Understanding protease catalysed solid phase peptide synthesis

R.V. Ulijn, N. Bisek, P.J. Halling, S.L. Flitsch

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

A protease (thermolysin) was used to directly synthesise a number of dipeptides from soluble Fmoc-amino acids onto a solid support (PEGA1900) in bulk aqueous media, often in very good yields. This shift in equilibrium toward synthesis is remarkable because for soluble dipeptides in aqueous solution hydrolysis rather than synthesis is observed. Three possible reasons for the equilibrium shift were considered: (i) using a solid support makes it easy to use an excess of reagents, so mass action contributes towards synthesis; (ii) reduction in the unfavourable hydrophobic hydration of the Fmoc group within the solid support compared with the free amino acid in solution and (iii) suppression of the ionization of amino groups linked to the solid phase due to mutual electrostatic repulsion. It was found that under the conditions studied the second effect was most important.
LanguageEnglish
Pages1277-1281
Number of pages4
JournalOrganic and Biomolecular Chemistry
Volume1
Issue number8
DOIs
Publication statusPublished - 2003

Fingerprint

protease
Solid-Phase Synthesis Techniques
Dipeptides
peptides
solid phases
Peptide Hydrolases
Thermolysin
Amino Acids
Peptides
amino acids
synthesis
Static Electricity
Hydrolysis
shift
reagents
hydration
hydrolysis
retarding
Hydration
electrostatics

Keywords

  • protease
  • dipeptides
  • Fmoc-amino acids
  • reagents
  • hydrophobic hydration
  • mutual electrostatic repulsion

Cite this

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abstract = "A protease (thermolysin) was used to directly synthesise a number of dipeptides from soluble Fmoc-amino acids onto a solid support (PEGA1900) in bulk aqueous media, often in very good yields. This shift in equilibrium toward synthesis is remarkable because for soluble dipeptides in aqueous solution hydrolysis rather than synthesis is observed. Three possible reasons for the equilibrium shift were considered: (i) using a solid support makes it easy to use an excess of reagents, so mass action contributes towards synthesis; (ii) reduction in the unfavourable hydrophobic hydration of the Fmoc group within the solid support compared with the free amino acid in solution and (iii) suppression of the ionization of amino groups linked to the solid phase due to mutual electrostatic repulsion. It was found that under the conditions studied the second effect was most important.",
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Understanding protease catalysed solid phase peptide synthesis. / Ulijn, R.V.; Bisek, N.; Halling, P.J.; Flitsch, S.L.

In: Organic and Biomolecular Chemistry, Vol. 1, No. 8, 2003, p. 1277-1281.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Understanding protease catalysed solid phase peptide synthesis

AU - Ulijn, R.V.

AU - Bisek, N.

AU - Halling, P.J.

AU - Flitsch, S.L.

PY - 2003

Y1 - 2003

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KW - protease

KW - dipeptides

KW - Fmoc-amino acids

KW - reagents

KW - hydrophobic hydration

KW - mutual electrostatic repulsion

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