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論文名稱: | 蜈蚣藻乳酸發酵液之抗氧化性探討
| Studies on Antioxidation of Lactic Acid Fermented Grateloupia filicina
研究生: | 洪英豪 Ying-Hao Hung
指導教授: | 邱思魁 Tze-Kuei Chiou
| 潘崇良 Chorng-Liang Pan
學位類別: | 碩士
學校名稱: | 國立臺灣海洋大學
系所名稱: | 食品科學系
學號: | T92420021
畢業學年度: | 93
語文別: | 中文
關鍵字: | 蜈蚣藻 Grateloupia filicina
| 乳酸菌 lactic acid bacteria
| 抗氧化 Antioxidation
全文點閱次數: | 38

全文影像連結:

論文全文(pdf)

論文頁數: | 91
摘要: | 摘 要
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| 本研究是將市售蜈蚣藻 (Grateloupia filicina) 萃取多醣經α-amylase、cellulase、β-galactosidase、或由 Pseudomonas vesicularis MA103所產 agarase 等四種酵素水解後，挑出獲得寡單醣量較高的兩種酵素進行水解，以乳酸菌發酵該水解物，分析此乳酸發酵液之抗氧化能力。在相同酵素活性單位 (100 unit) 下水解蜈蚣藻多醣液，就水解後的還原醣量而言，cellulase 效果較佳，agarase 次之，且隨著酵素活性愈高，還原醣量愈高。
| 以 (A) 5% Lactobacillus (Lb.) rhamnosus BCRC14068、(B) 5% Enterococcus (Ent.) faecalis BCRC13076、與 (C) 2.5% Lb. rhamnosus BCRC14068 及 2.5% Ent. faecalis BCRC13076 於 37℃ 分別發酵 (1) 0.5% 蜈蚣藻多醣液經 agarase 及 cellulase (500 unit) 水解所得水解液及 (2) 該蜈蚣藻水解液添加 0.5% sucrose、1% beef extract 及 0.5% yeast extract (CN)，共得六組乳酸發酵液。結果顯示隨著發酵時間的增加，pH 值隨著下降。其中以 L-Gra-CN-FC5 組 Lb. rhamnosus BCRC14068 (2.5%) 及 Ent. faecalis BCRC13076 (2.5%) 發酵外加碳源與氮源的蜈蚣藻水解液反應至 9 hr 時，發酵液之 pH 值即可降至 4.6 以下，得一安全性產品，可滴定酸度為 0.74%，乳酸菌數 9.39 log CFU/mL。
| 六種乳酸發酵液在螯合亞鐵離子能力、血紅素催化亞麻油酸自氧化抑制率、還原力等抗氧化能力上，均以蜈蚣藻水解液另外添加碳氮源的基質發酵所得之發酵液較佳，除清除 DPPH 自由基能力。六種乳酸發酵液的可溶性多酚類含量，以 Lb. rhamnosus BCRC14068 及 Ent. faecalis BCRC13076 同時發酵外加碳源與氮源的蜈蚣藻水解液所含可溶性多酚類含量有 1,087.64 ± 0.31 GAE (gallic acid equivalents, μg/mL)，較其餘五種乳酸發酵液呈現較高多酚類含量 (400.27 ± 1.75 - 975.92 ± 4.90 GAE)。推測蜈蚣藻乳酸發酵液本身所含的胜肽、有機酸與可溶性多酚類等物質可能是提供抗氧化能力的成分。
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| Abstract
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| The polysaccharide extracts of Grateloupia (G.) filicina which was purchased from the market were digested by four enzymes, α-amylase, cellulase, β-galactosidase, and agarase that produced from Pseudomonas vesicularis MA103.  From the four hydrolysates, two enzymes were selected by higher reducing sugar contents, and the polysaccharide extracts of G. filicina were digested by the two enzymes.  The antioxidation of lactic acid fermented the hydrolysates was studied.  The polysaccharide extracts of G. filicina were digested by the same activity unit (100 unit) of those four enzymes.  The highest reducing sugar contents of those hydrolysates were digested by cellulase and agarase.  The larger the activity units of enzyme were, the higher the reducing sugars contents were.
| Six lactic acid (LA) fermentation products were cultured in media (1) the hydrolysate of 0.5% aqueous solution of polysaccharide extracts of G. filicina digested by 500 unit agarase and cellulase or (2) this G. filicina hydrolysate added 0.5% sucrose, 1% beef extract, and 0.5% yeast extract.(CN) And these two media was each fermented by three groups lactic acid bacteria, group A: 5% Lb. rhamnosus BCRC14068, group B: 5% Ent. faecalis BCRC13076, group C: 2.5% Lb. rhamnosus BCRC14068 and 2.5% Ent. faecalis BCRC13076.  The results of pH value of six LA fermentation products were decreasing as the time of fermentation increased.  The pH value of LA fermentation products that cultured in the hydrolysate added carbon and nitrogen source (L-Gra-CN-FC5 ) achieved below 4.6 at fermentation time of 9 hr.  That can be a secure product, and the titratable acidity is 0.74%, lactic acid bacteria count is 9.39 log CFU/mL
