Development of enzymatic process to synthesize biologically active peptides with a extractive separation of produc

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The novel method to synthesize biologically active peptides with a continuous use of soluble enzyme and a simultaneous operation of an enzymatic reaction with a product separation was developed in this study, and applied to the protease-catalyzed synthesis of the precursor of aspartame (Z-APM and F-APM)A) Continuous synthesis of Z-APMpeptides are synthesized with enzyme at its optimal pH,though the enzymatic synthesis of peptides at low pH has a lot of advantages for an industrial-scaled production, such as, suppression of nonenzymatic decomposition of amino ester substrate and less requirement of NaOH for pH adjustment. We synthesize Z-APM at low pH in aqueous/organic biphasic system and obtained more than 95% of yield in this study. The mathe mathematical model to predict the continuous synthesis of Z-APM with extractive reaction was proposed and agreed well with the experimental results.B) Continuous synthesis of F-APMN-Benzyloxycarbonyl-L-aspartic acid (Z-L-Asp) and L-phenylalanine methyl ester (L-PheOMe) were, in general, used as the substrates for the enzymatic synthesis of aspartame precursor. N-Formyl group (F-group), which was cheaper and could be removed more easily than Z-group, was examined to be used for the protection of aspartic acid instead of Z-group to lower the total cost of the industrially continuous production of aspartame in this study. The F-APM yield was about 3% in aqueous monophsic system. The extremely high conversion, more than 95%, was, however, obtained for the thermolysin-catalyzed synthesis of F-APM in aqueous/TBP biphasic system.