Skip to main content
SpringerLink
Log in

Purification and characterization of an acid phosphatase from Trichoderma harzianum

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

An acid phosphatase from Trichoderma harzianum was purified in a single step using a phenyl-Sepharose chromatography column. A typical procedure showed 22-fold purification with 56% yield. The purified enzyme showed as a single band on SDS-PAGE with an apparent molecular weight of 57.8 kDa. The pH optimum was 4.8 and maximum activity was obtained at 55°C. The enzyme retained 60% of its activity after incubation at 55°C for 60 min. The K m and V max values for p-nitrophenyl phosphate (p-NPP) as a substrate were 165 nM and 237 nM min−1, respectively. The enzyme was partially inhibited by inorganic phosphate and strongly inhibited by tungstate. Broad substrate specificity was observed with significant activities for p-NPP, ATP, ADP, AMP, fructose 6-phosphate, glucose 1-phosphate and phenyl phosphate.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Aleksieva P, Spasova D, Radoevska S (2003) Acid phosphatase distribution and localization in the fungus Humicola lutes. Zeitschrift fur Naturforschung C-A. J Biosci 58:239–243

    Google Scholar 

  • Almeida FB, Cerqueira FM, Silva RN, Ulhoa CJ, Lima AL (2007) Mycoparasitism studies of Trichoderma harzianum strains against Rhizoctonia solani: evaluation of coiling and hydrolytic enzyme production. Biotechnol Lett 29:1189–1193

    Article  CAS  PubMed  Google Scholar 

  • Altomare C, Norvell WA, Bjorkman T, Harman GE (1999) Solubilization of phosphates and micronutrients by the plant growth promoting and biocontrol fungus Trichoderma harzianum rifai 1295-22. Appl Environ Microbiol 65:2926–2933

    CAS  PubMed  Google Scholar 

  • Ames BN (1966) Assay of inorganic phosphate and phosphatases. Methods Enzymol 8:115–118

    Article  CAS  Google Scholar 

  • Bernard M, Mouyna I, Dubreucq G, Debeaupuis J-P, Fontaine T, Vorgias C, Fuglsang C, Latgé J-P (2002) Characterization of a cell-wall acid phosphatase (PhoAp) in Aspergillus fumigatus. Microbiology 148:2819–2829

    CAS  PubMed  Google Scholar 

  • Blum H, Beier H, Gross H (1987) Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8:93–99

    Article  CAS  Google Scholar 

  • Boyce A, Walsh G (2007) Purification and characterization of an acid phosphatase with phytase activity from Mucor hiemalis. J Biotechnol 132(1):82–87

    Article  CAS  PubMed  Google Scholar 

  • Dickman MB, Yarden O (1999) Serine/threonine protein kinases and phosphatases in filamentous fungi. Fungal Gen Biol 26:99–117

    Article  CAS  Google Scholar 

  • Duff SMG, Sarath G, Plaxton WC (1994) The role of acid phosphatases in plant phosphorus metabolism. Physiol Plant 90:791–800

    Article  CAS  Google Scholar 

  • Eherlich CK, Montalbano GB, Mullaney JE, Dischinger CH, Ullah AJH (1994) An acid phosphatase from Aspergillus ficuum has homology to Penicillium chrysogenum PHOA. Biochem Biophys Res Commun 204(1):63–68

    Article  Google Scholar 

  • Fujita J, Yamane Y, Fukuda H, Kizaki Y, Wakabayashi S, Shigeta S, Suzuki O, Ono K (2003) Production and properties of phytase and acid phosphatase from sake koji mold, Aspergillus oryzae. J Biosci Bioeng 95(4):348–353

    CAS  PubMed  Google Scholar 

  • Gargova S, Sariyska M (2003) Effect of culture conditions on the biosynthesis of Aspergillus niger phytase and acid phosphatase. Enzym Microb Technol 32:231–235

    Article  CAS  Google Scholar 

  • Guimarães LHS, Terenzi HF, Jorge JÁ, Leone JÁ, Polizeli MDTM (2004) Characterization and properties of acid phosphatases with phytase activity produced by Aspergillus caespitosus. Biotechnol Appl Biochem 40:201–207

    Article  PubMed  Google Scholar 

  • Haas H, Redl B, Leitner E, Stoffler G (1991) Penicillium chrysogenum extracellular acid phosphatase: purification and biochemical characterization. Biochim Biophys 1074(3):392–397

