Open Access

ARTICLE

Heterologous expression of bacteriocin E-760 in Chlamydomonas reinhardtii and functional analysis

by Quezada-Rivera JJ, RE Soria-Guerra, FS Pérez-Juárez, L Martínez-González, SE Valdés- Rodríguez, NL Vasco-Méndez, JF Morales-Domínguez

1 Departamento de Química, Universidad Autónoma de Aguascalientes, Universidad No. 940, Ciudad Universitaria, C.P. 20131; Aguascalientes, Ags. México.
2 Facultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Zona Universitaria Poniente, C.P. 78290; San Luis Potosí, S.L.P., México.
3 Departamento de Biotecnología y Bioquímica, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Cinvestav-Unidad Irapuato. Km. 9.6 Libramiento Norte Carr. Irapuato-León 36824 Irapuato, Gto., México.

* Corresponding Author: J F Morales-Domínguez, e-mail:

Phyton-International Journal of Experimental Botany 2019, 88(1), 25-35. https://doi.org/10.32604/phyton.2019.04549

Abstract

The use of antimicrobial peptides (AMPs) synthesized by bacteria (bacteriocins) is an alternative for combating multidrug resistant bacterial strains and their production by recombinant route is a viable option for their mass production. The bacteriocin E-760 isolated from the genus Enterococcus sp. has been shown to possess inhibitory activity against Gram-negative and Gram-positive bacteria. In this study, the expression of a chimeric protein coding for E-760 in the nucleus of C. reinhardtii was evaluated, as well as, its antibacterial activity. The synthetic gene E-760S was inserted into the genome of C. reinhardtii using Agrobacterium tumefaciens. A transgenic line was identified in TAP medium with hygromycin and also by PCR. The increment in the culture medium temperature of the transgenic strain at 35 °C for 10 minutes, increased the production level of the recombinant protein from 0.14 (Noninduced culture, NIC) to 0.36% (Induced culture, IC) of total soluble proteins (TSP); this was quantified by an ELISA assay. Recombinant E-760 possesses activity against Staphylococcus aureus in 0.34 U log, Streptococcus agalactiae in 0.48 U log, Enterococcus faecium in 0.36 U log, Pseudomonas aeruginosa in 2 U log and for Klebsiella pneumoniae, the activity was 0.07 U log. These results demonstrate that the nucleus transformation of C. reinhardtii can function as a stable expression platform for the production of the synthetic gene E-760 and it can potentially be used as an antibacterial agent.

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