Oxidation of peptides and proteins by Trichoderma reesei and Agaricus bisporus tyrosinases.
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 beta-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 beta-casein could be cross-linked by TrTyr already in three hours, but large and compact BSA was not cross-linked even in 24h. 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.
Mattinen
Maija-Liisa
ML
VTT Technical Research Centre of Finland, P.O. Box 1000, FIN-02044 VTT, Finland. Maija.Mattinen@vtt.fi
Lantto
Raija
R
Selinheimo
Emilia
E
Kruus
Kristiina
K
Buchert
Johanna
J
D000364:Agaricus / Q000201:enzymology*; D000595:Amino Acid Sequence; D000818:Animals; D002364:Caseins / Q000737:chemistry / Q000378:metabolism*; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014242:Trichoderma / Q000201:enzymology*; D014443:Tyrosine / Q000378:metabolism; D014935:Wool / Q000378:metabolism
D002364:Caseins; D010455:Peptides; D012710:Serum Albumin, Bovine; D014443:Tyrosine; D014442:Monophenol Monooxygenase