Food Chemistry and Molecular Cancer Biology Lab, Department of Animal Health and Management, Science Campus, Alagappa University, Karaikudi 630 003, India.
National Research Centre for Banana, Thayanur, Tiruchirappalli 620 102, India.
Microb Pathog. 2018 Mar;116:345-350. doi: 10.1016/j.micpath.2018.01.039. Epub 2018 Jan 31.
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), 70 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 IC value of 65.3 ± 0.46 μg mL. 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.
本研究对来自柔鱼的粗乌贼墨的蛋白质组学分析,以研究其抗菌、抗真菌、抗生物膜和细胞毒性特性。为此,使用 SDS-PAGE 将蛋白质分离为条带,进行胶内胰蛋白酶消化,并通过 MALDI-TOF 质谱分析。通过 MASCOT 搜索分析鉴定了 4 条带,分别为类星鼻鱼金属蛋白酶 I (ASMT-I)、70 kDa 神经丝蛋白 (NP)、未知蛋白 LOC106181966 同工型 X1 (UP-Iso-X1)和卵色素结合蛋白 (Oc-BP)。此外,还对获得的粗乌贼蛋白进行了抗菌和抗生物膜活性测试,以分别针对致病性细菌和真菌菌株。此外,还进行了 MTT 测定,以专门解释粗乌贼墨蛋白对 MCF-7 乳腺癌细胞系的细胞毒性能力。研究结果表明,这些蛋白对致病性菌株和乳腺癌细胞系具有剂量依赖性毒性。更重要的是,这些蛋白足以通过光和共聚焦激光扫描显微镜观察来消除生物膜。总之,粗乌贼墨蛋白阻碍了乳腺癌细胞的生长,IC 值为 65.3±0.46μg mL。总之,相信粗乌贼墨中的蛋白质将为未来在阻碍细菌生物膜和癌症方面提供新的见解。