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首页> 外文期刊>Cereal Chemistry >Improved methods for high-throughput extraction and assay of green barley malt proteinase activity facilitating examination of proteinase activity across large-scale barley populations.
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Improved methods for high-throughput extraction and assay of green barley malt proteinase activity facilitating examination of proteinase activity across large-scale barley populations.

机译：高通量提取和测定绿色大麦麦芽蛋白酶活性的改进方法，有助于检查大麦大麦群体的蛋白酶活性。

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摘要

We report efficient sample extraction and assay methods allowing quantitative determinations of proteinase activities from barley malt. The improved methods are used to assay >2,200 developmental lines of malting barley for two subsets of proteinase activity. The distributions of the resulting activities suggest differences in population structures between the two types of proteinases. Comparison of the activities of the green malt proteinases with standard malting quality measurements show highly significant correlations that differ between the proteinase subsets. The pH 4.5 hydrolysis of the artificial substrate Z-Phe-Arg-AMC correlates well with the traditional malting quality measurements, supporting the role of cysteine-class proteinases in mobilization of grain reserves during malting and mashing. Results from assays of gelatin hydrolysis at pH 6.0 suggest that these proteolytic activities may be involved in other aspects of seed C and N dynamics also linked to malting quality measurements. The differences between the pH 4.5 and 6.0 activities assayed here and their association with malting quality measurements suggest different physiological roles for the two proteinase activities in several aspects of seed germination. Either assay could be useful for population surveys, depending on the particular facet of seed metabolism under study.
机译：我们报告了有效的样品提取和测定方法，可以定量测定大麦麦芽中的蛋白酶活性。改进的方法用于分析麦芽大麦的> 2,200个发育系的两个蛋白酶活性子集。所得活性的分布表明两种蛋白酶之间的种群结构存在差异。绿色麦芽蛋白酶的活性与标准麦芽品质测量结果的比较显示，蛋白酶子集之间存在高度显着的相关性。人造底物Z-Phe-Arg-AMC的pH 4.5水解与传统制麦芽质量测量密切相关，支持半胱氨酸类蛋白酶在制麦芽和糖化过程中动员谷物储备中的作用。在pH 6.0的明胶水解试验结果表明，这些蛋白水解活性可能参与了种子C和N动态的其他方面，这些动态也与麦芽质量测量有关。此处测定的pH 4.5和6.0活性之间的差异以及它们与麦芽品质测量的关联表明，在种子萌发的几个方面，两种蛋白酶活性的生理作用不同。取决于所研究的种子代谢的特定方面，这两种测定法均可用于种群调查。

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