title: Trichoderma harzianum Produces a New Thermally Stable Acid Phosphatase, with Potential for Biotechnological Application.
abstract: Acid phosphatases (ACPases) are produced by a variety of fungi and have gained attention due their biotechnological potential in industrial, diagnosis and bioremediation processes. These enzymes play a specific role in scavenging, mobilization and acquisition of phosphate, enhancing soil fertility and plant growth. In this study, a new ACPase from Trichoderma harzianum, named ACPase II, was purified and characterized as a glycoprotein belonging to the acid phosphatase family. ACPase II presents an optimum pH and temperature of 3.8 and 65 °C, respectively, and is stable at 55 °C for 120 min, retaining 60% of its activity. The enzyme did not require metal divalent ions, but was inhibited by inorganic phosphate and tungstate. Affinity for several phosphate substrates was observed, including phytate, which is the major component of phosphorus in plant foods. The inhibition of ACPase II by tungstate and phosphate at different pH values is consistent with the inability of the substrate to occupy its active site due to electrostatic contacts that promote conformational changes, as indicated by fluorescence spectroscopy. A higher affinity for tungstate rather than phosphate at pH 4.0 was observed, in accordance with its highest inhibitory effect. Results indicate considerable biotechnological potential of the ACPase II in soil environments.
authors.affiliation: Laboratory of Biophysics, Department of Cellular Biology, University of Brasília, 70910-900, Brasília, Brazil.
authors.forename: Amanda Araújo
authors.initials: AA
authors.lastname: Souza
authors.affiliation: Laboratory of Enzymology, Department of Cellular Biology, University of Brasília, 70910-900, Brasília, Brazil.
authors.forename: Vanessa Oliveira
authors.initials: VO
authors.lastname: Leitão
authors.affiliation: Laboratory of Mass Espectrometry, Embrapa Recursos Genéticos e Biotecnologia - 70770-917, Brasília, Brazil.
authors.forename: Marcelo Henrique
authors.initials: MH
authors.lastname: Ramada
authors.affiliation: Laboratory of Biophysics, Department of Cellular Biology, University of Brasília, 70910-900, Brasília, Brazil.
authors.forename: Azadeh
authors.initials: A
authors.lastname: Mehdad
authors.affiliation: Laboratory of Enzymology, Institute of Biology, University Federal of Goiás, 74001-970, Goiania, Brazil.
authors.forename: Raphaela de Castro
authors.initials: Rde C
authors.lastname: Georg
authors.affiliation: Laboratory of Enzymology, Institute of Biology, University Federal of Goiás, 74001-970, Goiania, Brazil.
authors.forename: Cirano José
authors.initials: CJ
authors.lastname: Ulhôa
authors.affiliation: Laboratory of Biophysics, Department of Cellular Biology, University of Brasília, 70910-900, Brasília, Brazil.
authors.forename: Sonia Maria
authors.initials: SM
authors.lastname: de Freitas
mesh_terms: D000135:Acid Phosphatase / Q000096:biosynthesis* / Q000737:chemistry; D001709:Biotechnology / Q000379:methods*; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000737:chemistry; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D007218:Industrial Microbiology / Q000379:methods; D010710:Phosphates / Q000737:chemistry; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D055672:Static Electricity; D013696:Temperature; D014242:Trichoderma / Q000201:enzymology*; D017973:Tungsten Compounds / Q000737:chemistry
chemical_list: D005656:Fungal Proteins; D010710:Phosphates; D017973:Tungsten Compounds; D000135:Acid Phosphatase; C045951:tungstate