The phylogenetic tree of the partial elongation factor-1 alpha gene fits better than the partial 18S rDNA for generic classification. From the results of the molecular tree and analysis of morphological characters, Petriella setifera LH was identified. It can be induced to produce carboxymethyl cellulase (CMCase). The crude CMCase only shows a 44.1-kDa band by activity staining after SDS-PAGE. It is optimally active at 55°C and pH 6.0, and is stable from pH 5.0–8.0 and at 45°C or below. The crude CMCase, which is not affected by Co2+, is strongly activated in the presence of 10 mM Na+, K+, Ca2+, Mg2+, EDTA, and Mn2+. It is strongly inhibited by 10 mM Fe2+, Pb2+, Al3+, Zn2+, Ag+, Fe3+, and Cu2+. When compared with denim treatment by Novoprime A800 (a commercial neutral cellulase), crude CMCase exhibits a similar fabric weight loss and indigo dye removal. These results indicate that crude CMCase has potential application in denim biostoning.