- 學位論文
麥芽用大麥胚乳澱粉粒結合蛋白的研究
Studies on the starch granule-associated proteins in malting barley endosperm
摘要
本論文的研究內容主要是針對四種不同麥芽特性分類的15個蘇格蘭大麥品種，利用蛋白質體學 (proteomics) 來研究其胚乳的澱粉顆粒結合蛋白 (starch granule-associated protein)。其中麥芽特性最佳的8個品種是目前蘇格蘭威士忌酒的釀酒品種，其餘則包括舊釀酒 品種、麥芽特性差品種及飼料用品種。 我們將四種大麥品種分別進行澱粉粒表面及嵌合蛋白之SDS- PAGE電泳，結果顯示表面蛋白多集中於30-45 kDa之間，嵌合蛋白則大多集中於25-100 kDa之間，且以分子量約94 kDa、86 kDa、74 kDa、62 kDa四個條帶最為清晰。此外，我們分別將15個蘇格蘭大麥品種的澱粉顆粒結合蛋白進行雙向電泳，並將圖譜分成五個區域作分析及探討。所有品種在區域1呈現的圖譜差異不大，並皆可明顯看到starch synthase I (SS I) 及 granule-bound starch synthase I (GBSS I) 兩蛋白。區域2大約位於分子量50 kDa附近，pH 5-6.5之間，大部分的品種都明顯具有編號21、22、23的點，我們初步認為此區蛋白呈現的差異，不是影響釀酒特性的關鍵蛋白。區域3的蛋白大多為serpin Z4蛋白，我們發現非釀酒品種在serpin Z4的量上有較少的趨勢，所以serpin Z4蛋白的含量與篩選出釀酒品種呈現部分正相關。區域4的蛋白主要是屬於大麥儲藏性蛋白中的B hordein，而飼料用品種在此區出現部分特異圖譜，顯示此圖譜可初步將飼料用品種區分出來。區域5的蛋白與分出麥芽特性較好的品種有某程度的正相關，但針對其中強度較高的蛋白質點進行分析後，未能得到符合的蛋白身分，因此這將是本實驗未來研究方向的重點之一。
並列摘要
In the present study, we analyzed starch granule-associated proteins by using proteomics from fifteen kinds of Scotland barley varieties which were classified into four levels of malt quality. For the best 8 varieties of malts, they were varieties for brewing by the Scotland Whisky factories in the present time. Besides, the old varieties for brewing, the varieties with bad malt characteristic and the varieties for the feed were also used. First, we studied on those four kinds of barley varieties by regular one dimension SDS-PAGE on starch granule surface and integral proteins. The results indicated starch granule surface proteins were mainly between 30-45 kDa while integral proteins distributed between 25-100 kDa with four main protein bands, i.e. the 94 kDa, 86 kDa, 74 kDa and 62 kDa proteins. For further analysis, these proteins were analyzed by two-dimensional electrophoresis and we divided the protein spots into five regions for the convinience of detail analysis. The difference of these varieties at region 1 which contained starch synthase I (SS I) and granule-bound starch synthase I (GBSS I) were not very obvious. The region 2 located on the area with molecular weight about 50 kDa and between pH5~6.5. Most of these varieties contained number 21, 22 and 23 spots. It was hypothesized that the difference of the proteins in this area should not be essential proteins for brewing characteristics. The region 3 proteins were mostly serpin Z4 and the proteins in the non-brewing varieties contained lower amount of serpin Z4. Thus, there were positive relationship between the amount of serpin Z4 and the characteristic of the barley brewing ability. The proteins in region 4 were mainly hordein B, one kind of the barley storage proteins. The feed varieties gave different patterns in this area which might differentiate the varieties for feeding from other varieties. The proteins in region 5 would also showed positive correlation with the characteristic for brewing ability, however, we could not recognize the proteins identity in this region yet. Further analysis should be perform in order to understand the relationship between these proteins and brewing ability.
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