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Enzymatic peptide synthesis in organic media : a comparative study of water-miscible and water-immiscible solvent systems

Clapés, Pere ; Adlercreutz, Patrick LU and Mattiasson, Bo LU (1990) In Journal of Biotechnology 15(4). p.323-338
Abstract

Peptide synthesis was carried out in a variety of organic solvents with low contents of water. The enzyme was deposited on the support material, celite, from an aqueous buffer solution. After evaporation of the water the biocatalyst was suspended in the reaction mixtures. The chymotrypsin-catalyzed reaction between Z-Phe-OMe and Leu-NH2 was used as a model reaction. Under the conditions used ([Z-Phe-OMe]0 ≤ 40 mM, [ Leu-NH2]0 ([Z-Phe-OMe]0 = 1.5) the reaction was first order with respect to Z-Phe-OMe. Tris buffer, pH 7.8, was the best buffer to use in the preparation of the biocatalyst. In water-miscible solvents the reaction rate increased with increasing water content, but the... (More)

Peptide synthesis was carried out in a variety of organic solvents with low contents of water. The enzyme was deposited on the support material, celite, from an aqueous buffer solution. After evaporation of the water the biocatalyst was suspended in the reaction mixtures. The chymotrypsin-catalyzed reaction between Z-Phe-OMe and Leu-NH2 was used as a model reaction. Under the conditions used ([Z-Phe-OMe]0 ≤ 40 mM, [ Leu-NH2]0 ([Z-Phe-OMe]0 = 1.5) the reaction was first order with respect to Z-Phe-OMe. Tris buffer, pH 7.8, was the best buffer to use in the preparation of the biocatalyst. In water-miscible solvents the reaction rate increased with increasing water content, but the final yield of peptide decreased due to the competing hydrolysis of Z-Phe-OMe. Among the water-miscible solvents, acetonitrile was the most suitable, giving 91% yield with 4% (by vol.) water. In water-immiscible solvents the reaction rate and the product distribution were little affected by water additions in the range between 0% and 2% (vol. %) in excess of water saturation. The reaction rates correlated well with the log P values of the solvent. The highest yield (93%) was obtained in ethyl acetate; in this solvent the reaction was also fast. Under most reaction conditions used the reaction product was stable; secondary hydrolysis of the peptide formed was normally negligible. The method presented is a combination of kinetically controlled peptide synthesis (giving high reaction rates) and thermodynamically controlled peptide synthesis (giving stable reaction products).

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author
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organization
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type
Contribution to journal
publication status
published
subject
keywords
Bioorganic synthesis, Enzymatic peptide synthesis, Organic media for enzymatic synthesis, α-Chymotrypsin
in
Journal of Biotechnology
volume
15
issue
4
pages
16 pages
publisher
Elsevier
external identifiers
  • scopus:0025483040
  • pmid:1366830
ISSN
0168-1656
DOI
10.1016/0168-1656(90)90090-X
language
English
LU publication?
yes
id
deb363b5-d16f-4802-acdd-b6cdd023274d
date added to LUP
2019-06-22 18:53:07
date last changed
2020-03-22 06:32:25
@article{deb363b5-d16f-4802-acdd-b6cdd023274d,
  abstract     = {<p>Peptide synthesis was carried out in a variety of organic solvents with low contents of water. The enzyme was deposited on the support material, celite, from an aqueous buffer solution. After evaporation of the water the biocatalyst was suspended in the reaction mixtures. The chymotrypsin-catalyzed reaction between Z-Phe-OMe and Leu-NH<sub>2</sub> was used as a model reaction. Under the conditions used ([Z-Phe-OMe]<sub>0</sub> ≤ 40 mM, [ Leu-NH<sub>2</sub>]<sub>0</sub> ([Z-Phe-OMe]<sub>0</sub> = 1.5) the reaction was first order with respect to Z-Phe-OMe. Tris buffer, pH 7.8, was the best buffer to use in the preparation of the biocatalyst. In water-miscible solvents the reaction rate increased with increasing water content, but the final yield of peptide decreased due to the competing hydrolysis of Z-Phe-OMe. Among the water-miscible solvents, acetonitrile was the most suitable, giving 91% yield with 4% (by vol.) water. In water-immiscible solvents the reaction rate and the product distribution were little affected by water additions in the range between 0% and 2% (vol. %) in excess of water saturation. The reaction rates correlated well with the log P values of the solvent. The highest yield (93%) was obtained in ethyl acetate; in this solvent the reaction was also fast. Under most reaction conditions used the reaction product was stable; secondary hydrolysis of the peptide formed was normally negligible. The method presented is a combination of kinetically controlled peptide synthesis (giving high reaction rates) and thermodynamically controlled peptide synthesis (giving stable reaction products).</p>},
  author       = {Clapés, Pere and Adlercreutz, Patrick and Mattiasson, Bo},
  issn         = {0168-1656},
  language     = {eng},
  month        = {01},
  number       = {4},
  pages        = {323--338},
  publisher    = {Elsevier},
  series       = {Journal of Biotechnology},
  title        = {Enzymatic peptide synthesis in organic media : a comparative study of water-miscible and water-immiscible solvent systems},
  url          = {http://dx.doi.org/10.1016/0168-1656(90)90090-X},
  doi          = {10.1016/0168-1656(90)90090-X},
  volume       = {15},
  year         = {1990},
}