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Protein Precipitation to Remove Carbohydrates that Interfere in Protein-Bound Tryptophan Quantification in Soybean Seeds

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Journal of Analysis and Testing 2020年第3期4卷 238-247页

作者： Zhentian Lei Santosh Kumar Saurav J.Sarma Ritesh Kumar barbara w.sumner Minviluz G.Stacey Lloyd w.sumner University of Missouri Metabolomics Center ColumbiaMO 65211USA Division of Biochemistry University of MissouriColumbiaMO 65211USA Division of Plant Sciences University of MissouriColumbiaMO 65211USA

Tryptophan is one of the nine essential amino acids in humans that can only be obtained through diets and *** is a precursor to many biological processes,such as serotonin,melatonin,kynurenin,and niacin(nicotinamide)vitamin *** content of tryptophan in foods,such as soybean is an important indicator of nutritional ***,accurate quantification of tryptophan in soybean is crucial to soybean nutritional *** of soybean protein-bound amino acids first involves acid hydrolysis of total protein to liberate amino ***,tryptophan quantification following acid hydrolysis is difficult or impossible due to its reactions with soybean ***,removal of carbohydrates from soy proteins prior to acid hydrolysis is *** this study,we compared four common protein precipitation methods(i.e.,methanol,acetonitrile,acetone,and trichloroacetic acid(TCA)protein precipitation methods)to determine the best method to separate soy proteins from carbohydrates,and concluded that acetone provided the highest recovery of soy *** content in the precipitated proteins was determined after acid hydrolysis of the proteins using liquid chromatography-tandem mass spectrometry multiple reaction monitoring(LC-MS/MS-MRM).No significant difference in the tryptophan content was found among proteins precipitated with methanol,acetonitrile,and TCA,suggesting that these precipitated proteins have similar compositions.A slightly lower,but statistically significant tryptophan content was found in the acetonitrile-precipitated proteins,suggesting that these proteins contain slightly higher glycosylated proteins.

关键词： Tryptophan Acid hydrolysis LC-MS/MS MRM Soybean seeds Protein precipitation

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