Extracellular protease from (acremonium chrysogenum) with cpc-acetyl hydrolase activity and its utilization in the synthesis of deacetylated derivatives of cephalosporin c and inactivation of the gene for increasing production of cephalosporin

摘要

An extracellular protease of Acremonium chrysogenum with CPC-acetylhydrolase activity and its use for the synthesis of deacetylated derivatives of cephalosporin C and inactivation of the gene for increasing the yield of cephalosporin. The method includes the preparation of an enzyme with CPC-acetylhydrolase activity (CPC-AH B) starting from culture media of the filamentous fungus A. chrysogenum C10 (ATCC 48272) and the biochemical characterization of the said enzyme. In addition, it describes the cloning and sequencing of a genomic DNA fragment and another of complementary DNA (cDNA) which contain the cahB gene that codes for the said enzyme. By using the gene in the cDNA form, the production of CPC-AH B enzymatic activity in Escherichia coli is described. Finally, a method is included for increasing the yield of cephalosporin in A. chrysogenum by inactivation of the cahB gene.