Continuous enzymatic production of oligopeptides: synthesis of an enkephalin pentapeptide in a multistage bioreactor.
A feasibility study of the continuous enzymatic production of short oligopeptides was undertaken using the synthesis of [Leu5]-enkephalin pentapeptides as a model system. A three-stage bioreactor was designed to perform the independent syntheses of the constituent tri- and dipeptide fragments and their subsequent condensation. Both the N-terminal (N-X-L-Tyr.Gly.GlyOEt) and C-terminal (L-Phe.LeuNH2) peptides were prepared in 75-85% yield in column reactors packed with Celite-immobilized alpha-chymotrypsin. The enkephalin pentapeptide was obtained in up to 30% yield in the third bioreactor module containing Celite-immobilized proteinase K. The bioreactor was run continuously for over 1,000 h, producing, under steady-state conditions, up to 0.7 g day-1 of the pentapeptide.
Richards
A O
AO
Department of Biotechnology and Enzymology, AFRC Institute of Food Research, Reading, UK.
Gill
I S
IS
Vulfson
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EN
D000595:Amino Acid Sequence; D004745:Enkephalins / Q000138:chemical synthesis*; D004798:Enzymes; D005240:Feasibility Studies; D007700:Kinetics; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis*
D004745:Enkephalins; D004798:Enzymes; D009842:Oligopeptides