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Improvement of Aspergillus oryzae for hyperproduction of endoglucanase: expression cloning of cmc-1 gene of Aspergillus aculeatus
Authors:Muhammad Hamid Rashid  Muhammad Rizwan Javed  Takashi Kawaguchi  Jun-ichi Sumitani  Motoo Arai
Institution:(1) Enzyme Engineering Group, Industrial Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), P.O. Box 577, Jhang Road, Faisalabad, Pakistan;(2) Graduate School of Agriculture and Biological Sciences, Osaka Prefecture University, Sakai Osaka, 599-8531, Japan;(3) Research Institute for Advanced Science and Technology, Osaka Prefecture University, Sakai Osaka, 599-8531, Japan
Abstract:FI-Carboxymethylcellulase (cmc1; family 12) is one of the endoglucanases of Aspergillus aculeatus and consists of single polypeptide chain of 221 amino acids. The cmc1 gene was expressed in Aspergillus oryzae niaD300 (niaD) under promoter 8142. The plasmid pCMG14 carrying the cmc1 gene at PstI site was used as a source of the gene (920 bp) and Aspergillus oryzae was successfully transformed by the plasmid pNAN-cmc1 (harboring cmc1 gene). The plasmid was integrated in Aspergillus oryzae niaD300 genome at niaD locus and the transformed fungus constitutively produced very high amounts of endoglucanases when grown on glucose, maltose, soluble starch and wheat bran.
Keywords:Aspergillus aculeatus                      Aspergillus oryzae            Endoglucanase  Expression cloning
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