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Screening, identification of an alcohol-tolerance cellulase-producing strain and its characteristics

An ethanol-tolerant strain Lys-1249 with cellulase activity was screened from the mature fermented grain mash by ethanol-CMC-Na plate preliminary screening and shake flask fermentation affination. Strain Lys-1249 was identified as Bacillus subtilis according to morphological observation, physiological and biochemical determination and 16 SrDNA sequence analysis. The cellulose-producing condition was optimized by orthogonal experiment, and results showed that the ethanol-tolerant cellulase activity reached 158.54 U/ml when strain Lys-1249 was cultured in the condition of wheat bran 2.0%, peptone 0.6%, K2HPO40.1%, Mg SO4·7H2O 0.05%, Fe SO47H2 O 0.02%, Mn SO40.01%, initial p H 7.0, temperature 37 ℃ and time48 h. The strain also grew well in medium with 6.0%vol ethanol, and the cellulase activity can keep at 69.95%.