2024-03-29T02:29:04+01:00 https://library.wur.nl/oai
oai:library.wur.nl:wurpubs/406626 2024-03-11
urn:nbn:nl:ui:32-406626 2024-03-11 urn:nbn:nl:ui:32-406626/mods Synthesis of Peptide Amides using Sol-Gel Immobilized Alcalase in Batch and Continuous Reaction System Corici L.N. aut Frissen A.E. 078252679 0000000396804187 aut van Zoelen D.J. aut Eggen I.F. aut Peter F. aut Davidescu C. aut Boeriu C.G. 314600051 0000-0003-4903-6621 0000000393659092 aut text info:eu-repo/semantics/article 2011 79955715090 en Two commercial proteases from Bacillus licheniformis (Alcalase 2.4 L FG and Alcalase 2.5 L, Type DX) were screened for the production of Z-Ala-Phe-NH2 in batch reaction. Alcalase 2.4 L FG was the most efficient enzyme for the C-terminal amidation of Z-Ala-Phe-OMe using ammonium carbamate as ammonium source. Immobilization of protease has been achieved by the sol-gel method, using dimethyldimethoxysilane (DMDMOS) and tetramethoxysilane (TMOS) as precursors (unpublished results). In batch production, about 95% of Z-Ala-Phe-NH2 was obtained at 30°C after 24 hours of incubation. Reproducibility of different batches of commercial Alcalase 2.4 L FG preparations was also investigated by evaluating the amidation activity and the entrapment yields in the case of immobilization. A packed-bed reactor (0.68 cm ID, 15.0 cm long) was operated successfully for the continuous synthesis of peptide amides. The immobilized enzyme retained the initial activity over 10 cycles of repeated use in continuous reactor at ambient temperature. At 0.75 mL/min flow rate of the substrate mixture, the total conversion of Z-Ala-Phe-OMe was achieved after 5 hours of substrate recycling. The product contained about 90% peptide amide and 10% hydrolysis byproduct. BBP Sustainable Chemistry & Technology Packed-bed reactor Peptide amide Protease Sol-gel immobilization BBP Sustainable Chemistry & Technology World Academy of Science, Engineering and Technology 5 4 794 799 20703740 urn:nbn:nl:ui:32-406626/obj 2024-03-11 http://purl.org/eprint/accessRights/RestrictedAccess