title: Eurystatins A and B, new prolyl endopeptidase inhibitors. III. Fermentation and controlled biosynthesis of eurystatin analogs by Streptomyces eurythermus.
abstract: Accurate and precise component analysis of eurystatin analogs in fermentation broth was devised by HPLC methods with and without 2,4-dinitrophenylhydrazonation. Detailed optimization of fermentation conditions and strain improvement by HPLC analysis significantly increased the eurystatin productivity of Streptomyces eurythermus. Chemically defined fermentation media which produced eurystatins A and B at fermentation yields comparable to complex media were elaborated for radio-isotope fermentation studies and controlled biosynthesis. Radio-isotope incorporation study using 14C-labeled amino acids in chemically defined medium demonstrated that L-leucine and L-ornithine were the direct precursors for the L-leucine and L-ornithine moieties of eurystatins A and B, respectively. Based on this finding, L-valine and L-isoleucine were supplemented to the growing culture of S. eurythermus in chemically defined medium, which resulted in the controlled biosynthesis of new eurystatin analogs named eurystatins C, D, E and F.
authors.affiliation: Bristol-Myers Squibb Research Institute, Tokyo, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Suzuki
authors.forename: S
authors.initials: S
authors.lastname: Toda
authors.forename: T
authors.initials: T
authors.lastname: Furumai
authors.forename: Y
authors.initials: Y
authors.lastname: Fukagawa
authors.forename: T
authors.initials: T
authors.lastname: Oki
mesh_terms: D000596:Amino Acids / Q000378:metabolism / Q000494:pharmacology; D002250:Carbon Radioisotopes; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D005285:Fermentation; D007532:Isoleucine / Q000378:metabolism; D015394:Molecular Structure; D010456:Peptides, Cyclic / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry; D010659:Phenylhydrazines; D000085822:Prolyl Oligopeptidases; D012697:Serine Endopeptidases / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors; D013302:Streptomyces / Q000378:metabolism*; D014633:Valine / Q000378:metabolism
chemical_list: D000596:Amino Acids; D002250:Carbon Radioisotopes; D003470:Culture Media; D010456:Peptides, Cyclic; D010659:Phenylhydrazines; D015842:Serine Proteinase Inhibitors; D007532:Isoleucine; C078927:eurystatin A; C078928:eurystatin B; C004787:2,4-dinitrophenylhydrazine; D012697:Serine Endopeptidases; D000085822:Prolyl Oligopeptidases; D014633:Valine