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Solubilization, partial purification and properties of lipoxygenase from apples

Solubilisierung, partielle Reinigung und Eigenschaften von Lipoxygenase aus Äpfeln

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Zusammenfassung

Die Wirksamkeit einer Anzahl lösender Agentien auf die Extraktion von Apfel-lipoxygenase zeigt deutlich ihre membrangebundene Natur. Es stellt sich heraus, daß Triton X-100 (1% v/v) und Natriumcholat (75 mM) am wirksamsten sind. Auch während der darauffolgenden Reinigung mußte ein geeignetes Detergens (Triton X-100) hinzugefügt werden, um Aggregations-Phänomene zu verhindern. Dieses zeigt den integralen Charaketer des Enzyms.

Partielle Reinigung eines lipoxygenasehaltigen Extraktes aus Äpfeln wurde durch Gelfiltration über Sephadex G-50 and Sephacryl S-300 sf erreicht. Ionenaustausch-und Affinitäts-Chromatographie waren nicht sehr erfolgreich.

Es wurden einige Eigenschaften der teilweise gereinigten Lipoxygenase untersucht. Das Enzym zeigte optimale Aktivität bei pH 6,9 and oxidierte vorwiegend freie Linolsäure. Das scheinbare Molekulargewicht (206000 Dalton) and der Stokes' Radius (51,5 Å) wiesen auf eine starke Bindung mit einer bedeutenden Menge von Triton X-100 hin.

Summary

The effectiveness of a number of solubilizing agents upon the extraction of apple lipoxygenase clearly demonstrated its membrane-bound nature. Triton X-100 (1% v/v) and sodium cholate (75 mM) proved most useful. Also, during subsequent purification procedures a suitable detergent (Triton X-100) had to be included to prevent aggregation phenomena, indicating the integral character of the enzyme.

Partial purification of a lipoxygenase extract from apples was achieved by gel filtration on Sephadex G-50 and Sephacryl S-300 sf. Ion-exchange chromatography and affinity chromatography proved to be of little success.

Some characteristics of the partially purified lipoxygenase were determined. The enzyme showed optimal activity at pH 6.9 and preferably peroxidized free linoleic acid. Apparent molecular weight (206,000 dalton) and Stokes' radius (51.5 Å) indicated a strong association with a substantial amount of Triton X-100.

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Authors and Affiliations

  1. Laboratory of Food Preservation, Catholic University of Leuven, de Croylaan 42, B-3030, Heverlee, Belgium

    Marcel Feys, René De Mot, Willy Naesens & Paul Tobback

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  1. Marcel Feys
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  2. René De Mot
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  3. Willy Naesens
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  4. Paul Tobback
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Feys, M., De Mot, R., Naesens, W. et al. Solubilization, partial purification and properties of lipoxygenase from apples. Z Lebensm Unters Forch 174, 360–365 (1982). https://doi.org/10.1007/BF01459953

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