Peptide synthesis with immobilized carboxypeptidase Y

Biotechnology and Bioengineering
S M Cramer, C Horváth

Abstract

Enzymatic peptide synthesis was investigated using carboxypeptidase Y immobilized with glutaraldehyde on 10 mum microparticulate amino-silica. Carboxypeptidase Y was immobilized with 98.5% recovery of active enzyme to yield the immobilized enzyme having 0.55 units esterase activity/mg amino-silica support. The stability of the immobilized enzyme was examined as a function of pH, temperature, and reactant concentrations. Immobilized Carboxypeptidase Y was used in stirred batch and recirculating packed-bed reactors for peptide synthesis. Packed-bed reactors (40 x 4.6 mm, 60 x 4.6 mm) were used to catalyze the synthesis of 170 mg N-benzoyl-L-arginyl-L-methioninamide, 380 mg N-benzoyl-L-arginyl-L-methionyl-L-leucinamide, and 200 mg N-benzoyl-L-arginyl-L-methionyl-L-leucyl-L-phenylalaninamide in 8, 3, and 1 hour, respectively, as intermediates in the synthesis of L-methionyl-L-leucyl-L-phenylalanine. No inactivation of the immobilized enzyme was observed during the course of the reactions. The N-benzoyl-L-arginyl group served to increase the water solubility of the peptides and was removed by immobilized trypsin at the end of synthesis to obtain the final product. While the first two syntheses were conducted with aqueous reaction mi...Continue Reading

References

Sep 1, 1977·Biotechnology and Bioengineering·A KlibanovI V Berezin
Apr 20, 1967·Biochemical and Biophysical Research Communications·I Schechter, A Berger
Jun 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·S I Wayne, J S Fruton
Sep 1, 1980·Journal of Biochemistry·T Oka, K Morihara
Sep 1, 1982·Applied Biochemistry and Biotechnology·I M ChaikenF Widmer

Citations

Aug 5, 1990·Biotechnology and Bioengineering·T W GusekJ E Kinsella
Jun 5, 1993·Biotechnology and Bioengineering·P S Sears, D S Clark
Jul 1, 1993·Biotechnology and Bioengineering·K KawashiroH Hayashi

Related Concepts

High Pressure Liquid Chromatography Procedure
Dimethylformamide
Glutaral
Immobilization
Peptides
Silicon Dioxide
Trypsin
E 2078
Methioninamide
Methionyl-leucyl-phenylalanine

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