PROTEASE-CATALYZED SYNTHESIS OF OLIGOPEPTIDES IN HETEROGENOUS SUBSTRATE MIXTURES

A systematic study of enzymatic peptide synthesis in heterogenous subs trate mixtures was carried out, with the aim of establishing the prepa rative scope of this methodology. Semiliquid eutectics were obtained w ith various combinations of neutral, acidic, and basic amino acid deri vatives, in the presence or absence of adjuvants. A range of serine, c ysteine, and metalloproteases readily catalyzed the formation of the r equired dipeptides under these conditions. The synthetic usefulness of this approach was demonstrated by the sequential and convergent synth esis of derivatives of a number of bioactive di-, tri-, and pentapepti des, including aspartame, sweet lysine peptide, kyotorphin amide, ACE- inhibiting and -immunoactive tripeptides, and Leu-enkephalin amide, wi th overall yields of 21% to 84% and productivities of 0.13 to 0.75 g/g being obtained. (C) 1994 John Wiley & Sons, Inc.