Design, expression and characterization of lactiscin—A novel broad-spectrum peptidic bacteriocin
3
Issued Date
2023-09-01
Resource Type
ISSN
18788181
Scopus ID
2-s2.0-85167784362
Journal Title
Biocatalysis and Agricultural Biotechnology
Volume
52
Rights Holder(s)
SCOPUS
Bibliographic Citation
Biocatalysis and Agricultural Biotechnology Vol.52 (2023)
Suggested Citation
Rukying N., Ajingi Y.S., Nokyod S., Usman J.N., Ruengvisesh S., Rattanarojpong T., Pason P., Angsuthanasombat C., Jongruja N. Design, expression and characterization of lactiscin—A novel broad-spectrum peptidic bacteriocin. Biocatalysis and Agricultural Biotechnology Vol.52 (2023). doi:10.1016/j.bcab.2023.102811 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/88816
Title
Design, expression and characterization of lactiscin—A novel broad-spectrum peptidic bacteriocin
Other Contributor(s)
Abstract
Bacteria-derived antimicrobial peptides known as peptidic bacteriocins offer a promising alternative to traditional antibiotics in the face of the emergence of multidrug-resistant bacteria. Here, a nucleotide sequence of the gene encoding Lactococcus lactis-derived peptidic bacteriocin designated as lactiscin selectively identified from the GenBank® database was synthesized with an added 6⋅His sequence and cloned into Escherichia coli. Upon low-temperature expression at 16 °C, the His-tagged peptide could be produced in both soluble form and insoluble inclusions. Efficient purification of the soluble His-tagged peptide was achieved via immobilized-Ni2+ affinity chromatography (IMAC) and its estimated molecular mass of ∼13.4 kDa was determined by tricine-sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified peptide was highly active against both Gram-positive and Gram-negative bacteria as it exhibited a minimal inhibitory concentration of 0.45 mg/mL, 0.15 mg/mL, 0.35 mg/mL and 0.45 mg/mL against. Escherichia coli, Vibrio parachemolyticus, Staphylococcus aureus and Micrococcus luteus, respectively. In addition, the lactiscin peptide still retained antimicrobial activity over a pH range of 3.0–12.0 and heat stability of 100 °C for 30 min. A membrane integrity study revealed that this peptidic bacteriocin was able to induce E. coli membrane permeabilization in a concentration-dependent manner, albeit it showed a negligible toxic effect on erythrocytic cells. Gel retardation assay demonstrated that the lactiscin bacteriocin could suppress the migration of genomic DNA extracted from pathogenic bacteria, suggesting the presence of bacteriocin-responsive binding genomic. Our findings of lactiscin—a novel broad-spectrum bacteriocin would be a valuable additive for the application of food industry as a potential bio-preservative.