The prevalence of multidrug-resistant Gram-negative bacteria is a public health concern. Bacteriophages and bacteriophage-derived lytic enzymes have been studied in response to the emergence of multidrug-resistant bacteria. The availability of tRNAs and endolysin toxicity during recombinant protein expression is circumvented by codon optimization and lower expression levels using inducible pET-type plasmids and controlled cultivation conditions, respectively. The use of polyhistidine tags facilitates endolysin purification and alters antimicrobial activity. Outer membrane permeabilizers, such as organic acids, act synergistically with endolysins, but some endolysins permeate the outer membrane of Gram-negative bacteria per se. However, the outer membrane permeation mechanisms of endolysins remain unclear. Other strategies, such as the co-administration of endolysins with polymyxins, silver nanoparticles, and liposomes confer additional outer membrane permeation. Engineered endolysins comprising domains for outer membrane permeation is also a strategy used to overcome the current challenges on the control of multidrug-resistant Gram-negative bacteria. Metagenomics is a new strategy for screening endolysins with interesting antimicrobial properties from uncultured phage genomes. Here, we review the current state of the art on the heterologous expression of endolysin, showing the potential of bacteriophage endolysins in controlling bacterial infections.

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A B S TR A C TThe indiscriminate use of antibiotics and the emergence of resistant microorganisms have become a major challenge for the food industry. The purpose of this work was to microencapsulate the bacteriophage UFV-AREG1 in a calcium alginate matrix using microfluidic devices and to study the viability and efficiency of retention. The microcapsules were added to gel of propylene glycol for use as an antimicrobial in the food industry. The technique showed the number of the phage encapsulation, yielding drops with an average 100-250 µm of diameter, 82.1 ± 2% retention efficiency and stability in the gel matrix for 21 days. The gel added to the microencapsulated phage showed efficiency (not detectable on the surface) in reducing bacterial contamination on the surface at a similar level to antimicrobial chemicals (alcohol 70%). Therefore, it was possible to microencapsulate bacteriophages in alginate-Ca and apply the microcapsules in gels for use as sanitizers in the food industry.

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