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Redox-dependent interaction between thaumatin-like protein and β-glucan influences malting quality of barley

机译:索马甜蛋白样蛋白与β-葡聚糖之间的氧化还原依赖性相互作用影响大麦的发芽品质

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摘要

Barley is the cornerstone of the malting and brewing industry. It is known that 250 quantitative trait loci (QTLs) of the grain are associated with 19 malting-quality phenotypes. However, only a few of the contributing genetic components have been identified. One of these, on chromosome 4H, contains a major malting QTL, QTL2, located near the telomeric region that accounts, respectively, for 28.9% and 37.6% of the variation in the β-glucan and extract fractions of malt. In the current study, we dissected the QTL2 region using an expression- and microsynteny-based approach. From a set of 22 expressed sequence tags expressed in seeds at the malting stage, we identified a candidate gene, TLP8 (thaumatin-like protein 8), which was differentially expressed and influenced malting quality. Transcript abundance and protein profiles of TLP8 were studied in different malt and feed varieties using quantitative PCR, immunoblotting, and enzyme-linked immunosorbent assay (ELISA). The experiments demonstrated that TLP8 binds to insoluble (1, 3, 1, 4)-β-D glucan in grain extracts, thereby facilitating the removal of this undesirable polysaccharide during malting. Further, the binding of TLP8 to β-glucan was dependent on redox. These findings represent a stride forward in our understanding of the malting process and provide a foundation for future improvements in the final beer-making process.
机译:大麦是制麦和酿造业的基石。已知谷物的250个数量性状基因座(QTL)与19个麦芽品质表型相关。但是,仅鉴定了一些促成遗传成分。其中之一,在4H染色体上,包含一个主要的麦芽QTL QTL2,位于端粒区域附近,分别占β-葡聚糖和麦芽提取物部分变异的28.9%和37.6%。在当前的研究中,我们使用基于表达和微同步的方法解剖QTL2区域。从一组在发芽阶段的种子中表达的22个表达序列标签中,我们鉴定了一个候选基因TLP8(索马甜蛋白8),该基因差异表达并影响了发芽质量。使用定量PCR,免疫印迹和酶联免疫吸附测定(ELISA),研究了不同麦芽和饲料品种中TLP8的转录本丰度和蛋白质谱。实验表明,TLP8与谷物提取物中的不溶性(1、3、1、4)-β-D葡聚糖结合,从而有助于在麦芽制造过程中去除这种不良多糖。此外,TLP8与β-葡聚糖的结合依赖于氧化还原。这些发现代表了我们对麦芽制造过程的理解的一大进步,并为将来啤酒最终制造过程的改进奠定了基础。

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