結合酵素-層析法分析可食用菇類中具分支之(1,3; 1,6)-b-D-葡萄聚醣含量

Branched (1,3; 1,6)-b-D-glucan content in edible mushrooms: Enzymatic-chromatographic determination

黃琬庭(Wan-Ting Huang)

指導教授 ： 呂廷璋

國立臺灣大學/生物資源暨農學院/食品科技研究所/碩士(2013年)

https://doi.org/10.6342/NTU.2013.10715

摘要

菇蕈類食品及相關保健食品，其具有 (1,6)-β-葡萄糖基之 (1,3)-β-D-葡萄聚醣為已知的生物活性調節劑 (biological response modifier)，且已經發展為腫瘤抑制劑之純化藥劑的生產，此多醣為一活性成分指標。本研究選擇酵素-層析法進行分析方法的確校試驗，評估分析方法的精確性、準確性、專一性，並利用洋菇為代表性基質進行基質干擾評估，並將台灣常見的22種菇蕈類做 (1,3; 1,6)-b-D-葡萄聚醣含量及分支度分析。最後將針對另一重組酵素做其活性、專一性及轉換率試驗。實驗結果顯示所評估的分析方法於連續五天每天分析五次的精確度 (precision, % CV) 為5%，葡萄糖單體與雙體的標準線性分別為r2=0.9985與0.9991；檢測兩個年份的菇蕈類，其檢測結果為5.25 (mg/g)~207.53 (mg/g) 之間，而分支度在0.11~0.45之間，以市售的菇蕈類產品其含量範圍在9.89~54.79 (mg/g)。並以0.5 M及2 M的鹼相比較對於八種菇蕈類樣品的 (1,3; 1,6)-b-D-葡聚醣其含量及分支度比較，發現質地較軟的菇類樣品使用0.5 M的鹼溶即可達到使用2 M鹼溶的效果，但如為質地較硬的樣品則仍需使用2 M的鹼溶。對於recombinant exo-1,3-β-D-glucanase與endo-1,3-β-D-glucanase其混和使用的合適環境為40℃，及合併使用後對於非菇蕈類基質的作用，以整體而言，recombinant exo-1,3-β-D-glucanase對於β-(1,3)葡聚醣的基質較為專一。

關鍵字

(1,3 ； 1,6)-β-D-葡聚醣 ； 可食用菇類 ； exo-1,3-β-D-glucanase ； endo-1,3-β-D-glucanase

並列摘要

Edible mushrooms and their products have (1,6)-β-branched (1,3)-β-D-glucans, which play a targeting biological function as biological response modifiers, and have been developed by the pharmaceutical industry as a tumor inhibitor. This study employed enzymatic-chromatographic determination method to do validation, including precision, accuracy, specific, and matrix addition, and analysis of (1,3; 1,6)-b-D-glucan content and degree of branching for 22 species of Taiwan cultivated edible mushrooms. Then aim to recombinant exo-1,3-β-D-glucanase ( recombinant exo-1,3-ß-glucanase) to observe activity, specific, and conversion test. Samples were analyzed five times a day for five days. Results showed a precision (% CV) of 5%, with R square value 0.9985 and 0.9991 for glucose and gentiobiose, respectively. Samples of two different age mushrooms had a (1,3;1,6)-b-D-glucan content of 5.25~207.53 (mg/g), and degree of branching of 0.11~0.45. On the other hand, commercial mushroom products have 9.89-54.79 (mg/g) (1,3;1,6)-b-D-glucan content. Two alkali concentration 0.5M and 2M were used to analyze eight samples for its (1,3; 1,6)-b-D-glucan content and degree of branching. For soft textured samples, both 0.5M and 2M alkali concentration could be used and had the same effect. but for hard texture samples, 2M alkali concentration must be used for recombinant exo-1,3-β-D-glucanase mixing endo-1,3-β-D-glucanase the optimum temperature is 40℃, and action the non-(1,3; 1,6)-b-D-glucan matrix. Overall, recombinant exo-1,3-β-D-glucanase is specific of β-(1,3)-D-glucan sample, but if the specific of β-(1,3)-D-glucan sample.

並列關鍵字

(1,3 ； 1,6)-b-D-glucan ； edible mushrooms ； exo-1,3-β-D-glucanase ； and endo-1,3-β-D-glucanase

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