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An anaerobic, mesophilic, sporeforming, xylanolytic bacterium was isolated from decayed Pinus patula wood chips. The cells of this organism were gram-negative rods, had peritrichous flagella, and formed terminal round endospores which distended the cell wall. Xylan, cellobiose, fructose, glucose, maltose, mannose, melezitose, raffinose, rhamnose, salicin, sucrose, and xylose were utilized. The optimum temperature and optimum pH for growth were 35°C and 7.2, respectively. The DNA composition was 40 mol% guanine plus cytosine. The name Clostridium xylanolyticum sp. nov. is proposed for this bacterium. The type strain is strain ATCC 49623. The potential use of biological hydrolysis of the hemicel- lulose components of wood has attracted much interest in the forestry and pulp and paper industries (9). After paper manufacture the hemicellulose fraction is essentially a waste product, but if it could be utilized efficiently by microorgan- isms, this fraction could become an important substrate for the production of several products, such as alcohol. Many xylanolytic organisms, mainly fungi and actinomycetes (1, 14-16), have been isolated from various environments. How- ever, no economically feasible system for biopulping has been developed (9). As a result of biopulping research in our laboratories, an obligately anaerobic bacterium was isolated from decayed Pinus patula wood chips. In this paper this isolate is proposed as a new species, Clostridium xylanolyti- cum. MATERIALS AND METHODS Media. The basal medium (BM) used in this study con- tained 1.2% (wt/vol) Bacto-Agar (Difco), 230 ml of clarified rumen fluid, 12.5 ml of mineral solution I (2), 12.5 ml of mineral solution I1 (2), 0.25 g of yeast extract (Merck), 1.0 g of peptone (Merck), 2.5 ml of a vitamin solution (3), 5.0 ml of a volatile fatty acid mixture (4), 5.0 ml of hemin (0.01%, wt/vol), 0.5 ml of Pfennig trace element solution (12), and 230 ml of distilled water. Sodium bicarbonate was used as a buffering agent, indigocarmine (0.05%, wt/vol) was used as a redox indicator, and 1.0 ml of cysteine monohydrochloride (12.5%, wt/vol) and 1.0 ml of Na,S (12.5%, wt/vol) were used as redox poisoning agents. The pH was adjusted to pH 6.8 with 1 N HC1 or 1 N NaOH as required. BM supple- mented with 3% oat spelt xylan (Fluka Chemicals) was also used, and for culture maintenance BM was supplemented with 0.3% cellobiose (Merck). Anaerobic culture methods. In this study an anaerobic cabinet (Forma Scientific, Marietta, Ohio) was used to culture organisms under strict anaerobic conditions. Cul- tures were maintained in 3-oz (ca. 90-ml) McCartney bottles that had been purged with an 0,-free gas mixture containing 5.5% H,, 31.8% CO,, and 62.7% N, prior to use and sealed with butyl rubber stoppers and screw caps. All cultures were incubated at 35°C without agitation. Isolation procedure. Samples of decayed P. patula wood

Clostridium xylanolyticum sp. nov., an Anaerobic Xylanolytic Bacterium from Decayed Pinus patula Wood Chips