    CAS  Google Scholar 

  • Haran S, Logendra S, Seskar M, Bratanova M, Raskin I (2004) Characterization of Arabidopsis acid phosphatase promoter and regulation of acid phosphatase expression. Plant Physiol 124(2):615–626

    Article  Google Scholar 

  • Harman GE, Howell CR, Viterbo A, Chet I, Lorito M (2004) Trichoderma species—opportunistic, avirulent plant symbionts. Nat Rev Microbiol 2(1):43–56

    Article  CAS  PubMed  Google Scholar 

  • Jing G, Li L, Li Y, Xie L, Zhang R (2006) Purification and partial characterization of two acid phosphatase forms from pearl oyster (Pinctada fucata). Comp Biochem Physiol B 143(2):229–235

    Article  PubMed  Google Scholar 

  • Li Z, Wang Z, Peng G, Yin Y, Zhao H, Cao Y, Xia Y (2006) Purification and characterization of a novel thermostable extracellular protein tyrosine phosphatase from Metarhizium anisopliae strain CQMa102. Biosci Biotechnol Biochem 70(8):1961–1968

    Article  CAS  PubMed  Google Scholar 

  • Nozawa SR, MacCheroni W Jr, Stábeli RG, Thedei G Jr, Rossi A (1998) Purification and properties of pi-repressible acid phosphatases from Aspergillus nidulans. Phytochemistry 49(6):1517–1523

    Article  CAS  PubMed  Google Scholar 

  • Palacios MC, Haros M, Rosell CM, Sanz Y (2005) Characterization of an acid phosphatase from Lactobacillus pentosus: regulation and biochemical properties. J Appl Microbiol 98:229–237

    Article  CAS  PubMed  Google Scholar 

  • Rudresh DL, Shivaprakash MK, Prasad RD (2005) Tricalcium phosphate solubilizing abilities of Trichoderma spp. In relation to p uptake and growth and yield parameters of chickpea (Cicer arietinum L.). Can J Microbiol 51(3):217–222

    Article  CAS  PubMed  Google Scholar 

  • Tomaz CT, Queiroz JA (1999) Studies on the chromatographic fractionation of Thichoderma reesei cellulases by hydrophobic interaction. J Chromatogr A 865(1–2):123–128

    Article  CAS  PubMed  Google Scholar 

  • Wannet WJ, Wassenaar RW, Jorissen HJ, van der Drift C, Op den Drift C, Op den Camp HJ (2000) Purification and characterization of an acid phosphatase from the commercial mushroom Agaricus bisporus. Antonie Van Leeuwenhoek 77(3):215–220

    Article  CAS  PubMed  Google Scholar 

  • Wyss M, Pasamontes L, Rémy R, Kohler J, Kusznir E, Gadient M, Muller F, van Loon APGM (1998) Comparison of the thermostability properties of three acid phosphatases from molds: Aspergillus fumigatus phytase, Aspergillus niger phytase, and Aspergillus niger pH 2.5 acid phosphatase. Appl Environ Microbiol 64(11):4446–4451

    CAS  PubMed  Google Scholar 

  • Yadav SR, Tarafdar CJ (2003) Phytase and phosphatase producing fungi in arid and semi-arid soils and their efficiency in hydrolyzing different organic P compounds. Soil Biol Biochem 35:745–751

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by a biotechnology research grant to C. J. U. (CNPq/FINEP-MCT, CAPES/PROCAD, FAPEG/GO and FUNAPE/UFG). V. O. L. was supported by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior do Brasil.

Author information

Authors and Affiliations

  1. Universidade Federal de Goiás, Campus Samambaia, ICB II, Cx. Postal 131, Goiânia, GO, 74001-970, Brazil

    Vanessa Oliveira Leitão & Cirano J. Ulhoa

  2. Faculdade Padrão Av. Anhanguera esq. c/Rua do Algodão, Goiânia, GO, 74000-000, Brazil

    Rhalcia Cristina de Melo Lima

  3. Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, 91501-970, Brazil

    Marilene Henning Vainstein

Authors
  1. Vanessa Oliveira Leitão
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rhalcia Cristina de Melo Lima
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Marilene Henning Vainstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Cirano J. Ulhoa
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Cirano J. Ulhoa.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Leitão, V.O., de Melo Lima, R.C., Vainstein, M.H. et al. Purification and characterization of an acid phosphatase from Trichoderma harzianum . Biotechnol Lett 32, 1083–1088 (2010). https://doi.org/10.1007/s10529-010-0264-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-010-0264-2

Keywords

Search

Navigation