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第 28 筆
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論文名稱: | 乳酸菌發酵糙薏仁優格之產製技術及生理活性探討
| Studies on the Production Technology and Biological Activities of Lactic Acid Bacteria Fermented Dehulled Adlay Yogurt
研究生: | 黃祥凱 Siang-Kai Huang
指導教授: | 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: | M91320055
畢業學年度: | 92
語文別: | 中文
關鍵字: | 糙薏仁 dehulled adlay
| 優格 yogurt
| 乳酸菌 lactic acid bacteria
全文點閱次數: | 15

全文影像連結:

論文全文(pdf)

論文頁數: | 165
摘要: | 摘要
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| 本研究分別以台中一號 (Tc1)、台中選育四號 (Ts4)、泰國進口 (Th)、及寮國進口 (La) 四種糙薏仁優格接種 2% 乳酸菌菌量進行發酵。經三組乳酸菌菌組發酵後，發現添加 3 g、4 g、及 5 g 四種糙薏仁優格以 Lactobacillus (Lb.) plantarum BCRC10069 (1%) 和 BCRC12250 (1%) 於 37oC 發酵 24 hr後，pH 值可降至 4.6 以下，可滴定酸度為 0.78-0.87%，而乳酸菌菌數增殖至 8 log CFU/g 以上。3-5 g 四種糙薏仁優格冷水萃取物 (12.5 mg/mL) 進行三種抗氧化試驗，在清除 DPPH 自由基能力中以 5 g 台中一號糙薏仁優格 (Tc1-f) 效果較佳為 10.20%；在螯合亞鐵離子能力方面，四種糙薏仁優格螯合能力皆可達 60% 以上；而在抑制血紅素催化亞麻油酸自氧化能力方面，四種糙薏仁優格呈現 11.96-104.81% 之抑制能力。四種糙薏仁優格於 4oC 貯藏 14 天過程中，pH 值自原先 4.54-4.63 降至 4.32-4.42，可滴定酸度由 0.77-0.78% 增加至 0.95-1.02%，而乳酸菌菌數仍可維持 8 log CFU/g 以上，至於清除 DPPH 自由基與螯合亞鐵離子能力則會隨貯藏時間增加而逐漸減少。安氏試驗法以 S. typhimurium TA100 為測試菌株，結果發現 3-5 g 四種糙薏仁優格冷水萃取物對致突變劑 4NQO 及 B[a]P 皆有抗致突變效果，分別為 24.0-32.0% 與 24.2-42.1%。動物細胞生理活性試驗中，泰國 (Th-f) 與寮國糙薏仁優格 (La-f) 對人類融合瘤細胞株 HB4C5 具有增生效果，而在人類單核球細胞株 THP-1 方面，四種糙薏仁優格皆具增生能力。SD 大白鼠動物實驗發現，餵食糙薏仁優格組之大白鼠肝臟重及體重皆較控制組為低，且餵食 Th-f 會降低大白鼠肝臟總膽固醇與三酸甘油酯含量。在肝功能指標酵素 AST 及 ALT 方面，糙薏仁優格的飲食會降低其濃度，並增加大白鼠糞便排出量。在糞便總膽固醇及三酸甘油酯含量以餵食臺中選育四號糙薏仁優格 (Ts4-f) 為最低。至於在腸道生理方面，二組糙薏仁優格能明顯增加大白鼠盲腸內容物含量、降低盲腸 pH 值，在盲腸相關菌數中發現餵食糙薏仁優格可增加大白鼠盲腸 Lactobacillus (7.37-7.72 log CFU/g)、Bifidobacterium (7.41-8.78 log CFU/g)、及 total anaerobes (8.72-9.85 log CFU/g) 菌數，並減少 C. perfrigens (5.39-3.50 log CFU/g) 菌數。
|
| Abstract
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| Lactic acid bacteria (LAB) were used to ferment four dehulled adlay yogurts. Four dehulled adlay are: Taichung No. 1 (Tc1), Taichung Shuenyu No. 4 (Ts4), Thailand imported (Th), and Laos imported (La).  The 3 g, 4 g, and 5 g four dehulled adlay yogurts that pH value under 4.6, titrable acid for 0.78-0.87%, and LAB counts above 8 log CFU/g, by Lactobacillus (Lb.) plantarum BCRC10069 (1%) and BCRC12250 (1%) fermented at 37oC for 24 hr.  Four dehulled adlay yogurt cold water extracts (12.5 mg/mL) were proceeded three antioxidant test.  