Characterization of an extracellular asparaginase of Rhodosporidium toruloides CBS14 exhibiting unique physicochemical properties

An asparaginase specific for L-asparagine was purified from Rhodosporidium toruloides CBS14 to apparent homogeneity. The enzyme was associated with L-glutaminase activity (Km, 1.43 × 10−3 M for L-asparagine and 6.45 × 10−3 M for L-glutamine). The enzyme was found to be a homodimer with a subunit molecular mass of 87 kDa. Chemical modification of tryptophan residues significantly reduced both L-asparaginase and L-glutaminase activities of the enzyme, which was prevented by the presence of either of the substrates L-asparagine or L-glutamine. The pH and temperature optima for both activities were 6.35 and 37 °C. The enzyme was serologically identical with the asparaginases of some of the other Rhodotorula and Rhodosporidium yeasts but was distinct from the asparaginase of Saccharomyces cerevisiae.Key words: asparaginase, glutaminase, Rhodosporidium toruloides.

[1]  M. Jaskólski,et al.  Crystal structure of Escherichia coli L-asparaginase, an enzyme used in cancer therapy. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[2]  C. Derst,et al.  Probing the role of threonine and serine residues of E. coli asparaginase II by site-specific mutagenesis. , 1992, Protein engineering.

[3]  S. Roy,et al.  Purification and properties of an L-asparaginase from Cylindrocarpon obtusisporum MB-10. , 1990, Biochemistry international.

[4]  J. Martín,et al.  Characterization and partial purification of L-asparaginase from Corynebacterium glutamicum. , 1990, Journal of general microbiology.

[5]  M P Gallagher,et al.  Asparaginase as a drug for treatment of acute lymphoblastic leukaemia. , 1989, Essays in biochemistry.

[6]  S. Jerebzoff-Quintin,et al.  Cyclic activity of L‐asparaginase through reversible phosphorylation in Leptosphaeria michotii , 1984, FEBS letters.

[7]  R. J. Roon,et al.  Derepression of asparaginase II during exponential growth of Saccharomyces cerevisiae on ammonium ion. , 1982, Archives of biochemistry and biophysics.

[8]  R. J. Roon,et al.  Reactions of asparaginase II of Saccharomyces cerevisiae. A mechanistic analysis of hydrolysis and hydroxylaminolysis. , 1980, The Journal of biological chemistry.

[9]  D. Ban,et al.  Characterization of two forms of asparaginase in Saccharomyces cerevisiae. , 1978, The Journal of biological chemistry.

[10]  B. Miflin,et al.  The purification and properties of asparaginase from Lupinus species , 1978 .

[11]  K. Arima,et al.  Partial Purification and Some Properties of Extracellular Asparaginase from Candida utilis , 1977 .

[12]  K. Arima,et al.  Extracellular Asparaginase from Candida utilis, Its Properties as Glycoprotein and Antitumor Activities , 1977 .

[13]  C. Drainas,et al.  Aspartic Hydroxamate Resistance and Asparaginase Regulation in the Fungus Aspergillus nidulans , 1977 .

[14]  R. Niederman,et al.  Purification and characterization of L-asparaginase with anti-lymphoma activity from Vibrio succinogenes. , 1976, The Journal of biological chemistry.

[15]  Y. Tamaura,et al.  Modification of cystine or tryptophan residues in the asparaginase molecule and their role in conformation and antigenicity , 1976, FEBS letters.

[16]  A. Goldstein,et al.  Purification and properties , 1975 .

[17]  J. Wriston,et al.  L-asparaginase: a review. , 2006, Advances in enzymology and related areas of molecular biology.

[18]  P. Joner,et al.  Purification and properties of L-asparaginase A from Acinetobacter calcoaceticus. , 1973, Biochimica et biophysica acta.

[19]  S. Igarasi,et al.  Asparaginase and glutaminase activities of micro-organisms. , 1973, Journal of general microbiology.

[20]  D. Miller,et al.  Physical properties and subunit structure of L-asparaginase isolated from Erwinia carotovora. , 1972, The Biochemical journal.

[21]  J. Wriston,et al.  Purification and properties of Bacillus coagulansl-asparaginase , 1971 .

[22]  B. Phillips,et al.  Asparaginase and glutaminase activities of bacteria. , 1971, Journal of general microbiology.

[23]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[24]  J. Wriston,et al.  Partial purification and antilymphoma activity of Serratia marcescensL-asparaginase☆ , 1967 .

[25]  A. Jeanes,et al.  Quantitative Determination of Monosaccharides as Their Alditol Acetates by Gas Liquid Chromatography. , 1965 .

[26]  J. Tepperman,et al.  ADAPTIVE HYPERLIPOGENESIS—LATE 1964 MODEL * , 1965, Annals of the New York Academy of Sciences.

[27]  W. J. Waddell,et al.  A simple ultraviolet spectrophotometric method for the determination of protein. , 1956, The Journal of laboratory and clinical medicine.

[28]  F. Smith,et al.  COLORIMETRIC METHOD FOR DETER-MINATION OF SUGAR AND RELATED SUBSTANCE , 1956 .