Construction of Cellulase Producing Engineering Strains and the Application in Aged Tobacco Leaves
High cellulose content make the tissue of tobacco leaves broken easily,affect the plasticity of tobacco leaves during processing,and make the heavy impurity of tobacco leaves.In order to obtain excellent cellulase producing strains and achieve effective degradation of cellulose in aged tobacco leaves,ten Bacillus subtilis engineering strains of cellulose-degrading were suc-cessfully constructed by homologous recombination method.Four recombinant strains C36,CM,KF and GH5 with strong cellu-lase production capacity were screened by the Congo red plate method,carboxymethyl cellulose sodium enzyme activity assay,filter paper enzyme activity assay and filter paper disintegration rate detection.When aged tobacco leaves were used as substa-rate,the recombinant strain CM showed the highest cellulase production efficiency,and its carboxymethyl cellulose sodium en-zyme activity was 39.55 U·mL-1 and filter paper enzyme activity was 23.52 U·mL-1,respectively.The results indicated that the recombinant strains could utilize cellulose in aged tobacco leaves to produce cellulase,which could provide theoretical support for cellulose degradation of aged tobacco leaves in industrial production.
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