Continuous aqueous phase extraction of proteins: automated on-line monitoring of fumarase activity and protein concentration.
Automated assay techniques are described for on-line measurements of fumarase activity and total protein concentration, including in-line sample dilution and sample multiplexing during continuous aqueous phase extraction. Fumarase was determined by following the conversion of L-malate to fumurate at a wavelength of 250 nm, while the protein assay was based on the Biuret reaction. Actual assay times of 2 and 4 min were achieved for the fumarase and protein measurements, respectively, with an effective measurement cycle time of 2 min. Standard deviations of c. 3.2 and 2% of the measured values were calculated for the enzyme and protein values, respectively. The assay system was coupled to a computer to allow on-line data visualization and storage.
Papamichael
N
N
GBF, Gesellschaft für biotechnologische Forschung, Braunschweig, FRG.
Börner
B
B
Hustedt
H
H
D001331:Automation; D002498:Centrifugation; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation*; D003626:Data Display; D005649:Fumarate Hydratase / Q000032:analysis* / Q000302:isolation & purification; D005656:Fungal Proteins / Q000032:analysis* / Q000302:isolation & purification; D009862:Online Systems; D012997:Solvents; D014867:Water
D005656:Fungal Proteins; D012997:Solvents; D014867:Water; D005649:Fumarate Hydratase