Estudo da toxicidade do extrato da proteína PaOLP recombinante.

Abstract

Obesity is a global public health problem associated with several types of diseases and alterations, such as reductions in adiponectin levels. Research indicates that the osmotin protein is an agonist of the adiponectin hormone. This led to the isolation of a gene that encodes an osmotin-type protein, called PaOLP, which was cloned in the pQE30 vector to produce the recombinant mature protein, PaOLPHis6, in E. coli M15 cells. Aiming to complement the studies for potential biotechnological application, this study studied the toxicity of the soluble fraction of the PaOLPHis6 protein in Allium cepa roots. The concentration of total proteins present in the extracts from cells containing or not the gene for PaOLPHis6 (M15 PaOLP and M15 cells), with induced and non-induced expression, was determined by spectrophotometry. The roots were treated with extracts of induced and non-induced M15 PaOLP, induced and non-induced M15, in addition to the negative and positive control, for 72 hours. The toxicity analysis was performed using sections of the roots studied and the production of slides that were observed under an optical microscope. A total of 300 cells per slide were counted, and the number of cells in mitosis was recorded. Data analysis indicated that treatments with the extracts did not affect root growth or cell division. The presence of chromosomal and nuclear alterations was not observed, except in the positive control group. Therefore, the data from this study do not suggest toxicity to the extracts, and this represents relevant information. Future in vitro tests in animal cell cultures and in vivo in mice in vitro tests will contribute to the validation of the activity and toxicity of PaOLP.