CHARACTERIZATION OF ENGF, A GENE FOR A NON-CELLULOSOMAL CLOSTRIDIUM-CELLULOVORANS ENDOGLUCANASE

A new Clostridium cellulovorans (strain ATCC 35296) endoglucanase gene engF has been isolated and sequenced. The gene contains 1671 bp and c odes for a protein containing 557 amino acids and a mass of 60.1 kDa. A putative signal peptide of 29 amino acids is present and the mature protein has a mass of 57.1 kDa. EngF does not have amino acid sequence homology to previously isolated EngB and EngD, but does show sequence homology to family 5 glycosyl hydrolases from Bacillus, Erwinia carot ovora, and C. acetobulylicum species. EngF is not a component of the c ellulosome and does not contain a duplicated sequence (DS) at its C-te rminal region. EngF is capable of binding to cellulose and hydrolyzing carboxymethylcellulose but not xylan. The cellulose binding domain (C BD) differs from types I, II and III CBDs and no obvious homology has been found to other CBD types, The maximum activity of EngF occurs at pH 5.5 and at 47 degrees C. Its properties suggest that EngF plays an ancillary role in the degradation of cellulosic materials.