Clostridium saccharolyticum sp. nov., a Saccharolytic Species from Sewage Sludge

Abstract

A new species of Clostvidium isolated from a methanogenic cellulose-enrichment culture of sewage sludge is described. The colonies produced by these bacteria were white, circular, and convex with smooth margins. The cells were straight, spindle-shaped rods, 0.6 by 3.0 pm in size. They were gram negative and nonmotile, and they formed round, terminal spores. A wide variety of carbohydrates was fermented by this mesophilic anaerobe. The major fermentation products were acetic acid, hydrogen, carbon dioxide, and ethanol. The deoxyribonucleic acid base composition was 28 mol% guanine plus cytosine. The name Clostridiurn saccharolyticurn is proposed for this new species on the basis of its broad saccharolytic activity. The type strain of C. sczc.c.hLrrol\/ticirrn is WM1 ( = . . NRC 2533).A new species belonging to the genus Clostridiurn was isolated during the course of a study of the digestion of cellulose by a highly cellulolytic mixed culture. This culture was obtained from a methanogenic cellulose-enrichment culture started from sewage sludge (4, 5). In the present paper we define the characteristics of and propose a name for this newly discovered microorganism. MATERIALS AND METHODS Media.The basal medium used for the isolation and maintenance of the new isolate was the synthetic saltvitamin medium previously described by Khan et al. (6). The basal medium was supplemented with 0.1% yeast extract and was prepared by the procedure of Holdeman and Moore (1). The pH of the medium was adjusted to 7.2 to 7.4 prior to being reduced by the Hungate technique (2). For the preparation of media containing insoluble substrates, such as cellulose, the reduced medium was dispensed, under 80% N?-20% C 0 2 , in 10-ml volumes into 60-ml serum vials containing preweighed amounts of the substrate and was then autoclaved. Soluble substrates were filter sterilized and were injected by hypodermic syringe into the basal medium after the vials had been autoclaved and cooled. The isolated microorganism was maintained in basal medium supplemented with 2% (wthol) cellobiose and 0.1% yeast extract (YE).Isolation. A methanogenic cellulose-enrichment culture described by Khan (4) was used as the source of inoculum for isolation purposes. The inoculum was heated at 90°C for 15 min and was then serially diluted in basal medium. Prereduced cellobiose-agar slants, inside 160-ml serum vials, supplemented with 0.1% (wthol) YE, were spread with a 0.1-ml quantity of i. Issued as NRCC no. 19895. each dilution. The inoculated agar slants were flushed with an 80% N2-20% C 0 2 gas mixture, sealed, and incubated at 35°C. After 48 h, an individual colony was picked and transferred to cellobiose-YE broth. The broth was heated at 90°C for 15 min, serially diluted, and replated on cellobiose-YE agar. This procedure was repeated a total of five times.Biochemical tests. Biochemical tests were carried out in basal medium containing 0.1% YE and 2% of the selected substrate, by the procedures of Holdeman and Moore (1). The inoculated test vials were incubated at ...

Fusicatenibacter saccharivorans gen. nov., sp. nov., isolated from human faeces

“…Also, C. saccharolyticum WM 1 T (Murray et al, 1982) did not produce acid either from D-sorbitol or inulin and had no enzyme activity of esterase lipase (C8) and a-galactosidase. In addition, the DNA G+C content of strains HT03-11 T , .2 mol%) were far higher than that of C. saccharolyticum WM 1 T (28.0 mol%).…”

“…The type, and currently only, species of the genus is Fusicatenibacter saccharivorans. (Clavel et al, 2007;Song et al, 2003;Murray et al, 1982). +, Positive; 2, negative.…”

The Effect of Membrane Type on the Performance of Microbial Electrosynthesis Cells for Methane Production

“…44 Amongst the fermentative bacteria, phylotypes closer to Clostridium saccharolyticum/xylanolyticum were highly abundant in Nafion-BES compared to Ultrex-BES, in which their abundance amounted to less than 1% although many Clostridium phylotypes at lower than 1% relative abundance were present in the Ultrex-BES. These results may indicate that Clostridium saccharolyticum and/or other Clostridium spp.…”

Broad Distribution and Phylogeny of Anaerobic Endophytes of Cluster XIVa Clostridia in Plant Species Including Crops

“…Group I includes C. xylanolyticum from wood chips 32) , C. aerotolerans from rumens 42) , Clostridium algidixylanolyticum from raw lamb meat 4) , and C. saccharolyticum from a methanogenic cellulose-enrichment culture of sewage sludge 29) . The first three strains are all able to degrade xylan, whereas C. saccharolyticum uses cellobiose.…”