The ability for scavenging DPPH free radical of 5 g Taichung No. 1 dehulled adlay yogurt (Tc1-f) was better, as 10.20%.  As to the ability of chelating on Fe (II) ion, the results indicated that four dehulled adlay yogurts performed above 60% chelating effect on Fe (II) ion.  In the antioxidant effect as measurement on inhibition of hemoglobin-induced linoleic acid oxidation, the results showed four dehulled adlay yogurts as 11.96-104.81% antioxidant activity.  Four dehulled adlay yogurts that pH value from original 4.54-4.63 falls to 4.32-4.42, titrable acidity from 0.77-0.78% increases to 0.95-1.02%, LAB counts still maintain above 8 log CFU/g during 4oC for 14 days.  The ability of scavenging DPPH free radical and chelating on Fe (II) ion with store time increase, the antioxidant activity was gradually decreased.  S. typhimurium TA100 was used in the Ames test, and the results showed 3-5 g four dehulled adlay yogurt cold water extracts were antimutagenicity ability to 4NQO and B[a]P.  In the immuno-cells physiology activity experiment, both Thailand (Th-f) and Laos’s dehulled adlay yogurts (La-f) were growth-promotion ability to human hybridoma HB4C5 cell, and to the human acute monocyte THP-1 cell, four dehulled adlay yogurts were growth-promotion ability.  SD rats experience result discovers that yogurt diet had lower relative hepatic weight and body weight when compared with those animals fed with the control diet.  Th-f diet had lower in liver cholesterol and triglyceride content.  Dehulled adlay yogurt diet induced a decrease liver AST and ALT levels, and increase fecal content of rats. In addition, rats fed the Taichung Shuenyu No. 4 dehulled adlay yogurt (Ts4-f) diet had higher in fecal cholesterol and triglyceride content.  A significant increase in cecal contents and a significant decrease in fecal pH value were observed in rats fed with the yogurt diet.  It was interesting to note that rats fed with the dehulled adlay yogurt diet had elevated microbial count on fecal Lactobacillus (increased from 7.37 to 7.72 log CFU/g), Bifidobacterium (increased from 7.41 to 8.78 log CFU/g), and total anaerobes (increased from 8.72 to 9.85 log CFU/g), as depressed Clostridium (C.) perfringens count (decreased from 5.39 to 3.50 log CFU/g).
