Heterologous expression of bacteriocin E-760 in Chlamydomonas reinhardtii and functional analysis
- Quezada-Rivera JJ1, RE Soria-Guerra2, FS Pérez-Juárez2, L Martínez-González2, SE Valdés- Rodríguez3, NL Vasco-Méndez1, JF Morales-Domínguez1
* Corresponding Author: J F Morales-Domínguez, e-mail:
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.
Keywords
Heterologous expression; Antibacterial activity; Bacteriocin E-760; Chlamydomonas reinhardtii; Log inactivation; Nuclear transformation.
Cite This Article
JJ, Q., Soria-Guerra, R., Pérez-Juárez, F., Martínez-González, L., Rodríguez, S. V. et al. (2019). Heterologous expression of bacteriocin E-760 in Chlamydomonas reinhardtii and functional analysis. Phyton-International Journal of Experimental Botany, 88(1), 25–35.
