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Peptide synthesis by means of immobilized enzymes

Peptidsynthesen mit immobilisierten Enzymen. I. Immobilisiertes α-Chymotrypsin

I. Immobilized α-chymotrypsin

  • Organic Chemistry and Biochemistry
  • Published:
Monatshefte für Chemie / Chemical Monthly Aims and scope Submit manuscript

Abstract

α-Chymotrypsin covalently bound to silica, enzacryl AA, and enzacryl AH catalyzes peptide bond formation between N-protected dipeptide methyl esters and H-Leu-NH2 with results similar to those with the free enzyme. The influence of water-miscible and water-immiscible cosolvents, of the supports, and of the structure of the substrates is shown to be of importance for the ease of the chymotrypsin-medicated coupling reactions. The best yields were obtained using biphasic aqueous-organic solvent mixtures, silica-bound chymotrypsin, and substrates with leucine in the P2-position. The yields of the syntheses are discussed in terms of the reactivity of substrates with similar structure in enzymatic hydrolyses. All the immobilized chymotrypsin preparations could be re-utilized successfully for further couplings.

Zusammenfassung

Kovalent an Kieselgel, Enzacryl AA und Enzacryl AH gebundenes α-Chymotrypsin katalysiert die Peptidbindungsknüpfung zwischen N-geschützten Dipeptid-Methylestern und H-Leu-NH2 mit ähnlichen Ergebnissen wie das freie Enzym. Es wird gezeigt, daß mit Wasser mischbare und nichtmischbare Lösungsmittel, die Trägermaterialien sowie die Struktur der Substrate den Verlauf der Chymotrypsin-katalysierten Kupplungsreaktionen beeinflussen. Die besten Ausbeuten wurden in wäßrig-organischen Zweiphasen-systemen mit Kieselgel-gebundenem Chymotrypsin und mit Substraten mit Leucin in P2-Position erhalten. Die Syntheseausbeuten werden in bezug auf die Reaktivität von Substraten ähnlicher Struktur bei der enzymatischen Hydrolyse diskutiert. Alle immobilisierten Chymotrypsin-Präparationen konnten mit Erfolg für weitere Kupplungsreaktionen wiedergenutzt werden.

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Authors and Affiliations

  1. Sektion Biowissenschaften, Bereich Biochemie, Karl-Marx-Universität, DDR-701, Leipzig, German Democratic Republic

    Andreas Könnecke, Ralf Bullerjahn & Hans-Dieter Jakubke

Authors
  1. Andreas Könnecke
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  2. Ralf Bullerjahn
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  3. Hans-Dieter Jakubke
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Additional information

Abbreviations: IUPAC/IUB rules for peptides are followed, see Eur. J. Biochem.27, 201 (1972); Glt=4-carboxybutyrul (glutaryl),-Nan=4-nitroanilide. All amino acids except glycine are of L-configuration.

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Könnecke, A., Bullerjahn, R. & Jakubke, HD. Peptide synthesis by means of immobilized enzymes. Monatshefte für Chemie 112, 469–481 (1981). https://doi.org/10.1007/BF00901826

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  • DOI: https://doi.org/10.1007/BF00901826

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