化学
硫醇
试剂
半胱氨酸
吸光度
十二烷基硫酸钠
色谱法
布拉德福德蛋白质测定
有机化学
酶
作者
Kin-Yu Chan,Bruce P. Wasserman
出处
期刊:Cereal Chemistry
日期:1993-02-01
卷期号:70 (1): 22-26
被引量:133
摘要
A direct colorimetric method that simultaneously combines measurement of solubilized and insoluble thiol groups and disulfide bonds in corn meal-based materials is described. Ellman's reagent, 5,5'-dithiobis (2-nitrobenzoic acid), which reacts specifically with thiol groups, or disodium 2-nitro-5-thiosulfobenzoate, which reacts with cysteine and thiol groups formed after reduction of disulfide bonds with sodium sulfite, were reacted directly with corn meal in the presence of surfactants (urea and/or sodium dodecyl sulfate), releasing the soluble chromophore 2-nitro-5-thiobenzoate. After a clarification step to remove suspended material, absorbance at 412 nm was read. This assay was highly reproducible, and measurements agreed with direct amino acid analysis. Twin-screw extrusion of corn meal at 150 degrees C at moisture levels of 16 and 18% had no significant effect on cysteine or disulfide bond levels. Other possible changes such as disulfide bond rearrangements could not be determined by the mixed-phase assay. This method provides a rapid and convenient means for screening thiol and disulfide levels in insoluble proteinaceous materials
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