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論文目次: | 目 錄
| 目    錄    i
| 表 目 錄    viii
| 圖 目 錄    x
| 中文摘要    xi
| 英文摘要    xii
| 一、前言    1
| 二、文獻整理    3
| I.    薏苡之生理功能    3
| I-1.    薏苡簡介    3
| I-2.    薏苡之本草綱要    3
| I-3.    薏苡之成分    4
| II.    薏苡之生理機能性    6
| II-1.    抗氧化作用    6
| II-2.    抗致突變作用    7
| II-3.    免疫調節作用    7
| II-4.    抗癌作用    8
| II-5.    調節血脂    9
| II-6.    調節血糖    10
| II-7.    對腸道生理的影響    11
| III. 乳酸菌之特性、利用及功能性    11
| III-1. 乳酸菌之定義    11
| III-2. 乳酸菌之發酵特性    12
| III-3. 乳酸菌之生理功能    12
| III-3-1.    兼性厭氧性與芳香物質之生成    12
| III-3-2.    提高營養價值、促進維生素的合成及
| 酵素的產生    13
| III-3-3.    具抗菌作用    13
| III-3-4.    具保健特性    13
| IV.    乳酸菌所產胞外多醣對優格質地之影響    14
| IV-1.    Ropy 及 non-ropy 乳酸菌對優格微細結構之影響    14
| IV-2.    乳酸菌所產不同型態 EPS 對優格質地改變之影響    15
| IV-3.    乳酸菌所產 EPS 對優格品質之影響    16
| V.    乳酸菌之抗氧化    17
| VI.    乳酸菌及其發酵乳製品之抗致突變性研究    18
| VI-1.    抗致突變物之作用機制    19
| VI-2.    去致突變物    20
| VI-2-1.    防止致突變物生成之抑制劑    20
| VI-2-2.    降低致突變物造成 DNA 傷害之阻斷劑    20
| VI-2-2-1.    致原致突變物之不活化作用    20
| VI-2-2-2.    對代謝活化酵素系統之抑制作用    20
| VI-2-2-3.    對原致突變物代謝活化物之
| 不活化作用    20
| VI-2-2-4.    促進解毒代謝系統    21
| VI-2-3.    抗氧化劑    21
| VI-3.    生物抗致突變物    21
| VI-3-1.    作用於 DNA 修補之化合物    21
| VI-3-2.    抑制腫瘤促進與增殖作用之抑制劑    22
| VII.    抗致突變性之檢測方法    22
| VII-1.    Histidine requirement    23
| VII-2.    rfa mutation    23
| VII-3.    uvrB mutation    23
| VII-4.    R-factor    23
| VII-5.    pAQ1 plasmid    24
| VIII. 乳酸菌對 pH 值、膽鹽與肝臟脂質之影響    24
| VIII-1.    pH 值、膽鹽    24
| VIII-2.    肝臟脂質    25
| VIII-3.    血脂質    25
| VIII-4.    糞便脂質    26
| IX.    乳酸菌對腸道生理的影響    26
| IX-1.    附著腸道的能力    26
| IX-2.    腸道菌相    27
| IX-3.    盲腸與糞便 pH 值    27
| X.    免疫活性物質之評估方法    28
| X-1.    免疫細胞之活化    28
| X-1-1.    T 細胞之活化    28
| X-1-2.    自然殺手細胞之活化    28
| X-1-3.    吞噬細胞之活化    29
| X-2.    細胞存活率 (MTT 還原分析法)    29
| 三、實驗設計    30
| 四、材料與方法    32
| I.    實驗材料    32
| I-1.    糙薏仁粉末    32
| I-2.    試驗菌株    32
| I-3.    培養基    32
| I-4.    藥品    35
| I-5.    材料與儀器設備    39
| II.    乳酸菌菌株之保存與活化    40
| II-1.    菌株的保存    40
| II-2.    菌株的活化    40
