Solubilized substrates for the on-line measurement of lipases by flow injection analysis during chromatographic enzyme purification.
A flow injection analysis (FIA) system for the on-line measurement of lipases in chromatographic processes has been developed. The photometrically detectable substrates para-nitrophenylpalmitate, S,O,O'-tripropyryl-1-thioglycerol, and 1,2-O-dilauryl-rac-glycero-3-glutaric-resorufinester were investigated. Different detergents and qualities of assay emulsions were tested for optimal results in FIA applications. Emphasis was placed on increasing the stability of the assay emulsion. Lipases of different origin and specificity were detected. The linear detection range was adapted to the requirements of the chromatographic purification procedures. The connection of the FIA with a fast protein liquid chromatography system permitted the automatization of lipase purification by monitoring protein content, salinity, and enzyme activity of the effluent from column chromatography.
Chemnitius
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GC
GBF--Gesellschaft für Biotechnologische Forschung mbH, Department of Enzyme Technology, Braunschweig, Germany.
Erdmann
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Schmid
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D000818:Animals; D002853:Chromatography, Liquid; D004075:Diglycerides / Q000378:metabolism*; D017022:Flow Injection Analysis; D005995:Glyceryl Ethers / Q000378:metabolism*; D008049:Lipase / Q000302:isolation & purification / Q000378:metabolism*; D010078:Oxazines / Q000378:metabolism*; D010168:Palmitates / Q000378:metabolism*; D013440:Sulfides / Q000378:metabolism*
D004075:Diglycerides; D005995:Glyceryl Ethers; D010078:Oxazines; D010168:Palmitates; D013440:Sulfides; C075376:resorufinyl 1,2-O-dilaurylglycero-3-glutarate; C075371:S,O,O'-tripropyryl-1-thioglycerol; C050015:4-nitrophenyl palmitate; D008049:Lipase