Direct determination of the cephalosporin transforming activity of immobilized cells with use of an enzyme thermistor. 1. Verification of the mathematical model.
A simple method for the direct measurement of the catalytic properties of immobilized cells in the flow minicalorimeter, the enzyme thermistor (ET), is presented. A Trigonopsis variabilis strain with cephalosporin C-transforming activity was used as the model system. The yeast cells were immobilized either by crosslinking with a homobifunctional reagent or by entrapment in gels. The actual activity of the immobilized cells used in the ET was estimated by means of a stirred-batch reactor measurement in conjunction with HPLC analysis of substrate and products. Similar results were also obtained using D-amino acid oxidase (EC 1.4.3.3) isolated from T. variabilis cells and immobilized by gel entrapment. This calibration procedure was found to be appropriate for all biocatalyst systems used. The thermometric signal was proportional to the amount of biocatalyst immobilized in the ET minicolumn. It was shown that the rate of reaction catalyzed by T. variabilis entrapped in calcium pectate gel was limited by internal diffusion to an extent depending on the cell concentration in the biocatalyst particle. This approach offers a direct method for studying the kinetic properties of immobilized cells.
Gemeiner
P
P
Institute of Chemistry, Slovak Academy of Sciences, Bratislava.
Stefuca
V
V
Welwardová
A
A
Michálková
E
E
Welward
L
L
Kurillová
L
L
Danielsson
B
B
D000164:Acremonium / Q000378:metabolism*; D015374:Biosensing Techniques; D001709:Biotechnology / Q000379:methods; D002151:Calorimetry / Q000379:methods; D002511:Cephalosporins / Q000302:isolation & purification / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003432:Cross-Linking Reagents; D005976:Glutaral; D007202:Indicators and Reagents; D007700:Kinetics; D008433:Mathematics; D003904:Mitosporic Fungi / Q000378:metabolism*; D008962:Models, Theoretical; D010368:Pectins; D011094:Polyethyleneimine
D002511:Cephalosporins; D003432:Cross-Linking Reagents; D007202:Indicators and Reagents; C025163:cephalosporin C; D010368:Pectins; D011094:Polyethyleneimine; D005976:Glutaral; C003181:polygalacturonic acid