ISOLATION OF (4R)-4-[(E)-2-BUTENYL]-4-METHYL-L-THREONINE, THE CHARACTERISTIC STRUCTURAL ELEMENT OF CYCLOSPORINS, FROM A BLOCKED MUTANT OF TOLYPOCLADIUM INFLATUM

Abstract

By mutagenic treatment of a strain of Tolypocladium inflatum, a cyclosporin non-producing mutant was obtained which accumulated the characteristic building unit of cyclosporins, (4R)-4-[(E)-2-butenyl]-4-methyl-L-threonine (abbreviation Bmt; systematic name: (2S, 3R, 4R, 6E)-2-amino-3-hydroxy-4-methyl-6-octenoic acid) in free form. The isolation from a culture filtrate was performed by extraction, chromatographic separation and final crystallization from methanol - water. The structure and stereochemistry of this amino acid was determined by chemical transformation and correlation to dihydro-MeBmt, with known chirality [(2S, 3R, 4R)-3-hydroxy-4-methyl-2-methylamino-octanoic acid], obtained by hydrolysis of dihydrocyclosporin A.