Cloning and Characterization of Two Closely Linked Cellulase Genes from Cellvibrio mixtus

A 16.5-kb BamHI fragment of the Cellvibrio mixtus chromosome was found to direct carboxymethylcellulase, xylanase, and avicel hydrolysis. Two closely linked genes were subcloned from this insert. The gene, cmcI, was cloned as a 2.7-kb fragment and expressed in Escherichia coli. It encoded an enzyme of approximately 74 kDa which degraded carboxymethylcellulose and xylan but did not attack the microcrystalline cellulose substrate avicel. A second cellulase capable of degrading avicel, encoded by exoI, was found 5.5 kb downstream of cmcI. Two translation products of 53.7 kDa and 51.5 kDa were produced in E. coli strains expressing exoI. Northern analysis of total mRNA of C. mixtus grown on avicel, with a probe generated from cmcI, showed that cmcI and exoI were not cotranscribed in an operon.