S Neelima, M V Anju, K Archana, V V Anooja, P P Athira, M R Revathy, M Dhaneesha, A Muneer, T P Sajeevan, S Muhammed Musthafa, I S Bright Singh, S Muraleedharan Nair, Rosamma Philip
{"title":"一种新的来自橄榄云杉的 I 型甲壳素异构体及其抗菌潜力:作为作用模式的膜去极化、破坏和活性氧物种诱导","authors":"S Neelima, M V Anju, K Archana, V V Anooja, P P Athira, M R Revathy, M Dhaneesha, A Muneer, T P Sajeevan, S Muhammed Musthafa, I S Bright Singh, S Muraleedharan Nair, Rosamma Philip","doi":"10.1007/s12602-025-10469-7","DOIUrl":null,"url":null,"abstract":"<p><p>Availability of novel antimicrobial agents is an urgent necessity to combat the growing threat posed by multidrug-resistant bacteria, prompting exploration of marine antimicrobial peptides (AMPs) as potential solutions. Crustacean AMPs are equitably diverse in terms of structure and function, making them consistent templates for novel antimicrobials. From Scylla olivacea gill transcriptome cDNA, a putative Crustin AMP sequence of 333 nucleotides, encoding a 111 amino acid Crustin Type-I isoform was identified. The mature peptide encoding region was cloned and recombinantly expressed in E. coli using Luria Bertani (LB) broth, yielding approximately 0.9 mg/L peptide. This cationic (+6.25) peptide with amphipathic properties (34% hydrophobicity) exhibited antibacterial effects against Gram-positive and Gram-negative strains, with MIC 16 μM against Vibrio spp. The identified modes of actions included disruption of bacterial membranes, membrane potential dissipation, and induction of ROS. Scanning electron microscopy (SEM) analysis revealed bacterial lysis and structural damage. Being non-toxic to mammalian cells (CHO-K1) and non-haemolytic, So-Crustin qualifies to be safe for therapeutic applications. It was quite stable under different physio-chemical/biological conditions, including temperature, pH, NaCl concentrations and proteases like trypsin and proteinase K. This study emphasizes So-Crustin's potential as a safe and effective antibacterial agent.</p>","PeriodicalId":20506,"journal":{"name":"Probiotics and Antimicrobial Proteins","volume":" ","pages":""},"PeriodicalIF":4.4000,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"A Novel Type I Crustin Isoform from Scylla olivacea and its Antibacterial Potential: Membrane Depolarization, Disruption, and Induction of Reactive Oxygen Species as Modes of Action.\",\"authors\":\"S Neelima, M V Anju, K Archana, V V Anooja, P P Athira, M R Revathy, M Dhaneesha, A Muneer, T P Sajeevan, S Muhammed Musthafa, I S Bright Singh, S Muraleedharan Nair, Rosamma Philip\",\"doi\":\"10.1007/s12602-025-10469-7\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Availability of novel antimicrobial agents is an urgent necessity to combat the growing threat posed by multidrug-resistant bacteria, prompting exploration of marine antimicrobial peptides (AMPs) as potential solutions. Crustacean AMPs are equitably diverse in terms of structure and function, making them consistent templates for novel antimicrobials. From Scylla olivacea gill transcriptome cDNA, a putative Crustin AMP sequence of 333 nucleotides, encoding a 111 amino acid Crustin Type-I isoform was identified. The mature peptide encoding region was cloned and recombinantly expressed in E. coli using Luria Bertani (LB) broth, yielding approximately 0.9 mg/L peptide. This cationic (+6.25) peptide with amphipathic properties (34% hydrophobicity) exhibited antibacterial effects against Gram-positive and Gram-negative strains, with MIC 16 μM against Vibrio spp. The identified modes of actions included disruption of bacterial membranes, membrane potential dissipation, and induction of ROS. Scanning electron microscopy (SEM) analysis revealed bacterial lysis and structural damage. Being non-toxic to mammalian cells (CHO-K1) and non-haemolytic, So-Crustin qualifies to be safe for therapeutic applications. It was quite stable under different physio-chemical/biological conditions, including temperature, pH, NaCl concentrations and proteases like trypsin and proteinase K. This study emphasizes So-Crustin's potential as a safe and effective antibacterial agent.</p>\",\"PeriodicalId\":20506,\"journal\":{\"name\":\"Probiotics and Antimicrobial Proteins\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":4.4000,\"publicationDate\":\"2025-02-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Probiotics and Antimicrobial Proteins\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1007/s12602-025-10469-7\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Probiotics and Antimicrobial Proteins","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1007/s12602-025-10469-7","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
A Novel Type I Crustin Isoform from Scylla olivacea and its Antibacterial Potential: Membrane Depolarization, Disruption, and Induction of Reactive Oxygen Species as Modes of Action.
Availability of novel antimicrobial agents is an urgent necessity to combat the growing threat posed by multidrug-resistant bacteria, prompting exploration of marine antimicrobial peptides (AMPs) as potential solutions. Crustacean AMPs are equitably diverse in terms of structure and function, making them consistent templates for novel antimicrobials. From Scylla olivacea gill transcriptome cDNA, a putative Crustin AMP sequence of 333 nucleotides, encoding a 111 amino acid Crustin Type-I isoform was identified. The mature peptide encoding region was cloned and recombinantly expressed in E. coli using Luria Bertani (LB) broth, yielding approximately 0.9 mg/L peptide. This cationic (+6.25) peptide with amphipathic properties (34% hydrophobicity) exhibited antibacterial effects against Gram-positive and Gram-negative strains, with MIC 16 μM against Vibrio spp. The identified modes of actions included disruption of bacterial membranes, membrane potential dissipation, and induction of ROS. Scanning electron microscopy (SEM) analysis revealed bacterial lysis and structural damage. Being non-toxic to mammalian cells (CHO-K1) and non-haemolytic, So-Crustin qualifies to be safe for therapeutic applications. It was quite stable under different physio-chemical/biological conditions, including temperature, pH, NaCl concentrations and proteases like trypsin and proteinase K. This study emphasizes So-Crustin's potential as a safe and effective antibacterial agent.
期刊介绍:
Probiotics and Antimicrobial Proteins publishes reviews, original articles, letters and short notes and technical/methodological communications aimed at advancing fundamental knowledge and exploration of the applications of probiotics, natural antimicrobial proteins and their derivatives in biomedical, agricultural, veterinary, food, and cosmetic products. The Journal welcomes fundamental research articles and reports on applications of these microorganisms and substances, and encourages structural studies and studies that correlate the structure and functional properties of antimicrobial proteins.