Obtaining an Ent35-MccV derivative with mutated hinge region that exhibits increased activity against Listeria monocytogenes and Escherichia coli
The present paper describes the generation of derivatives from the hybrid peptide called Ent35-MccV, active against Gram-positive and Gram-negative bacteria. This peptide has a triple glycine hinge region between enterocin CRL35 and microcin V. In order to obtain variants of Ent35-MccV with greater biotechnological potential, a saturation mutagenesis was carried out in the hinge region. As a result, we obtained a bank of E. coli strains expressing different mutated hybrid bacteriocins in the central position of the hinge region. From all these variants, we found that the one bearing a tyrosine in the central region of the hinge (Ent35-GYG-MccV) is 2-fold more active against E. coli and 4-fold more active against Listeria than the original peptide Ent35-MccV. This derivative was purified and characterized. The development and evaluation of alternative hinges for Ent35-MccV represents a step forward in the bioengineering of antimicrobial peptides. This approach fosters the rational design of peptides with enhanced antimicrobial activity.
KeywordsENTEROCIN CRL35 MICROCIN V HYBRID PEPTIDE HINGE REGION MUTAGENESIS ANTIMICROBIAL ACTIVITY
This study was funded by grants PICT 4610 from the Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT), PIP 06906 CO from the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) and PIUNT D641/1 from the Secretaría de Ciencia, Arte e Innovación Tecnológica (SCAIT) from Universidad Nacional de Tucumán (UNT). S.A.N, L.L, N.S.R.C, M.F.U, are recipients of CONICET fellowship. S.A.N has received an ANPCyT fellowship. L.A, A.B and M.C.CH are career investigators of CONICET. B.S.P is CPA form CONICET.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
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