were conducted to observe the activity of pathogenic bacteria with these derived antimicrobial peptides. Flow cytometry analysis noticed that the CsRG12 and CsLC11 disrupt the membrane formation of S. aureus and B. cereus, which was further assured by scanning electron microscopic (SEM) images that bleb formation leads to disruption around the bacterial membrane. Overall, it is reported that CsPi is involved in innate immunity as the gene expression plays a remarkable role in up and down regulation during infection. In addition, the involvement of peptides in antibiofilm formation and bacterial membrane disruption support its immune character. This study leads to a possibility for the development of therapeutics in aquaculture biotechnology.

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 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.