tailieunhanh - Báo cáo khoa học: Laccase-catalyzed polymerization of tyrosine-containing peptides

Laccase-catalyzed polymerization of tyrosine and tyrosine-containing pep-tides was studied in the presence and absence of ferulic acid (FA). Advanced spectroscopic methods such as MALDI-TOF MS, EPR, FTIR microscopy and HPLC-fluorescence, as well as more conventional analyt-ical tools: oxygen consumption measurements and SDS⁄PAGE were used in the reaction mechanism studies. Laccase was found to oxidize tyrosine and tyrosine-containing peptides, with consequent polymerization of the compounds. | iFEBS Journal Laccase-catalyzed polymerization of tyrosine-containing peptides Maija-Liisa Mattinen1 Kristiina Kruus1 Johanna Buchert1 Jacob H. Nielsen2 Henrik J. Andersen2 and Charlotte L. Steffensen2 1 VTT Biotechnology Espoo Finland 2 Danish Institute of AgriculturalSciences Department of Food Science Research Centre Foulum Tjele Denmark Keywords cross-linking ferulic acid laccase peptide tyrosine Correspondence . Mattinen VTT Biotechnology PO Box 1500 FIN-02044 VTT Tietotie 2 Espoo Finland Fax 358 20 7227071 Tel 358 20 7227143 E-mail Website http bel Received 15 March 2005 revised 18 May 2005 accepted 23 May 2005 Laccase-catalyzed polymerization of tyrosine and tyrosine-containing peptides was studied in the presence and absence of ferulic acid FA . Advanced spectroscopic methods such as MALDI-TOF MS EPR FTIR microscopy and HPLC-fluorescence as well as more conventional analytical tools oxygen consumption measurements and SDS PAGE were used in the reaction mechanism studies. Laccase was found to oxidize tyrosine and tyrosine-containing peptides with consequent polymerization of the compounds. The covalent linkage connecting the compounds was found to be an ether bond. Only small amounts of dityrosine bonds were detected in the polymers. When FA was added to the reaction mixtures it was found to be incorporated into the polymer structure. Thus in addition to homopolymers different heteropolymers containing two or four FA residues were formed in the reactions. doi Laccase EC is a multicopper enzyme belonging to the blue multicopper oxidase family. The most studied laccases are of fungal and plant origin 1-5 however some bacterial laccases 6 7 have also been isolated. In addition these enzymes have been found in some insects 8-10 . Laccases catalyze the oxidation of a wide range of organic and inorganic substances 11 . Typical organic substrates are aromatic compounds such as different

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