Raman spectroscopic and laser scanning confocal microscopic analysis of sulfur in living sulfur-precipitating marine bacteria

Laser Raman microprobe spectroscopy and laser scanning confocal microscopy were used to determine the presence and speciation of sulfur in sulfur-oxidizing, marine bacteria from Monterey Bay, CA. The bacteria studied include: large, filamentous Thioploca and Beggiatoa, endosymbionts in the vesicomyid clam Calyptogena kilmeri, and a filamentous bacterium of undetermined species. All of these bacteria were shown spectroscopically to store elemental sulfur in submicrometer to several micrometer diameter vesicles. More detailed Raman spectroscopic study of the vesicles in Thioploca and Beggiatoa provided further chemical and structural characterization of the elemental sulfur. The sulfur is bonded in the common, stable S. ring configuration and is of an extremely fine-grained microcrystalline form. No additional (organo) sulfur compounds were detected spectroscopically in the vesicles under the low laser powers required to preserve the molecular structure of the sulfur. The present spectroscopic and optical data stand in contrast to reports and inferences of liquid-like elemental sulfur or homogeneous, complex sulfur compounds in other sulfur-oxidizing bacteria. The findings of this study are compatible with a model of sulfur vesicles as dominated by microcrystalline solid elemental sulfur, perhaps embedded in a matrix and/or confining membrane of organic material. The high reactivity and solubility observed in these vesicles is attributed to the extremely fine grain size of the solid elemental sulfur. (C) 2001 Elsevier Science B.V. All rights reserved.