ABSTRACT
The present study deals with the proteomics analysis of crude squid ink isolated from Sepia esculenta for their antibacterial, antifungal, antibiofilm and cytotoxic properties. To achieve this, SDS-PAGE was used to separate proteins as bands, In-gel trypsin digested and analyzed by MALDI-TOF mass spectrometry. A total of 4 bands were identified by MASCOT search analysis namely astacin-like squid metalloprotease type I (ASMT-I), 70â¯kDa neurofilament protein (NP), uncharacterized protein LOC106181966 isoform X1 (UP-Iso-X1) and Ommochrome-binding protein (Oc-BP). Further, the obtained crude squid proteins were subjected to antimicrobial and antibiofilm activities against pathogenic bacterial and fungal strains respectively. Further, MTT assay was also carried out to deliberately explain the cytotoxic ability of crude squid ink protein against MCF-7 breast cancer cell lines. The results from the study revealed that, the proteins are shown to be toxic against pathogenic strains and breast cancer cell lines in a dose-dependent manner. More importantly, the proteins are well enough to eradicate biofilms substantiated by light and confocal laser scanning microscopic observations. Altogether, the crude squid ink proteins hampered the growth of breast cancer cells with an IC50 value of 65.3⯱â¯0.46⯵gâ¯mL-1. In conclusion, it is believed that the proteins from crude squid ink will provide new insights in hampering bacterial biofilms and cancer in near future.
Subject(s)
Anti-Infective Agents/chemistry , Anti-Infective Agents/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Pigments, Biological/chemistry , Proteome/analysis , Sepia/metabolism , Animals , Anti-Infective Agents/isolation & purification , Antineoplastic Agents/isolation & purification , Bacteria/drug effects , Biofilms/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Fungi/drug effects , Humans , MCF-7 Cells , Pigments, Biological/isolation & purification , Proteomics , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
UNLABELLED: Common effluent treatment plant (CETP) for tannery effluent, is the combination of physical, chemical and biological treatment to facilitate the degradation of industrial waste water. Obviously, the biomass which survives in this extreme environment may have the ability to utilize the effluent as the sole carbon source for its survival. The ultimate aim of the present investigation is to expose the microbial diversity in each stage of the CETP through the culture dependent way. Bacterial diversity in the effluent were analysed through 16S rRNA gene. The community study revealed the dominance of firmicutes and the dominant genus was Bacillus sp, with variable species diversity. Notably, Putative Bacillus sp, B. firmus and B. licheniformis were observed in all stages of treatment. The dominant residents were analysed by BProm and TF site scan to prove their uniqueness. This species richness indicates the capability of liveliness in treatment plant and whose can be exploited for treating the effluent by using modern molecular approach. ABBREVIATIONS: CETP - Common Effluent Treatment Plant, PTIET - Pallavaram Tanners Industrial Effluent Treatment Company Ltd.
