- Email: [email protected]
5354667 Process for the enzymatic preparation of 7-Aminocephalosporanic acid
Fuels, Chemicals and Metals from Bioprocesses
5354667
5356794
PROCESS FOR THE ENZYMATIC PREPARATION OF 7AMINOCEPHALOSPORANIC ACID
PURIFIED ENZYMATIC PREPARATION DERIVED FROM CHROMOBACTERIUM VIOLACEUM, METHOD FOR PREPARING IT AND ITS USE IN THE PRODUCTION OF ALPHA, BETA° DEHYDROTRYPTOPHANYLPEPTIDES
Croux Clu-istian; Perez Javier C; Fuentes Jose L; Maldonado Francisco Madrid, SPAIN Assigned to Antibioticos S p A A process for the production of an E. coli strain producing high yields of 7 beta-(4carboxybutanamido) cephalosporin acylase, comprises: (a) digesting the DNA of a microorganism whine DNA includes the sequence encoding 7 beta- (4-earboxylmtanamido) cephalosporin acylase, and forming a plasmidic fibrary;(b) transforming, with the sequences in the plasmidic library,an auxotrephic E. coli host; (c) selecting for transformed E. coli hosts containing the acylase sequence by growth on a suitable medium; (d) isolating the vector containing the acylase sequence, digesting the vector, and ligating the DNA sequences obtained into an E. coil vector under control of an E. col/ promoter; (e) repeating selection procedure of step (c) ; (f) using the vectors from the selected E. coli hosts to transform an E. coli host lacking substantial betalactamase activity. 7-Aminocephalosporan/c acid and its derivatives can be prepared by reaction of substrates like 7 beta-(4-carboxybutanamido) cet~udosporan/c acid with 7 beta-(4catboxybetanamido) cephalosporin acylase produced by the E. coli strain produced by the above process.
Fromageot Pierre; Genet Roger le Chesnay, FRANCE Assigned to Commissariat a I~nergie Atomique Purified enzymatic preparation derived from Chromobacterium violaceum, having a dehydrogenase activity, method for preparing it and its use in the production of alpha, betadehydrou'yptophanyl-peptides. The said method for preparing alpha, beta-dehydrotryptophanylpeptides and alpha, beta-dehydrotryptophanylproteins is characterised in that one brings into contact: a peptide or a protein to be convened, and a purified enzymatic preparation having an Ltryptophan dehydroganase activity and having a specific activity of between 35 and 45 units.seconds-1 per mole of heme for N-acetyl-Ltryptophanamide (NATA), an apparent molecular weight of the order of 680 kDa, an isoelectric point of the order of 4, a temperature optimum for activity greater than 80 degrees C. and a temperature stability, at a pH of between 3 and 8 and at a temperature of between 20 degrees and 60 degrees C. Use of the dehydropeptides and dehydroproteins obtained as probes, reagents, drugs or cosmetic l~ducts.
825
5354667
5356794
PROCESS FOR THE ENZYMATIC PREPARATION OF 7AMINOCEPHALOSPORANIC ACID
PURIFIED ENZYMATIC PREPARATION DERIVED FROM CHROMOBACTERIUM VIOLACEUM, METHOD FOR PREPARING IT AND ITS USE IN THE PRODUCTION OF ALPHA, BETA° DEHYDROTRYPTOPHANYLPEPTIDES
Croux Clu-istian; Perez Javier C; Fuentes Jose L; Maldonado Francisco Madrid, SPAIN Assigned to Antibioticos S p A A process for the production of an E. coli strain producing high yields of 7 beta-(4carboxybutanamido) cephalosporin acylase, comprises: (a) digesting the DNA of a microorganism whine DNA includes the sequence encoding 7 beta- (4-earboxylmtanamido) cephalosporin acylase, and forming a plasmidic fibrary;(b) transforming, with the sequences in the plasmidic library,an auxotrephic E. coli host; (c) selecting for transformed E. coli hosts containing the acylase sequence by growth on a suitable medium; (d) isolating the vector containing the acylase sequence, digesting the vector, and ligating the DNA sequences obtained into an E. coil vector under control of an E. col/ promoter; (e) repeating selection procedure of step (c) ; (f) using the vectors from the selected E. coli hosts to transform an E. coli host lacking substantial betalactamase activity. 7-Aminocephalosporan/c acid and its derivatives can be prepared by reaction of substrates like 7 beta-(4-carboxybutanamido) cet~udosporan/c acid with 7 beta-(4catboxybetanamido) cephalosporin acylase produced by the E. coli strain produced by the above process.
Fromageot Pierre; Genet Roger le Chesnay, FRANCE Assigned to Commissariat a I~nergie Atomique Purified enzymatic preparation derived from Chromobacterium violaceum, having a dehydrogenase activity, method for preparing it and its use in the production of alpha, betadehydrou'yptophanyl-peptides. The said method for preparing alpha, beta-dehydrotryptophanylpeptides and alpha, beta-dehydrotryptophanylproteins is characterised in that one brings into contact: a peptide or a protein to be convened, and a purified enzymatic preparation having an Ltryptophan dehydroganase activity and having a specific activity of between 35 and 45 units.seconds-1 per mole of heme for N-acetyl-Ltryptophanamide (NATA), an apparent molecular weight of the order of 680 kDa, an isoelectric point of the order of 4, a temperature optimum for activity greater than 80 degrees C. and a temperature stability, at a pH of between 3 and 8 and at a temperature of between 20 degrees and 60 degrees C. Use of the dehydropeptides and dehydroproteins obtained as probes, reagents, drugs or cosmetic l~ducts.
825