Pepsin‐catalyzed peptide synthesis in organic media : studies with free and immobilized enzyme
BEMQUERER, MARCELO PORTO ; ADLERCREUTZ, PATRICK LU
and TOMINAGA, MINEKO
(1994)
In International Journal of Peptide and Protein Research
44(5).
p.448-456
- Abstract
Pepsin‐catalyzed synthesis of protected peptides was studied in two‐phase systems containing up to 50% (by volume) of aqueous phase. A methodological study was carried out to determine the optimum conditions for the synthesis of the model protected peptide Z‐Phe‐Phe‐OMe. Several parameters such as concentrations of carboxylic and amino components, pH of the aqueous phase, ratio of organic to aqueous phase volumes and nature of the organic solvent were investigated. It was observed that the most hydrophobic solvents produced the best yields, despite the low solubility of substrates in these media. The log P of the solvent could be used to predict the solvent effect over the reaction yields. Pepsin immobilized by adsorption onto the solid... (More)
Pepsin‐catalyzed synthesis of protected peptides was studied in two‐phase systems containing up to 50% (by volume) of aqueous phase. A methodological study was carried out to determine the optimum conditions for the synthesis of the model protected peptide Z‐Phe‐Phe‐OMe. Several parameters such as concentrations of carboxylic and amino components, pH of the aqueous phase, ratio of organic to aqueous phase volumes and nature of the organic solvent were investigated. It was observed that the most hydrophobic solvents produced the best yields, despite the low solubility of substrates in these media. The log P of the solvent could be used to predict the solvent effect over the reaction yields. Pepsin immobilized by adsorption onto the solid supports Celite and Chromosorb was employed to perform a study of secondary specificity of the enzyme in organic media through the coupling between Z‐X‐Phe‐OH (X = Ala, Asp, Glu, Gly, Phe, Ile, Val, Trp and Tyr) and Phe‐OMe. This investigation was performed in two solvent systems: (A) ethyl acetate:citrate buffer pH 4.5 (98:02, v:v) and (B) acetonitrile:citrate buffer pH 4.5 (96:04, v:v). Reaction rate data showed that pepsin had a preference for more hydrophilic substituents in the P2 position. These data are in contrast to the literature for a similar reaction performed in predominantly aqueous media. Thus, for mainly organic media, partition phenomena are very important and may cause an apparent modification of enzyme specificity.
(Less)
- author
- BEMQUERER, MARCELO PORTO
; ADLERCREUTZ, PATRICK
LU
and TOMINAGA, MINEKO
- organization
- publishing date
- 1994-01-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Celite, enzymatic, enzymes in organic media, free and immobilized pepsin, pepsin, peptide synthesis
- in
- International Journal of Peptide and Protein Research
- volume
- 44
- issue
- 5
- pages
- 9 pages
- publisher
- Munksgaard Forlag
- external identifiers
-
- pmid:7896503
- scopus:0027996504
- ISSN
- 0367-8377
- DOI
- 10.1111/j.1399-3011.1994.tb00181.x
- language
- English
- LU publication?
- yes
- id
- 87f8b2f0-a9b5-40f8-934c-3e8189958930
- date added to LUP
- 2019-06-22 09:15:08
- date last changed
- 2024-01-01 12:16:42
@article{87f8b2f0-a9b5-40f8-934c-3e8189958930,
abstract = {{<p>Pepsin‐catalyzed synthesis of protected peptides was studied in two‐phase systems containing up to 50% (by volume) of aqueous phase. A methodological study was carried out to determine the optimum conditions for the synthesis of the model protected peptide Z‐Phe‐Phe‐OMe. Several parameters such as concentrations of carboxylic and amino components, pH of the aqueous phase, ratio of organic to aqueous phase volumes and nature of the organic solvent were investigated. It was observed that the most hydrophobic solvents produced the best yields, despite the low solubility of substrates in these media. The log P of the solvent could be used to predict the solvent effect over the reaction yields. Pepsin immobilized by adsorption onto the solid supports Celite and Chromosorb was employed to perform a study of secondary specificity of the enzyme in organic media through the coupling between Z‐X‐Phe‐OH (X = Ala, Asp, Glu, Gly, Phe, Ile, Val, Trp and Tyr) and Phe‐OMe. This investigation was performed in two solvent systems: (A) ethyl acetate:citrate buffer pH 4.5 (98:02, v:v) and (B) acetonitrile:citrate buffer pH 4.5 (96:04, v:v). Reaction rate data showed that pepsin had a preference for more hydrophilic substituents in the P<sub>2</sub> position. These data are in contrast to the literature for a similar reaction performed in predominantly aqueous media. Thus, for mainly organic media, partition phenomena are very important and may cause an apparent modification of enzyme specificity.</p>}},
author = {{BEMQUERER, MARCELO PORTO and ADLERCREUTZ, PATRICK and TOMINAGA, MINEKO}},
issn = {{0367-8377}},
keywords = {{Celite; enzymatic; enzymes in organic media; free and immobilized pepsin; pepsin; peptide synthesis}},
language = {{eng}},
month = {{01}},
number = {{5}},
pages = {{448--456}},
publisher = {{Munksgaard Forlag}},
series = {{International Journal of Peptide and Protein Research}},
title = {{Pepsin‐catalyzed peptide synthesis in organic media : studies with free and immobilized enzyme}},
url = {{http://dx.doi.org/10.1111/j.1399-3011.1994.tb00181.x}},
doi = {{10.1111/j.1399-3011.1994.tb00181.x}},
volume = {{44}},
year = {{1994}},
}