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Optimizing Assay and Extraction of Lipoxygenase in Wheat Germ
Author(s) -
BHIRUD PRAKASH R.,
SOSULSKI F.W.,
SOSULSKI K.
Publication year - 1993
Publication title -
journal of food science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.772
H-Index - 150
eISSN - 1750-3841
pISSN - 0022-1147
DOI - 10.1111/j.1365-2621.1993.tb06121.x
Subject(s) - chemistry , ionic strength , extraction (chemistry) , chromatography , linoleic acid , lipoxygenase , substrate (aquarium) , pulmonary surfactant , wheat germ , ethanol , phosphate buffered saline , phosphate , enzyme , biochemistry , organic chemistry , fatty acid , oceanography , aqueous solution , geology
Using three‐level seven‐factor response surface methodology, wheat germ lipoxygenase (LPO) assay conditions were standardized. The important parameters were concentration of the substrate (linoleic acid), and surfactant (Tween 20), pH and temperature. The standardized LPO assay conditions for the 1 mL reaction volume were : 450 μM linoleic acid, 129 μM Tween 20, 175 mM ethanol, 1 mM EDTA, pH 6.2 (phosphate buffer), ionic strength 100 mM and 40°C. LPO extraction conditions were standardized by sequential variation of parameters. Optimum conditions were a single extraction of defatted wheat germ flour at 2–5°C with magnetic stirring of an extractant acetate buffer pH 4.5, ionic strength 100 mM, at buffer‐to‐solid ratio 10:l.