Heptakis(2,6-O-dimethyl)beta-cyclodextrin: a novel growth stimulant for Bordetella pertussis phase I

Abstract

The effect of cyclodextrins on the growth of Bordetella pertussis Tohama phase I in synthetic medium was evaluated. The addition of cyclodextrins, especially heptakis(2,6-0-dimethyl)p-cyclodextrin (MePCD), to a complete synthetic medium such as Stainer-Scholte medium gave the same number of individual colonies and growth rates as those on Bordet-Gengou medium. Furthermore, with the addition of MePCD, growth inhibition by fatty acids such as oleic or palmitic acid was overcome and normal cell growth was observed. This modified Stainer-Scholte medium, designated as cyclodextrin solid medium (CSM), supported excellent growth of 20 lyophilized clinical isolates. Serotypes of the organisms after 10 passages on this CSM plate were not changed. These results suggest that MePCD is a significant growth stimulant and CSM is one of the most suitable synthetic media for culture of B. pertussis phase I.

Use of cyclodextrin as an agent to induce excretion of Bordetella pertussis antigens

“…Futhermore, bearing in mind that Me/3CD also enhances the release of the major component of outer membrane, namely the lipopolysaccharide (LPS), it is possible that cyclodextrin increases the level of extracellular antigens by stimulating their release, destabilising the outer membrane [12,24]. This hypothesis is in accordance with the observation that cyclodextrin forms inclusion complexes with hydrophobic compounds, such as fatty acids [8,9]. Since the lipopolysaccharide contains several fatty acids [25], Me/3CD may interact with this component of outer membrane.…”

“…The discovery that addition of 2,6-O-dimethyl-/3-cyclodextrin (Me/~CD) to the Stainer-Scholte (SS) [6] medium led to an enhanced level of AC, proved to be helpful [7]. Other authors have reported that cyclodextrin also increased the level of the other important soluble immunogens of B. pertussis such as pertussis toxin (PT) and filamentous hemagglutinin (FHA) [8][9][10][11]. However, the way which this compound enhances the yield of extracellular antigens, and in particular that of AC, remains to be elucidated.…”

Role ofBordetella pertussisRseA in the cell envelope stress response and adenylate cyclase toxin release

“…Liquid culture of B. pertussis used modified Stainer-Scholte (SS) medium (Stainer & Scholte, 1970;Schneider & Parker, 1982) supplemented with 0.5% casamino acids at 35°C with shaking at 300 r.p.m. SS cultures for b-galactosidase measurements also contained 500 lg mL À1 of heptakis (2,6-di-O-methyl)-b-cyclodextrin (Imaizumi et al, 1983). Escherichia coli strains were cultured on Luria Bertani agar or broth.…”

Secretion signal of the filamentous haemagglutinin, a model two‐partner secretion substrate

“…They were plated on Bordet Gengou (BG) agar (Bordet and Gengou, 1906) and incubated for 2 days at 37°C. Bacteria were used to inoculate small volumes of modified Stainer and Scholte (SS) medium (Imaizumi et al, 1983), and the cultures were grown overnight under orbital shaking at 37°C. These small cultures were used to inoculate larger volumes of modified SS medium, and the cultures were grown for 24 h under agitation at 37°C until late exponential phase.…”