Production of native flagellin from Salmonella Typhimurium in a bioreactor and purification by tangential ultrafiltration

Abstract

-Flagellin is the structural protein and most abundant component of bacterial flagella. The flagellum filament contains around 20,000 -100,000 subunits of 50 kDa flagellin that can have diverse biotechnological applications such as vaccine adjuvant and cellular protector during chemo-and radiotherapy. The main aim of this work was to study a production process of purified native FliC flagellin of Salmonella Typhimurium. The culture conditions in shakers were established with medium devoid of animal-derived components. In bioreactors, culture conditions were established in order to obtain flagellin from the culture supernatant by tangential ultrafiltration (TUF). The concentrated 750 kDa cut-off TUF fraction had a purification factor of 1.5 and a recovery yield of 52.2% for flagellin. The volumetric production of flagellin using the described procedure achieved around 307 mg/L of culture, which represented a significant improvement over previously reported methods. These results permit the development of production and purification processes that can be easily scaled up.

Escherichia coli‐based cell free production of flagellin and ordered flagellin display on virus‐like particles

“…Since bacterial flagellin has potential as a vaccine adjuvant, a rDNA flagellin production technology would be beneficial. Currently, flagellin proteins are produced by homogenization, heat treatment, and filtration of the flagellin filament pellets prepared from cell pellets (Braga et al, 2008; Oliveira et al, 2011; Zhang et al, 2003). Abdulla et al (2012) expressed the flagellin gene flic from S. typhimurium into E. coli .…”

Mapping Salmonella typhimurium pathways using 13C metabolic flux analysis

“…Besides the recognized importance as a pathogen itself, in the last years, Salmonella has gained increasing attention in the biotechnological area as a potential host to produce several pharmaceutical compounds (Silva et al, 2014). Among the products that can be produced with Salmonella typhimurium are flagellin, capsular polysaccharide Vi, lipopolysaccharides, and vaccines, with applications in human and veterinary medicine (Braga et al, 2010;Kong et al, 2013;Kothari et al, 2014;Oliveira et al, 2011). Attenuated strains of Salmonella are being used as Live Bacterial Vectors (LBV), for immunization against itself or to deliver heterologous antigens (Silva et al, 2014).…”

Chemical Characterization and Serological Activity of Ralstonia solanacearum Flagellin Preparatuons

“…Так, наибольшее число клеток R. solanacearum, имеющих жгутики, на- При химической идентификации препаратов флагеллинов показано (табл.1), что они состоят из белка (55-92%), углеводов (7-25%), а также в незначительных количествах обнаружены нуклеиновые кислоты (от 0.5 до 3.3%). Известно [16], что флагеллины построены из четырех структурных доменов: терминальные cпиральные D0 (включающий С-и N-концы) и D1, локализованные внутри нити, а также D2 и D3 домены, которые находятся на ее поверхности. Изучение первичной структуры флагеллинов из различных видов бактерий свидетельствует о высокой степени гомологии аминокислотных последовательностей их D0 и D1 доменов -80 и 60% гомологии соответственно и всего 20% гомологии в центральном домене.…”