Unusually Stable Spinae from a Freshwater Chlorobium sp

A green Chlorobium sp. with spinae, strain JSB1, was isolated from an enrichment culture previously obtained from Fayetteville Green Lake, N.Y. (J. S. Brooke, J. B. Thompson, T. J. Beveridge, and S. F. Koval, Arch. Microbiol. 157:319-322, 1992). Cells were gram-negative, nonmotile rods which contained bacteriochlorophyll c and chlorosomes. Spinae were best seen by transmission electron microscopy in thin sections of cells fixed in the presence of tannic acid. High-resolution scanning electron microscopy showed the spinae randomly distributed at the cell surface and at the junctions between cells. Spinae were physically sheared from cells and isolated from the culture supernatant by ultrafiltration. As observed by electron microscopy, spinae demonstrated unusual structural stability when exposed for 1 h at 37 deg C to chemical treatments such as hydrogen bond-breaking agents, detergents, metal-chelating agents, proteases, and organic solvents. They were stable for 1 h at 37 deg C over the pH range 2.3 to 9.9 and in 1 M HCl and 1 M NaOH. The structural integrity of the spinae was also maintained when spinae were subjected to harsher treatments of autoclaving in 2% (wt/vol) sodium dodecyl sulfate and exposure to dithiothreitol at pH 9 for 1 h at 100 deg C. Partially dissociated spinae were obtained after 5 h at 100 deg C in 1 M HCl and 1 M NaOH. In acid, the tubular spinae became amorphous structures, with no helical striations visible. In alkali, the spinae had dissociated into irregular aggregates of disks. Since both high temperature and extremes of pH were required to achieve partial dissociation of the spinae, the strength of the structure presumably comes from covalent bonding.