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L-asparaginase of Thermus thermophilus: Purification, properties and identificaation of essential amino acids for its catalytic activity

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Abstract

L-asparaginase EC 3.5.1.1 was purified to homogeneity from Thermus thermophilus. The apparent molecular mass of L-asparaginase by SDS-PAGE was found to be 33 kDa, whereas by its mobility on Sephacryl S-300 superfine column was around 200 kDa, indicating that the enzyme at the native stage acts as hexamer. The purified enzyme showed a single band on acrylamide gel electrophoresis with pI = 6.0. The optimum pH was 9.2 and the Km for L-asparagine was 2.8 mM. It is a thermostable enzyme and it follows linear kinetics even at 77°C. Chemical modification experiments implied the existence of histidyl, arginyl and a carboxylic residues located at or near active site while serine and mainly cysteine seems to be necessary for active form.

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  1. Laboratory of Biochemistry, Department of Chemistry, Aristotle University of Thessaloniki, Thessaloniki, 54006, Greece

    Agathi A. Pritsa & Dimitrios A. Kyriakidis

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  1. Agathi A. Pritsa
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  2. Dimitrios A. Kyriakidis
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Correspondence to Dimitrios A. Kyriakidis.

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Pritsa, A.A., Kyriakidis, D.A. L-asparaginase of Thermus thermophilus: Purification, properties and identificaation of essential amino acids for its catalytic activity. Mol Cell Biochem 216, 93–101 (2001). https://doi.org/10.1023/A:1011066129771

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