Cross-linking and immobilisation of different proteins with recombinant Verrucomicrobium spinosum tyrosinase.
This paper reports on the cross-linking and immobilisation of various proteins by the recombinant tyrosinase from Verrucomicrobium spinosum (Vs-tyrosinase). In general it is found that Vs-tyrosinase can readily cross-link proteins with a low degree of complexity, such as casein, but that the enzyme cannot readily cross-link well folded protein substrates such as lysozyme, myoglobin, cytochrome c or Candida antarctica lipase B (CALB). However, the inclusion of phenolic compounds (phenol or caffeic acid) to reaction mixtures of these proteins can greatly enhance the levels of cross-linking. For example it is possible to prepare cross-linked aggregates of industrially applicable enzymes such as CALB by simply incubating it with Vs-tyrosinase and phenol. The resulting aggregates can be collected by centrifugation and retain high levels of activity and may find applications in biocatalysis.
Fairhead
Michael
M
EMPA, Swiss Federal Laboratories for Materials Testing and Research, Laboratory for Biomaterials, Lerchenfeldstrasse 5, St. Gallen, CH-9014, Switzerland.
Thöny-Meyer
Linda
L
D001419:Bacteria / Q000201:enzymology*; D055162:Biocatalysis; D002364:Caseins / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000378:metabolism*; D045304:Cytochromes c / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins; D055767:Immobilized Proteins / Q000378:metabolism*; D008049:Lipase / Q000378:metabolism*; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D009113:Muramidase / Q000378:metabolism; D009211:Myoglobin / Q000378:metabolism; D019800:Phenol; D011994:Recombinant Proteins / Q000378:metabolism*
D002364:Caseins; D003432:Cross-Linking Reagents; D005656:Fungal Proteins; D055767:Immobilized Proteins; D009211:Myoglobin; D011994:Recombinant Proteins; D019800:Phenol; D045304:Cytochromes c; D014442:Monophenol Monooxygenase; D008049:Lipase; C431349:lipase B, Candida antarctica; D009113:Muramidase