RESUMO
This research investigated the use of single-walled carbon nanotubes (SWNTs) as an additive to increase the permeability of a bacterial cell wall. Recombinant Escherichia coli BL21 (DE3) that expressed ß-lactamase were exposed to SWNTs under various levels of concentration and agitation. Activity of ß-lactamase in the culture fluid and transmission electron microscopy (TEM) were used to determine the amount of released protein, and visually examine the permeability enhancement of the cells. It was found that ß-lactamase release in the culture fluid occurred in a dose-dependent manner with treatment by SWNTs and was also dependent on agitation rate. Based on TEM, this treatment successfully caused an increase in permeability without significant damage to the cell wall. Consequently, SWNTs can be used as an enhancement agent to cause the release of intracellular proteins. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:654-657, 2017.
Assuntos
Escherichia coli/metabolismo , Nanotecnologia/métodos , Nanotubos de Carbono/ultraestrutura , Microscopia Eletrônica de Transmissão , beta-Lactamases/metabolismoRESUMO
Interest in peptides as diagnostic and therapeutic materials require their manufacture via either a recombinant or synthetic route. This study examined the former, where a recombinant fusion consisting of an antifungal peptide was expressed and isolated from Escherichia coli. Fed batch fermentation with E. coli harboring an arabinose-inducible plasmid produced the 12 residue anti-Candida peptide fused to the N-terminal of Green Fluorescent Protein (GFPUV ). The purification of the fusion protein, using ion-exchange chromatography, was monitored by using the intrinsic fluorescence of GFPUV . The recombinant antifungal peptide was successfully released by cyanogen bromide-induced cleavage of the fusion protein. The recombinant peptide showed the expected antifungal activity. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:865-871, 2016.