Genetic engineering for obtaining cephalosporin production in Penicillium chrysogenum strain

ULLAN RV; TEJEIRA F; CASQUEIRO J.,; SCERVINO JM; MARTIN JF

Madrid, España

Congreso; 2nd FEMS Congress of European Microbiologists; 2006

Penicillium chrysogenum is an excellent host for â -lactam antibiotic production, because it overexpresses the first two genes pcbAB and pcbC common for penicillin and cephalosporin biosynthesis. We have introduced in P. chrysogenum the genetic information from Acremonium chrysogenum neccesary for the biosynthesis of cephalosporins. P. chrysogenum Wis54 npe6, a strain lacking the last step of penicillin biosynthesis has been transformed with the cefD1, cefD2, cefEF and cefG genes from A. chrysogenum. The recombinant strains TA64, TA71 and TA98 produce a higher amount of total âb- lactams than P. chrysogenum Wis 54-1255 in in complex production medium. The recombinant strains TA64, TA71 and TA98 produce extracellularly a high amount of deacetylcephalosporin C (DAC) but cephalosporin C (CPC) is not detected in the broths. Low DACacetyltransferase activity is present in TA71 and TA64 strains while TA98 showed a higher enzymatic activity. HPLC analysis revealed that TA64, TA71 and TA98 strains produced intracellularly a high concentration of DAC; moreover, TA64 and TA71 produce an small amount of intracellular CPC while HPLC analysis of TA98 revealed that it produces a moderate intracellular amount of CPC. Mass spectra analysis confirmed that TA98 produce true intracellular DAC and CPC.is an excellent host for â -lactam antibiotic production, because it overexpresses the first two genes pcbAB and pcbC common for penicillin and cephalosporin biosynthesis. We have introduced in P. chrysogenum the genetic information from Acremonium chrysogenum neccesary for the biosynthesis of cephalosporins. P. chrysogenum Wis54 npe6, a strain lacking the last step of penicillin biosynthesis has been transformed with the cefD1, cefD2, cefEF and cefG genes from A. chrysogenum. The recombinant strains TA64, TA71 and TA98 produce a higher amount of total âb- lactams than P. chrysogenum Wis 54-1255 in in complex production medium. The recombinant strains TA64, TA71 and TA98 produce extracellularly a high amount of deacetylcephalosporin C (DAC) but cephalosporin C (CPC) is not detected in the broths. Low DACacetyltransferase activity is present in TA71 and TA64 strains while TA98 showed a higher enzymatic activity. HPLC analysis revealed that TA64, TA71 and TA98 strains produced intracellularly a high concentration of DAC; moreover, TA64 and TA71 produce an small amount of intracellular CPC while HPLC analysis of TA98 revealed that it produces a moderate intracellular amount of CPC. Mass spectra analysis confirmed that TA98 produce true intracellular DAC and CPC.