Enzymic peptide synthesis in microaqueous, solvent-free systems.
Thermolysin-catalyzed (EC 3.4.24.4) and chymotrypsin-catalyzed (EC 3.4.21.1) peptide synthesis reactions were accomplished without any organic solvent in the presence of low amounts of water under sonication and fluidization. The systems used are considered to be microaqueous solvent-free ones. The influence of several reaction parameters, such as time, the amount of enzyme, the amount of water in free form or bound as hydration water, and the N/C component ratio, on the vield of the thermolysin-catalyzed synthesis of Z-Phe-Leu-NH(2) (up to 87% yield) was investigated in a sonicated system. Besides Z-Phe-Leu-NH(2), the tripeptide derivatives Ac-Xaa-Trp-Leu-NH(2), (Xaa = Gly, Ala) were also obtained in good yields of 79 and 71% respectively. In the latter case, no hydrolytic side reactions were observed. Using a fluidized-bed reactor, chymotrypsin- and thermolysin-catalyzed syntheses of N-protected di- and tripeptide amides could be perfromed with yields in the range of 10 to 40%. (c) 1995 John Wiley & Sons, Inc.
Kuhl
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Technical University of Dresden, Institute of Biochemistry, Mommsenstrasse 13, D-01062 Dresden, Germany.
Elchhorn
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Jakubke
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