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Appl Environ Microbiol. 1992 June; 58(6): 1797-1800
Copyright © 1992, American Society for Microbiology. All Rights Reserved.

Preparation of Refractile Spores of Clostridium thermosaccharolyticum Involves a Solventogenic Phase

Sandra L. Landuyt and Edward J. Hsu^*

¹ Area of Microbiology, University of Missouri, Columbia, Missouri 65201, and School of Basic Life Sciences, University of Missouri, Kansas City, Missouri 64110²

ABSTRACT

Conversion of vegetative cells of Clostridium thermosaccharolyticum to refractile endospores was achieved by sequential transfer and dilution at each generation, with a final dilution into a sporulation medium that contained xylan supplemented with excess calcium. The subsequent growth was synchronous and resulted in elongated, solventogenic cells that were then shifted to 35°C to permit further differentiation without cell division. The synchronized cells grown in xylan medium supplemented with Ca gluconate produced total solvents that reached 9.63% (vol/vol). One hundred percent of these elongated solventogenic cells (4.84 x 10⁹ cells per ml) entered the sporangial stage and continued to differentiate into refractile spores. Only cells sequentially transferred and diluted at a critical time of the growth cycle are synchronized, induced to elongate (fourfold), become highly solventogenic in the presence of excess calcium, and are converted to a homogeneous population of refractile spores.

^* Corresponding author.

Present address: Department of Biology, Penn Valley Community College, Kansas City, MO 64111.