Cross-linked crystals of chloroperoxidase.
Chloroperoxidase from Caldariomyces fumago was crystallized. The crystals were modified with several cross-linkers, but only glurataldehyde was able to produce catalytically active and insoluble crystals. Unlike other immobilized chloroperoxidase preparations, these catalytic crystals are more thermostable than the unmodified soluble enzyme. The enhanced stability is probably due to the structure conservation in the crystalline matrix. In addition, non-cross-linked chloroperoxidase crystals retained more activity than the soluble enzyme after incubation in an organic solvent with low water content. Although the cross-linked crystals were catalytically active, they showed lower specific activity than the soluble enzyme. This low activity may be due to non-specific reactions between the cross-linker and essential residues for catalysis. Alternative cross-linking strategies are discussed.
Institute of Biotechnology, UNAM. Apartado Postal 510-3, Cuernavaca, Morelos, Mexico. maa@ibt.unam.mx
Marcela
M
Ayala
Eduardo
E
Horjales
Michael A
MA
Pickard
Rafael
R
Vazquez-Duhalt
D002384:Catalysis; D002711:Chloride Peroxidase / Q000737:chemistry* / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000494:pharmacology; D003460:Crystallization; D005658:Fungi / Q000201:enzymology; D005976:Glutaral / Q000737:chemistry; D006861:Hydrogen Peroxide / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D011485:Protein Binding; D013696:Temperature; D014867:Water / Q000737:chemistry
D003432:Cross-Linking Reagents; D014867:Water; D006861:Hydrogen Peroxide; D002711:Chloride Peroxidase; D005976:Glutaral