EXTRACELLULAR PROTEASE FROM $I(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

The invention relates to an extracellular protease from Acremonium chrysogenum with CPC-acetyl hydrolase activity and its utilization in the synthesis of deacetylated derivatives of cephalosporin C and the inactivation of the gene for increasing the production of cephalosporin. Said method includes the preparation of an enzyme with CPC acetyl hydrolase (CPC-AH) activity from culture mediums of filamentous fungus A. chrysogenum C10 (ATCC 48272) and biochemical characterization of said enzyme. The invention also relates to the cloning and sequencing of a genomic DNA fragment and a complementary DNA (cDNA) fragment containing gene cahB coding for said enzyme. By using the gene in the form of a cDNA, the production of enzymatic activity CPC-AH B in Escherichia coli can be described. The invention further relates to a method for increasing the production of cephalosporin in A. chrysogenum by inactivating the gene cahB.