Useful methods in enzymatic synthesis of peptides : A comparative study focussing on kinetically controlled synthesis of Ac-Phe-Ala-NH2 catalyzed by α-chymotrypsin
(1999) In Biocatalysis and Biotransformation 17(5). p.319-345- Abstract
The usefulness of different reaction systems for enzymatic peptide synthesis was evaluated by studying the formation of a dipeptide from equimolar amounts of acyl-donor and acyl-acceptor. α-Chymotrypsin (EC 3.4.21.1.), dissolved or deposited on celite,was used for the synthesis of Ac-Phe-Ala-NH2 from Ac-Phe-OEt and H-Ala-NH2. Reactions were carried out at room temperature (≃ 25°C), if not otherwise stated, in aqueous solutions (≤ 21% yield), frozen solutions at -30°C (≤ 60% yield), suspensions in aqueous media with and without 0.5% (v/v) of Triton X-100 (≤ 82% yield), suspensions in hydrophobic organic media with Na2CO3 . 10H2O as water source (≤ 90% yield), eutectic... (More)
The usefulness of different reaction systems for enzymatic peptide synthesis was evaluated by studying the formation of a dipeptide from equimolar amounts of acyl-donor and acyl-acceptor. α-Chymotrypsin (EC 3.4.21.1.), dissolved or deposited on celite,was used for the synthesis of Ac-Phe-Ala-NH2 from Ac-Phe-OEt and H-Ala-NH2. Reactions were carried out at room temperature (≃ 25°C), if not otherwise stated, in aqueous solutions (≤ 21% yield), frozen solutions at -30°C (≤ 60% yield), suspensions in aqueous media with and without 0.5% (v/v) of Triton X-100 (≤ 82% yield), suspensions in hydrophobic organic media with Na2CO3 . 10H2O as water source (≤ 90% yield), eutectic mixtures at 37°C (≤ 97% yield), and acetonitrile containing 1-5% (v/v) of aqueous buffer (≤ 99% yield). Initial rates of synthesis were in the range of 5.9 x 10-2-7.1 x 102 μmol min-1 (mg of enzyme)-1. The lowest one was obtained with dissolved reactants in acetonitrile containing 1% (v/v) of aqueous buffer, and the highest one was obtained with suspended acyl-donor in aqueous medium containing 0.5% (v/v) of Triton X-100.
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- author
- Björup, Peter ; Adlercreutz, Patrick LU and Clapés, Pere
- organization
- publishing date
- 1999-01-01
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Ac-Phe-Ala-NH, Eutectic mixtures, Frozen solutions, Organic media, Peptide synthesis, Surfactant, α-chymotrypsin
- in
- Biocatalysis and Biotransformation
- volume
- 17
- issue
- 5
- pages
- 27 pages
- publisher
- Taylor & Francis
- external identifiers
-
- scopus:0041467273
- ISSN
- 1024-2422
- DOI
- 10.3109/10242429909015234
- language
- English
- LU publication?
- yes
- id
- 84ca8af9-1544-4b7f-9f7c-81199d75fdb3
- date added to LUP
- 2019-06-20 15:18:43
- date last changed
- 2022-01-31 22:13:19
@article{84ca8af9-1544-4b7f-9f7c-81199d75fdb3, abstract = {{<p>The usefulness of different reaction systems for enzymatic peptide synthesis was evaluated by studying the formation of a dipeptide from equimolar amounts of acyl-donor and acyl-acceptor. α-Chymotrypsin (EC 3.4.21.1.), dissolved or deposited on celite,was used for the synthesis of Ac-Phe-Ala-NH<sub>2</sub> from Ac-Phe-OEt and H-Ala-NH<sub>2</sub>. Reactions were carried out at room temperature (≃ 25°C), if not otherwise stated, in aqueous solutions (≤ 21% yield), frozen solutions at -30°C (≤ 60% yield), suspensions in aqueous media with and without 0.5% (v/v) of Triton X-100 (≤ 82% yield), suspensions in hydrophobic organic media with Na<sub>2</sub>CO<sub>3</sub> <sup>.</sup> 10H<sub>2</sub>O as water source (≤ 90% yield), eutectic mixtures at 37°C (≤ 97% yield), and acetonitrile containing 1-5% (v/v) of aqueous buffer (≤ 99% yield). Initial rates of synthesis were in the range of 5.9 x 10<sup>-2</sup>-7.1 x 10<sup>2</sup> μmol min<sup>-1</sup> (mg of enzyme)<sup>-1</sup>. The lowest one was obtained with dissolved reactants in acetonitrile containing 1% (v/v) of aqueous buffer, and the highest one was obtained with suspended acyl-donor in aqueous medium containing 0.5% (v/v) of Triton X-100.</p>}}, author = {{Björup, Peter and Adlercreutz, Patrick and Clapés, Pere}}, issn = {{1024-2422}}, keywords = {{Ac-Phe-Ala-NH; Eutectic mixtures; Frozen solutions; Organic media; Peptide synthesis; Surfactant; α-chymotrypsin}}, language = {{eng}}, month = {{01}}, number = {{5}}, pages = {{319--345}}, publisher = {{Taylor & Francis}}, series = {{Biocatalysis and Biotransformation}}, title = {{Useful methods in enzymatic synthesis of peptides : A comparative study focussing on kinetically controlled synthesis of Ac-Phe-Ala-NH<sub>2</sub> catalyzed by α-chymotrypsin}}, url = {{http://dx.doi.org/10.3109/10242429909015234}}, doi = {{10.3109/10242429909015234}}, volume = {{17}}, year = {{1999}}, }