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The impact of Fusarium culmorum infection on the protein fractions of raw barley and malted grains

机译:镰刀菌感染对生大麦和麦芽谷物蛋白质组分的影响

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摘要

Contaminating fungi, such as Fusarium species, produce metabolites that may interfere with normal barley grain proteolysis pattern and consequently, affect malt and beer quality. Protein compositional changes of an initial mixture of 20 % Fusarium culmorum infected and 80 % noninfected mature barley grains and respective malt are reported here. Proteolytic activity of infected barley grains (IBG) and respective malt, with controls (uninfected grains), were characterized using protease inhibitors from each class of this enzyme, including metallo-, cysteine, serine, and aspartic proteases, as well as uninhibited protease fractions. The proteins were extracted according to the Osborne fractionation and separated by size exclusion chromatography. Additionally, two-dimensional (2D) gel electrophoresis (GE) was used to analyze hydrophobic storage proteins isolated from the control and IBG. Analyses revealed that F. culmorum IBG had a twofold increase of proteolytic activity compared to the control sample, which showed an increase in all protease classes with aspartic proteases dominating. Infected and control malt grains were comparable with cysteine proteases representing almost 50 % of all proteolytic enzymes detected. Protein extractability was 31 % higher in IBG compared to the control barley. The albumin fraction showed that several metabolic proteins decreased and increased at different rates during infection and malting, thus showing a complex F. culmorum infection interdependence. Prolamin storage proteins were more hydrophobic during barley fungal infection. F. culmorum interfered with the grain hydrolytic protein profile, thereby altering the grain's protein content and quality.
机译:诸如镰刀菌(Fusarium)等污染性真菌产生的代谢产物可能会干扰正常的大麦籽粒蛋白水解模式,从而影响麦芽和啤酒的品质。此处报告了感染了20%镰刀菌和80%未感染的成熟大麦籽粒和相应麦芽的初始混合物的蛋白质组成变化。使用来自该酶每一类的蛋白酶抑制剂(包括金属蛋白酶,半胱氨酸,丝氨酸和天冬氨酸蛋白酶以及未抑制的蛋白酶馏分)来表征感染的大麦籽粒(IBG)和相应的麦芽以及对照(未感染的籽粒)的蛋白水解活性。 。根据奥斯本分级分离法提取蛋白质,并通过尺寸排阻色谱法分离。此外,二维(2D)凝胶电泳(GE)用于分析从对照和IBG中分离的疏水性存储蛋白。分析显示,与对照样品相比,F。culmorum IBG的蛋白水解活性增加了两倍,这表明所有蛋白酶类别均以天冬氨酸蛋白酶为主增加。感染和对照的麦芽谷物与半胱氨酸蛋白酶相当,半胱氨酸蛋白酶几乎占所有检测到的蛋白水解酶的50%。与对照大麦相比,IBG中的蛋白质提取能力高31%。白蛋白级分显示,在感染和麦芽形成过程中,几种代谢蛋白以不同的速率减少和增加,因此显示出复杂的鳞茎镰刀菌感染相互依赖。谷蛋白贮藏蛋白在大麦真菌感染过程中更具疏水性。 F. culmorum干扰了谷物的水解蛋白质谱,从而改变了谷物的蛋白质含量和质量。

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