| III.    乳酸菌發酵糙薏仁優格之最適條件探討    41
| III-1.    糙薏仁優格之製備    41
| III-2. 微生物含量之測定    41
| III-3.    糙薏仁優格可滴定酸度之測定    41
| III-4.    糙薏仁優格之貯藏試驗    42
| VI.    糙薏仁粉末及糙薏仁優格凍乾粉末之一般成分分析    42
| VI-1.    水分含量    42
| VI-2.    灰分含量    42
| VI-3.    粗脂肪含量    43
| VI-4.    粗蛋白質含量    43
| VI-5.    無氮萃取物含量    43
| V.    糙薏仁粉末及糙薏仁優格之抗氧化性之探討    44
| V-1.    糙薏仁粉末及糙薏仁優格冷水萃取物之製備    44
| V-2.    清除 □,□-diphenyl-□-picrylhydrazyl (DPPH) 自由基
| 能力測定    44
| V-3.    抑制血紅素催化亞麻油酸自氧化測定    44
| V-4.    螯合亞鐵離子能力之測定    45
| VI.    安氏試驗法    45
| VI-1.    S. typhimurium TA100 之保存與活化    45
| VI-1-1.    試驗菌株之保存    45
| VI-1-2.    試驗菌株之活化    45
| VI-2.    S. typhimurium TA100 基因型態之確認    46
| VI-2-1.    組胺酸需求性 (histidine requirement) 之確認    46
| VI-2-2.    rfa 突變之測試    46
| VI-2-3.    uvrB 突變之測試    46
| VI-2-4.    R-factor 之測試    46
| VI-3.    毒性試驗    47
| VI-4.    致突變實驗    47
| VI-5.    抗致突變性試驗    48
| VII.    動物細胞生理活性測試    48
| VII-1.    培養動物細胞株    48
| VII-2.    培養基使用條件    49
| VII-3.    細胞存活率 (MTT assay)    49
| VIII.    SD 大白鼠生理活性測試    49
| VIII-1.    實驗動物    49
| VIII-2.    動物分組    49
| VIII-3.    動物飼養及樣品收集    50
| VIII-4.    血漿分析    50
| VIII-4-1.    總膽固醇 (total cholesterol) 之測定    50
| VIII-4-2.    三酸甘油酯 (triglyceride) 之測定    51
| VIII-4-3.    Aspartate transaminase (AST) 之測定    51
| VIII-4-4.    Alanine transaminase (ALT) 之測定    51
| VIII-4-5.    Thiobarbituric acid reactive substance (TBARS)
| 值之測定    52
| VIII-5.    肝臟之分析    52
| VIII-5-1.    肝臟脂質測定    52
| VIII-5-2.    Thiobarbituric acid reactive substance (TBARS)
| 值之測定    52
| VIII-6.    盲腸分析    53
| VIII-6-1.    盲腸 pH 值的測定    53
| VIII-6-2.    盲腸菌叢分析    53
| VIII-6-3.    選擇性培養基    53
| VIII-7.    糞便之分析    55
| VIII-7-1.    糞便脂質    55
| VIII-7-2.    糞便pH的測定    55
| VIII-7-3.    糞便濕重、乾重    55
| IX.    統計分析    55
| 五、結果與討論    56
| I.    糙薏仁粉末及糙薏仁優格凍乾粉末之一般成分    56
| II.    以三種乳酸菌之菌種組合發酵四種糙薏仁優格    56
| II-1.    乳酸菌菌株之選用    56
| II-2.    四種糙薏仁優格之發酵實驗結果    57
| III.    糙薏仁及糙薏仁優格抗氧化活性之探討    58
| III-1.    清除 DPPH 自由基之能力    58
| III-2.    亞鐵離子螯合效果    59
| III-3.    抑制血紅素催化亞麻油酸自氧化之能力    60
| VI.    糙薏仁優格貯藏試驗之探討    62