| Antioxidative properties, including ferrous ion chelating effect, inhibition of the hemoglobin-catalyzed peroxidation of linoleic acid, and reducing power of six LA fermentation products were better as the hydrolysate of G. filicina added carbon and nitrogen source than the hydrolysate of G. filicina, except for DPPH radical scavenging effect.  The soluble total polyphenol contents of the hydrolysate of G. filicina added carbon and nitrogen source that fermented by Lb. rhamnosus BCRC14068 and Ent. faecalis BCRC13076 was recorded 1,087.64 ± 0.31 GAE (gallic acid equivalents, μg/mL) higher than other fermentation products.  And the soluble total polyphenol contents of the other fermentation products were recorded from 400.27 ± 1.75 - 975.92 ± 4.90 GAE.  The antioxidative constituents of LA fermented G. filicina may be peptides, organic acid, and soluble total polyphenol contents.
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論文目次: | 目 錄
|
| 目錄    vi
| 表目錄    x
| 圖目錄    xii
| 一、前言    1
| 二、文獻整理    2
| I. 天然海藻類    2
| II. 蜈蚣藻簡介    2
| II-1. 蜈蚣藻型態與分布    2
| II-2. 蜈蚣藻成分    3
| II-3. 蜈蚣藻利用    4
| II-4. 蜈蚣藻之抗氧化特性    4
| (1) 自由基與活性氧之形成    4
| (2) 氧化傷害    5
| (3) 抗氧化劑的種類    6
| a. 自由基終止型    6
| b. 還原劑或氧清除劑    6
| c. 單重態氧抑制劑    6
| d. 金屬螯合劑    7
| e. 抗氧化酵素    7
| (4) 抗氧化活性測試原理    8
| a. α, α-Diphenyl-β-picrylhydrazl (DPPH) 自由基清除力指標及其原理    8
| b. 亞鐵離子螯合能力測試指標及其原理    8
| c. 還原力測試指標及其原理    9
| d. 抑制血紅素催化亞麻油酸自氧化測定指標及其原理    9
| (5) 蜈蚣藻的抗氧化成分    9
| II-5. 蜈蚣藻多醣體的結構與酵素的選用依據    10
| III. 乳酸菌簡介    10
| III-1. 乳酸菌的定義    10
| III-2. 乳酸菌之發酵特性    11
| III-3. 乳酸菌之抗氧化    12
| III-4. 乳酸菌的應用與生理功能    13
| 三、實驗設計    14
| 四、材料與方法    15
| I. 實驗材料    15
| I-1. 蜈蚣藻原料    15
| I-2. 試驗菌株及細胞株    15
| I-3. 培養基及藥品    15
| I-4. 儀器設備    19
| II. 實驗方法    19
| II-1. 蜈蚣藻儲存    19
| II-2. 樣品之一般成分分析    20
| (1) 水分含量    20
| (2) 灰份含量    20
| (3) 粗脂肪量    20
| (4) 粗蛋白含量    21
| (5) 醣量    21
| II-3. 乳酸菌之保存及活化    21
| (1) 乳酸菌之保存    21
| (2) 乳酸菌之活化    22
| II-4. 蜈蚣藻發酵液之製備及分析    22
| (1) 乳酸發酵前最適酵素水解物之製備    21
| (2) 蜈蚣藻多醣水溶液及其水解液經膠體過濾層析分析之測定    ...23
| (3) 薄層色層分析    24
| (4) 蜈蚣藻發酵議之製備    24
| (5) 乳酸菌菌數之測定    25
| (6)  pH 值之測定    25
| (7) 可滴定酸度之測定    25
| (8) 還原醣    26
| II-5. 蜈蚣藻乳酸發酵液之抗氧化活性    26
| (1) 清除α, α-Diphenyl-β-picrylhydrazl (DPPH) 自由
| 基能力之測定    26
| (2) 亞鐵離子螯合能力之測定    26
| (3) 還原力之測定    27
| (4) 亞麻油酸自氧化抑制力之測定    27
| (5) 總酚量之測定    28
| VI. 統計分析    28
| 五、結果與討論    29
| I. 蜈蚣藻冷凍乾燥粉末之一般成分分析    29
| II. 以四種多醣分解酵素降解蜈蚣藻多醣水溶液    29
| II-1. 還原醣含量變化    29
| II-2. 膠體過濾層析分析    30
| II-3. 薄層色層分析    33
| III. 經多醣分解酵素分解之蜈蚣藻多醣水溶液發酵實驗結果    33
| III-1. 乳酸菌接菌量之選用    34
| III-2.  pH值    35
| III-3. 可滴定酸度    35
| III-4. 乳酸菌數    36
| IV. 蜈蚣藻乳酸發酵液抗氧化性之探討    37
| IV-1. 清除 DPPH 自由基能力    38
| IV-2. 螯合亞鐵離子能力    38
| IV-3. 抑制血紅素催化亞麻油酸自氧化能力    39
| IV-4. 還原力    40
| V. 蜈蚣藻乳酸發酵液抗氧化性的成分    41
| 六、結論    43
| 七、參考文獻    44
|
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2010 年 3 月 8 日 23:00