Production of L-Asparaginase by Pseudomonas ovalis

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Summary

The enzyme L-asparaginase has been proved lately to have an inhibitory effect on human iymphosarcoma. Screening of microbial cultures, isolated from Egyptian soils, yielded a bacterium with high potentiality for L-asparaginase production. The organism was isolated on gelatin plates from samples, taken from slaughter house soils, and identified as Pseudomonas ovalis. The enzyme was assayed in intact cells by direct nesslerization of ammonia, liberated in the reaction. L-asparagine and L-aspartic and L-glutamic acids serve as inducers for the enzyme. The enzyme is produced optimum at pH 7.5 and repressed significantly by complex nitrogen sources and certain amino-acids. No activity could be detected in the presence of ammonium phosphate. The highest yield of the enzyme was obtained when the organism was grown aerobically in a medium containing L-glutamic acid and yeast extract, buffered at pH 7.5. Further characterization of the enzyme is reported.

Zusammenfassung

Es wurde festgestellt, daß das Enzym L-Asparaginase eine Hemmwirkung auf das menschliche Lymphosarkom ausüht. Ein Bakterium, das aus einem ägyptischen Boden isoliert wurde, stellt große Mengen an L-Aspariginase her. Der Organismus wurde aus Schlachthausboden über Gelatineplatten isoliert und als Pseudomonas ovalis identifiziert.

Die Enzymaktivität wurde in den Zellen durch direkte Neßlerisierung des gebildeten Ammoniaks ermittelt. L-Asparaginsäure, L-Asparagin und L-Glutaminsäure induzieren die Enzymbildung.

Das pH-Optimum für die Enzymbildung liegt hei 7,5. Manche N- Verbindungen, insbesondere Aminosäuren, hemmen die Enzymbildung.

In Gegenwart von Ammoniumphosphat wurde keine Aktivität nachgewiesen. Die höchste Ausbeute an Enzym wurde erzielt, wenn der Organismus aerob auf einem L-Glutaminsäure und Hefeextrakt enthaltenden Medium bei einem pH-Wert von 7,5 wachsen konnte.

Weitere Eigenschaften des Enzyms werden dargestellt.

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    Authors' address: Dr. Mohamed Salah Foda, Dr. Ekram Z. Khafagy, and Dr. Samir M. Badr El-Din, National Research Centre, Biochemistry and Microbial Chemistry Laboratories, Dokki, Giza (Egypt) (V.A.R.).

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