| VI-1.    貯藏期間 pH 值與可滴定酸度含量    62
| VI-2.    貯藏期間乳酸菌菌數與好氣性生菌數變化    62
| VI-3.    貯藏期間糙薏仁優格抗氧化能力之變化    63
| V.    糙薏仁及糙薏仁優格抗致突變性之探討    64
| V-1.    S. typhimurium TA100 試驗菌株基因型態之確認    64
| V-2.    毒性試驗    65
| V-3.    致突變性試驗    65
| V-4.    抗致突變性試驗    66
| VI.    四種糙薏仁及糙薏仁優格對動物細胞生理活性之影響    68
| VI-1.    對人類融合瘤細胞株 HB4C5 之細胞存活率影響    68
| VI-2.    對人類單核球細胞株 THP-1 之細胞存活率影響    68
| VII.    糙薏仁及糙薏仁優格餵食 SD 大白鼠四週後攝食量、
| 體重、及臟器重量之變化    70
| VII-1.    每日攝食量變化    70
| VII-2.    體重之變化    70
| VII-3.    肝臟之重量    71
| VIII.    餵食 SD 大白鼠糙薏仁及糙薏仁優格四週後肝臟脂質之
| 變化    71
| VIII-1.    膽固醇含量之變化    71
| VIII-2.    三酸甘油酯含量之變化    72
| VIII-3.    游離膽固醇、酯化型膽固醇含量與其比值之變化    72
| IX.    餵食 SD 大白鼠糙薏仁及糙薏仁優格四週後血漿中
| 葡萄糖含量及脂質之變化    74
| IX-1.    血漿中葡萄糖之含量    74
| IX-2.    血漿總膽固醇濃度之變化    74
| IX-3.    血漿三酸甘油酯濃度之變化    75
| X.    餵食 SD 大白鼠糙薏仁及糙薏仁優格四週後血漿中
| AST 及 ALT 活性之變化    75
| XI.    餵食 SD 大白鼠糙薏仁及糙薏仁優格四週後肝臟與
| 血漿 TBARS 之變化    76
| XI-1.    肝臟 TBARS 濃度之變化    76
| XI-2.    血漿 TBARS 濃度之變化    76
| XII.    餵食 SD 大白鼠糙薏仁及糙薏仁優格四週後糞便脂質之
| 變化    77
| XII-1.    糞便排出量之變化    77
| XII-2.    糞便 pH 值之變化    78
| XII-3.    糞便膽固醇含量    78
| XII-4.    糞便三酸甘油酯含量    78
| XII-5.    糞便膽酸含量    79
| XIII.    餵食 SD 大白鼠糙薏仁及糙薏仁優格四週後盲腸環境及
| 菌相之變化    79
| XIII-1.    盲腸重量之變化    79
| XIII-2.    盲腸 pH 值之變化    79
| XIII-3.    盲腸菌叢之變化    80
| XIII-3-1.    Clostridium prefrigens 菌數之變化    80
| XIII-3-2.    Lactobacillus 菌數之變化    80
| XIII-3-3.    Bifidobacterium 菌數之變化    80
| XIII-3-4.    Total anaerobes 菌數之變化    81
| 六、結論    82
| 七、參考文獻    84
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| 表 目 錄
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| 表一、        四種糙薏仁粉末及糙薏仁優格凍乾粉末之一般成分    104
| 表二、        以乳酸菌 Lb. bulgaricus BCRC10696 和 Str. thermophilus
| BCRC12268 發酵糙薏仁優格之 pH 值、可滴定酸度與
| 微生物菌數變化    ……………105
| 表三、        以乳酸菌 Lc. lactis BCRC12315 和 BCRC12322 發酵
| 糙薏仁優格之 pH 值、可滴定酸度與微生物菌數變化    106
| 表四、        以乳酸菌 Lb. plantarum BCRC10069 和 BCRC12250 發酵
| 糙薏仁優格之 pH 值、可滴定酸度與微生物菌數變化    107
| 表五、        四種糙薏仁冷水萃取物對 DPPH 自由基之清除效應    108
| 表六、        四種糙薏仁牛乳冷水萃取物對 DPPH 自由基之清除效應    109
| 表七、        四種糙薏仁優格冷水萃取物對 DPPH 自由基之清除效應    110
