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技术说明:使用比浊法定量分析玉米、高水分玉米和玉米青贮中的醇溶蛋白:异丙醇和叔丁醇的比较效率。

Technical note: quantification of zeins from corn, high-moisture corn, and corn silage using a turbidimetric method: comparative efficiencies of isopropyl and tert-butyl alcohols.

机构信息

Feed and Food Science and Nutrition Institute, Faculty of Agriculture, Università Cattolica del Sacro Cuore, 29100 Piacenza, Italy.

出版信息

J Dairy Sci. 2012 Jun;95(6):3384-9. doi: 10.3168/jds.2011-4995.

Abstract

Zeins are corn endosperm storage proteins that encapsulate starch granules into a protein matrix, which can act as a barrier to starch accessibility and digestion. Laboratory methods to quantify zein are seldom used because they are considered arduous and time-consuming. A recently published rapid turbidimetric method (mTM) was reinvestigated by changing the solution originally used for the zein solubilization step. In particular, the aim was to explore whether, and to what extent, the use of tert-butyl alcohol (t-BuOH-mTM) in lieu of isopropyl alcohol (i-PrOH-mTM) was able to improve the quantification of zeins from dry corn, high-moisture corn, and corn silage samples. The nature of the alcohol influenced the zein extraction values, and t-BuOH-mTM gave higher zein values in corn (3.6 vs. 3.3 g/100 g of dry matter) and corn silage samples (1.2 vs. 0.9 g/100 g of dry matter) compared with i-PrOH-mTM. In contrast, similar zein extraction values were obtained for high-moisture corn (2.1 vs. 1.9 g/100 g of dry matter, respectively). Sodium dodecyl sulfate-PAGE analysis revealed no contamination by nonzein proteins with the use of tert-butyl alcohol. Overall, these findings indicated that tert-butyl alcohol has a greater ability to solubilize zein compared with isopropyl alcohol and thus the t-BuOH-mTM allowed greater extraction of zeins. Considering the growing interest of animal nutritionists in zein proteins, such results should provide useful information for routine laboratory analysis.

摘要

玉米醇溶蛋白是玉米胚乳的储存蛋白,可将淀粉颗粒包裹在蛋白质基质中,从而起到阻碍淀粉消化的作用。由于实验室定量玉米醇溶蛋白的方法较为繁琐、耗时,所以很少使用。最近发表的快速浊度法(mTM)对玉米醇溶蛋白的溶解步骤中所使用的溶液进行了重新研究。具体而言,本研究旨在探讨是否以及在何种程度上,用叔丁醇(t-BuOH-mTM)代替异丙醇(i-PrOH-mTM)可以提高对干玉米、高水分玉米和青贮玉米样品中玉米醇溶蛋白的定量分析。醇的性质会影响玉米醇溶蛋白的提取值,与 i-PrOH-mTM 相比,t-BuOH-mTM 使玉米(3.6 比 3.3 g/100 g 干物质)和青贮玉米样品(1.2 比 0.9 g/100 g 干物质)中的玉米醇溶蛋白值更高。相比之下,高水分玉米中的玉米醇溶蛋白提取值相似(2.1 比 1.9 g/100 g 干物质)。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析表明,使用叔丁醇时没有非醇溶蛋白污染。总的来说,这些发现表明,与异丙醇相比,叔丁醇具有更强的溶解玉米醇溶蛋白的能力,因此 t-BuOH-mTM 可以更好地提取玉米醇溶蛋白。鉴于动物营养学家对玉米醇溶蛋白的兴趣日益浓厚,这些结果应为常规实验室分析提供有用信息。

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