Enzymatic assay method for evaluating the lipase activity in complex extracts from transgenic corn seed.
A colorimetric method was established to determine the activity of recombinant lipase in extracts from transgenic corn seed. The system was an oil-in-water emulsion that was stabilized by a surfactant to accommodate the organic phase substrate and aqueous phase enzyme. The lipase activity was measured by monitoring the release of nitrophenol at 346 nm from the substrate, 4-nitrophenyl butyrate. Emulsions prepared with various surfactant types and concentrations were tested. For each surfactant, the measured activity was greatest when the surfactant concentration was close to the critical micelle concentration, consistent with the changing trend of oil droplet size as a function of surfactant concentration. The optimal system, with 0.01% (w/w) Tween 80, demonstrated good reproducibility, high sensitivity, robustness, and a linear response to lipase concentration.
Zhong
Qixin
Q
Department of Chemical and Biological Engineering, Iowa State University, Ames, Iowa 50011, USA. qzhong@utk.edu
Glatz
Charles E
CE
D003124:Colorimetry / Q000379:methods*; D004655:Emulsions; D006863:Hydrogen-Ion Concentration; D008049:Lipase / Q000032:analysis* / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000201:enzymology*; D011994:Recombinant Proteins / Q000032:analysis; D012639:Seeds / Q000201:enzymology*; D003313:Zea mays / Q000235:genetics*
D004655:Emulsions; D011994:Recombinant Proteins; D008049:Lipase