| 表八、        四種糙薏仁冷水萃取物對螯合亞鐵離子能力之測定    111
| 表九、        四種糙薏仁牛乳冷水萃取物對螯合亞鐵離子能力之測定    112
| 表十、        四種糙薏仁優格冷水萃取物對螯合亞鐵離子能力之測定    113
| 表十一、        四種糙薏仁冷水萃取物對抑制血紅素催化亞麻油酸
| 自氧化能力之影響    114
| 表十二、        四種糙薏仁優格冷水萃取物對抑制血紅素催化亞麻油酸
| 自氧化能力之影響    115
| 表十三、        四種糙薏仁冷水萃取物對 S. typhimurium TA100 之毒性    116
| 表十四、        四種糙薏仁優格冷水萃取物對 S. typhimurium TA100 之
| 毒性    117
| 表十五、        四種糙薏仁冷水萃取物對 S. typhimurium TA100 之
| 致突變性    118
| 表十六、        四種糙薏仁優格冷水萃取物對 S. typhimurium TA100 之
| 致突變性    119
| 表十七、        四種糙薏仁冷水萃取物對 S. typhimurium TA100 之
| 抗致突變性    120
| 表十八、        四種糙薏仁優格冷水萃取物對 S. typhimurium TA100 之
| 抗致突變性    121
| 表十九、        實驗飼料組成    122
| 表二十、        餵食 SD 大白鼠不同飼料四週後肝臟重量的變化    123
| 表二十一、SD 大白鼠餵食不同飼料四週後肝臟中膽固醇與
| 三酸甘油酯含量的變化    124
| 表二十二、SD 大白鼠餵食不同飼料四週後肝臟中游離膽固醇、
| 酯化型膽固醇濃度、與游離膽固醇比酯化型膽固醇
| 比值的變化    125
| 表二十三、餵食 SD 大白鼠不同飼料四週後血漿中總膽固醇及
| 三酸甘油酯的變化    126
| 表二十四、SD 大白鼠餵食不同實驗飼料四週後糞便 pH 值以及
| 一日糞便濕重與乾重的變化    127
| 表二十五、SD 大白鼠餵食不同飼料四週後糞便中膽固醇、
| 三酸甘油酯與膽酸濃度的變化    128
| 表二十六、SD 大白鼠餵食不同實驗飼料四週後盲腸 pH 值、
| 總重、內容物含量與盲腸壁重    129
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| 圖 目 錄
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| 圖一、    四種糙薏仁優格於 4oC 貯藏期間之 pH 值變化    130
| 圖二、    四種糙薏仁優格於 4oC 貯藏期間之可滴定酸度變化    131
| 圖三、    四種糙薏仁優格於 4oC 貯藏期間之乳酸菌菌數 (LAB)
| 變化    132
| 圖四、    四種糙薏仁優格於 4oC 貯藏期間之好氣性生菌數 (PCA)
| 變化    133
| 圖五、    四種糙薏仁優格於 4oC 貯藏期間清除 DPPH 自由基
| 能力之變化    134
| 圖六、    四種糙薏仁優格於 4oC 貯藏期間對亞鐵離子之螯合
| 能力變化    135
| 圖七、    四種糙薏仁對人類融合瘤細胞株 HB4C5 生長之影響    136
| 圖八、    四種糙薏仁優格對人類融合瘤細胞株 HB4C5 生長之影響    137
| 圖九、    四種糙薏仁對人類單核球細胞株 THP-1 生長之影響    138
| 圖十、    四種糙薏仁優格對人類單核球細胞株 THP-1 生長之影響    139
| 圖十一、    餵食 SD 大白鼠不同飼料之每日攝食量變化情形    140
| 圖十二、    餵食不同飼料四週期間 SD 大白鼠體重的變化情形    141
| 圖十三、    SD 大白鼠餵食不同實驗飼料四週後血漿中葡萄糖
| 含量的變化    142
| 圖十四、    餵食 SD 大白鼠不同飼料四週後血漿中天門冬胺酸
| 轉胺酶活性變化    143
| 圖十五、    餵食 SD 大白鼠不同飼料四週後血漿中丙胺酸轉胺酶
| 活性變化    144
| 圖十六、    SD 大白鼠餵食不同實驗飼料四週後肝臟中 TBARS
| 含量的變化    145
| 圖十七、    SD 大白鼠餵食不同實驗飼料四週後血漿中 TBARS
| 含量的變化    146
| 圖十八、    SD 大白鼠餵食不同飼料四週後盲腸部分菌群菌數的變化    147
|
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2010 年 3 月 8 日 23:02