Abstract
The fungus Humicola lutea 120-5 cultivated in casein-containing media, in the presence or absence of inorganic phosphate (Pi), excretes three different molecular forms of acid phosphatase (with Mr values of approximately 140, 70 and 35 kDa). The enzyme forms were isolated and purified 30–100-fold by a procedure involving two steps of ion-exchange chromatography and Sephadex G-200 gel chromatography. It was found that the fungus excretes only one of the phosphatases with the highest Mr (140 kDa) during growth on medium with inorganic nitrogen source (NaNO3). This form (designed AcPh I) was assumed to be a constitutive, since it showed resistance to high Pi-concentrations (10 mM) and its biosynthesis was not affected by the type of nitrogen source (casein or NaNO3). The other two forms (AcPh II-70 and AcPh III-35 kDa) were competitively inhibited by Pi (K i = 0.5 and 0.2 mM, respectively) and were induced by casein. The K m values of AcPh I and AcPh II were estimated as 1.3 mM, while AcPh III showed a higher affinity for p-nitrophenylphosphate (pNPP) with K m of 0.5 mM. The AcPh I–III fractions demonstrated a pH optimum in the range of 4.5–4.8 and an optimal temperature of 55 °C using pNPP as a substrate.
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Aleksieva, P., Grigorov, I., Djerova, A., Sheremetska, P. & Tchorbanov, B. 1984 Submerged cultivation of fungal mutant strain Humicola lutea 120-5 producing acid proteinases. Acta Biotechnologica 4, 179-188.
Aleksieva, P. & Mutafov, S. 1997 Continuous culture of Humicola lutea 120-5 for acid proteinase production. World Journal of Microbiology and Biotechnology 13, 353-357.
Aleksieva, P., Dulguerova, G. & Tchorbanov, B. 1998 Method for obtaining of acid proteinase and acid phosphatase. Bulgarian Patent No. 61453.
Aleksieva, P. & Micheva-Viteva, S. 2000 Regulation of extracellular acid phosphatase biosynthesis by phosphates in proteinase producing fungus Humicola lutea 120-5. Enzyme and Microbial Technology 27, 570-575.
Andersch, M.A. & Szezypinski, A.J. 1947 Use of p-nitrophenyl phosphate as the substrate in determination of serum acid phosphatase. American Journal of Clinical Pathology 17, 571-574.
Anson, M.L. 1938 The estimation of pepsin, trypsin, papain and cathepsin with hemoglobin. Journal of General Physiology 22, 79-89.
Arnold, W.N., Garrison, R.G., Mann, L.C. & Wallance, D.P. 1988 The acid phosphatases of Thermoascus crustaceus, a thermophilic fungus. Microbios 54, 101-112.
Galabova, D.N., Vasileva-Tonkova, E.S. & Balasheva, M.A. 1994 Comparison of the phosphate-repressible and-constitutive acid phosphatases of Yarrowia lipolytica. World Journal of Microbiology & Biotechnology 10, 483-484.
Grigorov, I., Aleksieva, P., Djerova, A., Sheremetska, P. & Tchorbanov, B. 1983 Selection of gamma-ray mutant from a strain Humicola lutea 72, producing acid proteases. European Journal of Applied Microbiology and Biotechnology 17, 355-357.
Haas, H., Redl, B., Leitner, E. & Stoffer, G. 1991 Penicillium chrysogenum extracellular acid phosphatase: purification and biochemical characterization. Biochimica et Biophysica Acta 1074, 392-397.
Kyriakidis, S.M., Sakellaris, G. & Sotiroudis, T.G. 1993 Is protein phosphorylation a control mechanism for the degradation of caseins by lactic acid bacteria? The detection of an extracellular acid phosphatase acitivity. Letters in Applied Microbiology 16, 295-298.
Nahas, E., Terenzi, H.F. & Rossi, A. 1982 Effect of carbon source and pH on the production and secretion of acid phosphatase (EC 3.1.3.2) and alkaline phosphatase (EC 3.1.3.1) in Neurospora crassa. Journal of General Microbiology 128, 2017-2021.
Nozawa, S.R., Macceheroni, W., Stabeli, R.G., Thedei, G. & Rossi, A. 1998 Purification and properties of Pi-repressible acid phosphatases from Aspergillus nidulans. Phytochemistry 49, 1517-1523.
Nyc, J.F., Kadner, R.J. & Crocken, B.J. 1965 A repressible alkaline phosphatase in Neurospora crassa. Journal of Biological Chemistry 241, 1468-1472.
Sviridenko, Ya. Ya., Sviridenko, G.M., Surkov, B.A. & Krayushkin, V.A. 1987 Effect of dephosphorylation on the biochemical properties of casein. Molochnaya promyshlennost 8, 2527 (Russ).
Tarafdar, J.C., Rao, A.V. & Bala, K. 1988 Production of phosphatases by fungi isolated from desert soils. Folia Microbiologica 33, 453-457.
Tchorbanov, B., Aleksieva, P., Dambadarja, N. & Dyulgerova, G. 1996 Extracellular acid phosphatase as a minor enzyme during the production of proteinases by Humicola lutea 120-5. Acta Biotechnologica 16, 65-71.
Yoshida, H. & Tamiya, N. 1971 Acid phosphatases from Fusarium moniliforme. Purification and enzymatic properties. Journal of Biochemistry 69, 525-534.
Yoshida, H., Oikawa, S., Ikeda, M. & Reese, E.T. 1989 A novel acid phosphatase excreted by Penicillium funiculosum that hydrolyzes both phosphodiesters and phosphomonoesters with aryl leaving groups. Journal of Biochemistry 105, 794-798.
Zyla, K. 1990 Acid phosphatases purified from industrial waste mycelium of Aspergillus niger used to produce citric acid. Acta Biotechnologica 10, 319-327.
Zyla, K. 1993 The role of acid phosphatase activity during enzymic dephosphorylation of phytates by Aspergillus niger phytase. World Journal of Microbiology and Biotechnology 9, 117-119.
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Micheva-Viteva, S., Tchorbanov, B., Aleksieva, P. et al. Acid phosphatases excreted by Humicola lutea 120-5 in casein-containing medium. World Journal of Microbiology and Biotechnology 16, 859–863 (2000). https://doi.org/10.1023/A:1008939221447
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DOI: https://doi.org/10.1023/A:1008939221447