Comparison of free and immobilized L-asparaginase synthesized by gamma-irradiated Penicillium cyclopium

• Autorzy: El-Refai H.A. , Shafei M.S. , Mostafa H. , El-Refai A.-M.H. , Araby E.M. , El-Beih F.M. , Easa S.M. , Gomaa S.K.
• Języki publikacji: EN

Abstrakty

EN

Gamma irradiation is used on Penicillium cyclopium in order to obtain mutant cells of high L-asparaginase productivity. Using gamma irradiation dose of 4 KGy, P. cyclopium cells yielded L-asparaginase with extracellular enzyme activity of 210.8 ± 3 U/ml, and specific activity of 752.5 ± 1.5 U/mg protein, which are 1.75 and 1.53 times, respectively, the activity of the wild strain. The enzyme was partially purified by 40–60% acetone precipitation. L-asparaginase was immobilized onto Amberlite IR-120 by ionic binding. Both free and immobilized enzymes exhibited maximum activity at pH 8 and 40°C. The immobilization process improved the enzyme thermal stability significantly. The immobilized enzyme remained 100% active at temperatures up to 60°C, while the free asparaginase was less tolerant to high temperatures. The immobilized enzyme was more stable at pH 9.0 for 50 min, retaining 70% of its relative activity. The maximum reaction rate (Vmax) and Michaelis-Menten constant (Km) of the free form were significantly changed after immobilization. The Km value for immobilized L-asparaginase was about 1.3 times higher than that of free enzyme. The ions K⁺, Ba²⁺ and Na⁺ showed stimulatory effect on enzyme activity with percentages of 110%, 109% and 106% respectively.

• Wydawca: -
• Czasopismo: Polish Journal of Microbiology
• Rocznik: 2016
• Tom: 65
• Numer: 1
• Opis fizyczny: p.45-50,fig.,ref.

Twórcy

autor

El-Refai H.A.

• Chemistry of Natural and Microbial Products Department, National Research Center, Dokki, Cairo, Egypt

autor

Shafei M.S.

• Chemistry of Natural and Microbial Products Department, National Research Center, Dokki, Cairo, Egypt

autor

Mostafa H.

• Chemistry of Natural and Microbial Products Department, National Research Center, Dokki, Cairo, Egypt

autor

El-Refai A.-M.H.

• Chemistry of Natural and Microbial Products Department, National Research Center, Dokki, Cairo, Egypt

autor

Araby E.M.

• Microbiology Department, Egyptian Atomic Energy Authority, Cairo, Egypt

autor

El-Beih F.M.

• Microbiology Department, Faculty of Science, Ein-Shams University, Cairo, Egypt

autor

Easa S.M.

• Microbiology Department, Faculty of Science, Ein-Shams University, Cairo, Egypt

autor

Gomaa S.K.

• Chemistry of Natural and Microbial Products Department, National Research Center, Dokki, Cairo, Egypt

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Identyfikatory

DOI

10.5604/17331331.1197274