Influence of culture medium on the production of eif antigen from Leishmania chagasi in recombinant Escherichia coli

Abstract

With the advent of recombinant DNA technology, recombinant protein expression has become an important tool in the study of the structure, function and identification of new proteins, especially those with therapeutic functions. Escherichia coli has been the predominant prokaryote used in genetic engineering studies due to the abundance of information about its metabolism. Despite significant advances in molecular biology and immunology of infections, there are as yet no prophylactic drugs capable of preventing visceral leishmaniasis. It is therefore important to identify specific antigens in order to develop vaccines and diagnostic kits against this disease. The objective of this study was to evaluate the influence of culture medium on the production of eIF antigen from Leishmania chagasi in recombinant Escherichia coli. An induction procedure using IPTG was carried out in a series of trials, to observe the influence of culture medium (2xTY, TB) under expression of the recombinant eIF protein. Results showed that recombinant protein expression was associated to growth and that the highest eIF antigen expression was obtained in the 2xTY medium.

Expression and production of recombinant scorpine as a potassium channel blocker protein in Escherichia coli

“…Phosphate buffer in TB media helps to keep pH constant during the time of bacterial growth . TB medium has a higher amount of yeast extract and peptone than LB, and increases the production of recombinant proteins . In TB + glucose culture medium, glucose was used instead of glycerol with the same carbon percent.…”

Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography

“…E. coli cultivation for 503 antigen production E. coli M15 strain expressing the His-tagged 503 antigen was kindly provided by Dr. Mary Wilson from the University of Iowa (Iowa, USA) [8]. The clone was cultured in 2xTY medium (16 g/L tryptone, 10 g/L yeast extract, and 5 g/L NaCl, pH 7.0) supplemented with ampicillin (0.1 g/L) and kanamycin (0.025 g/L) at 37 • C and 400 rpm on a bioreactor (Biostat B., B. Braun Biotech International) with a working volume of 1.5 L. The expression of the recombinant protein was induced by the addition of lactose to the cultivation medium at a final concentration of 10 g/L when the optical density at 600 nm reached approximately 0.5 [25]. Polypropylene glycol was used as an antifoam agent when necessary.…”

Production, recovery, and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography

“…The clone was cultured in 2xTY medium supplemented with antibiotics [5]. The cultivations were carried out using a bench bioreactor with a work volume of 1.5 L, at frequency of agitation of 400 rpm and constant output aeration of 1 vvm.…”