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Repeated batch and continuous synthesis of n benzyloxycarbonyl l phenylalanyl l phenylalanine methyl ester with immobilized thermolysin


Applied Microbiology & Biotechnology 28(3): 229-234
Repeated batch and continuous synthesis of n benzyloxycarbonyl l phenylalanyl l phenylalanine methyl ester with immobilized thermolysin
N-(Benzyloxycarbonyl)-L-phenylalanyl-L-phenylalanine methyl ester was synthesized from N-(benzyloxycarbonyl)-L-phenylalanine and L-phenylalanine methyl ester in an aqueous solution (aqueous phasic reaction), in an aqueous/organic biphasic system (biphasic reaction), and in an organic solvent (organic phasic reaction) with immobilized thermolysin. In the aqueous phasic reaction with thermolysin immobilized on Amberlite XAD-7, the whole product was trapped inside the support; extraction with ethyl acetate was needed to recover the product, and the equilibrium yield was low (about 65%). With the biphasic and organic phasic reactions with ethyl acetate as an organic solvent, the yield was around 95%. Because of the high yield and feasibility of operation, repeated batch and continuous reactions were done in the biphasic and organic phasic systems, respectively. The half-lives of the activity for the immobilized enzyme used in the biphasic system at 40.degree. C by repeated batch operation and in a plug flow reactor fed with substrate dissolved in ethyl acetate at 40.degree. C and 30.degree. C were estimated to be about 200 h (67 batches), 420 h, and 1100 h, respectively.

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Accession: 006312881



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