Abstract
Thermolysin–catalyzed condensation of N – (benzylxycarbyl) – glycyl – glycyl – L –phenylalanyl–L–leucine ethyl ester (Z–Gly–GlyPheLeuOEt), the precursor of the des–Tyr1–leucine enkephalin from N–(benzyl–oxycarbonyl)–glycyl–glycine (Z–GlyGly) and L–phenylalanyl–L–leucine ethyl ester (PheLeuOEt) was elucidated as a model reaction for the enzymatic synthesis of oligopeptides with many side reactions. In buffer solution, the yield of Z–GlyGly–PheLeuOEt was extremely low with considerable by–products. A large increase in both yield and purity became possible in the aqueous–organic biphasic system by the systematic selection of experimental conditions.
Similar content being viewed by others
References
Fruton, J.S. 1982. Proteinase-catalyzed synthesis of peptide bonds, p. 239–306. In: Adv. Enzymol., Vol. 53. A. Meister (ed.), John Wiley & Sons, New York.
Klibanov, A.M., Samokhin, G.P., Martinek, K., and Berezin, I.V. 1977. A new approach to preparative enzymatic synthesis. Biotechnol. Bioeng. 19:1351–1361.
Martinek, K. and Semenov, A. 1981. Enzymatic synthesis in biphasic aqueous-organic systems II. Shift of ionic equilibria. Biochim. Biophys. Acta 658:90–101.
Homandberg, G.A., Mattis, J.A., and Laskowski, M. Jr. 1978. Synthesis of peptide bonds by proteinases. Addition of organic co-solvents shifts peptide bond equilibria toward synthesis. Biochemistry 17:5220–5227.
Lüthi, P. and Luisi, P.L. 1984. Enzymatic synthesis of hydrocarbonsoluble peptides with reverse micelles. J. Am. Chem. Soc. 106:7285–7286.
Nakanishi, K., Kamikubo, T., and Matsuno, R. 1985. Continuous synthesis of N-(benzyloxycarbonyl)-L-aspartyl-L-phenylalanine methyl ester with immobilized thermolysin in an organic solvent. Bio/Technology 3:459–464.
Endo, S. 1962. Studies on protease produced by thermophilic bacteria. J. Ferment. Technol. 40:346–353.
Malfroy, B., Swerts, J.P., Guyon, A., Roques, B.P., and Schwartz, J.C. 1978. High-affinity enkephalin-degrading peptidase in brain is increased after morphine. Nature 276:523–526.
Zaks, A. and Klibanov, A.M. 1984. Enzymatic catalysis in organic media at 100°C. Science 224:1249–1251.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Nakanishi, K., Kimura, Y. & Matsuno, R. Design of Proteinase–Catalyzed Synthesis of Oligopeptides in an Aqueous–Organic Biphasic System. Nat Biotechnol 4, 452–454 (1986). https://doi.org/10.1038/nbt0586-452
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nbt0586-452
- Springer Nature America, Inc.
This article is cited by
-
An effective biphase system accelerates hesperidinase-catalyzed conversion of rutin to isoquercitrin
Scientific Reports (2015)