ABSTRACT
White-rot fungus manganese peroxidase (MnP) oxidizes a wide range of substrates, rendering it an interesting enzyme for potential applications. The stability of MnP can be improved by immobilization. With sodium alginate, gelatin, or chitosan as a carrier, and glutaraldehyde as the crosslinking agent, MnP was co-immobilized using the embed-crosslinked method and the adsorb-crosslinked method. The immobilization conditions and the partial properties of the three immobilized enzymes were investigated. When compared with the free enzyme, the optimum pH values and the temperatures of the three immobilized MnPs carried by alginate, gelatin, and chitosan were respectively shifted from 7.0 to 5.0, 5.0, 3.0 and from 35 degrees C to 75 degrees C , 55 degrees , 75 degrees C . The thermostabilities of the three immobilized MnPs were considerably better than that of the native enzyme. The chitosan-decreased by less than 5% even after repeated use for 6 - 9 times. The ability of decolorizing azo dyes in static and shaky situation by gelatin-immobilized MnP approached to the free enzyme, and there was no loss of enzyme activity during 2 repeated batch reactions.
Subject(s)
Adsorption , Alginates , Chemistry , Metabolism , Biocatalysis , Chitosan , Chemistry , Metabolism , Dose-Response Relationship, Drug , Enzymes, Immobilized , Chemistry , Metabolism , Fungal Proteins , Chemistry , Metabolism , Gelatin , Chemistry , Metabolism , Glucuronic Acid , Chemistry , Metabolism , Glutaral , Pharmacology , Hexuronic Acids , Chemistry , Metabolism , Hydrogen-Ion Concentration , Kinetics , Peroxidases , Chemistry , Metabolism , Schizophyllum , Substrate Specificity , TemperatureABSTRACT
<p><b>AIM</b>The process of vascular calcification involves various genetic alterations which may play a very important role in the vascular calcification. Vascular smooth muscle cells undoubtedly composed the main part of vascular cells, and are involved in vascular calcification. So bovine artery smooth muscle cell (BASMC) was used to investigate the gene changes during BASMC's calcification.</p><p><b>METHODS</b>Bovine artery smooth muscle cells cultured in vitro was induced calcified by beta-Glycerophosphate (beta-GP). Using DD-PCR technique to screening differentially expressed genes and those differentially expressed bands were reexamined by reverse Northern blot. All the ESTs were sequenced and BLAST with GenBank.</p><p><b>RESULTS</b>Total 65 cDNAs were isolated as differentially expressed genes and 40 of them were successfully reamplified. Using reverse-Northern blot, seven of these 40 cDNAs were reproducibly expressed differentially between the two cells. Three of them are new bands and have not been reported before.</p><p><b>CONCLUSION</b>This is the first time using DD-PCR to screen differentially expressed genes of BASMC calcification. Seven related ESTs were identified relating to BASMC calcification.</p>