Oxidation of peptides and proteins by Trichoderma reesei and Agaricus bisporus tyrosinases

• Published in: Journal of biotechnology 2008-02, Vol.133 (3), p.395-402
• Main Authors: Mattinen, Maija-Liisa, Lantto, Raija, Selinheimo, Emilia, Kruus, Kristiina, Buchert, Johanna
• Format: Article
• Language: English
• Subjects: Agaricus - enzymology; Agaricus bisporus; Amino Acid Sequence; Animals; Biological and medical sciences; Biotechnology; Caseins - chemistry; Caseins - metabolism; Cattle; Chromatography, High Pressure Liquid; Cross-linking; crosslinking; Electrophoresis, Polyacrylamide Gel; enzyme activity; Fundamental and applied biological sciences. Psychology; Grafting; Hypocrea jecorina; Microscopy, Fluorescence; Molecular Sequence Data; monophenol monooxygenase; Monophenol Monooxygenase - metabolism; oxidation; Oxidation-Reduction; oxidative crosslinking; Oxygen Consumption; Peptide; peptides; Peptides - chemistry; Peptides - metabolism; Protein; proteins; Serum Albumin, Bovine - chemistry; Serum Albumin, Bovine - metabolism; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Trichoderma - enzymology; Tyrosinase; Tyrosine; Tyrosine - metabolism; Wool - metabolism
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2007.10.009

The capability of a novel tyrosinase from Trichoderma reesei (TrTyr) to catalyse the oxidation and oxidative cross-linking of l-tyrosine ( l-Y) and tyrosine side-chains in GYG and EGVYVHPV peptides, in bovine serum albumin (BSA) and β-casein proteins as well as in proteinaceous wool fibres was studied by oxygen consumption measurement, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), reverse phase high-performance liquid chromatography (RP-HPLC), matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and fluorescence microscopy. TrTyr was compared to the well-characterised tyrosinase from Agaricus bisporus (AbTyr) in terms of oxidation and cross-linking. According to the results obtained TrTyr was capable of cross-linking peptides and proteins more efficiently than AbTyr. However, the size and three-dimensional structure of the proteinaceous substrates proved to be crucial for the success of the enzymatic catalysis. Random coil β-casein could be cross-linked by TrTyr already in three hours, but large and compact BSA was not cross-linked even in 24 h. TrTyr could also be used to incorporate a diphenolic compound, l-dihydroxyphenyl alanine ( l-dopa), into protein fibres whereas incorporation of a monophenol, l-Y was less efficient. Thus TrTyr is a potential tool for protein cross-linking and/or modification.