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研究生:方翔陞
研究生(外文):FANG, XIANG-SHENG
論文名稱:金柑萃取物作為益生型乳酸菌的生長促進因子及抗氧化傷害保護劑之研究
論文名稱(外文):Studies on the Kumquat Extract as Protecting Agent for Oxidative Stress and Facilitating Factor for Lactic Acid Bacteria Growth
指導教授:林世斌林世斌引用關係陳莉臻陳莉臻引用關係
指導教授(外文):LIN, SHIH-BINCHEN, LI-CHEN
口試委員:孫舜國莊路德
口試委員(外文):SUN, SHUN-KUOCHUANGH, LU-TE
口試日期:2020-11-20
學位類別:碩士
校院名稱:國立宜蘭大學
系所名稱:食品科學系碩士班
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2021
畢業學年度:109
語文別:中文
論文頁數:73
中文關鍵詞:益生質乳酸菌抗氧化多酚金柑
外文關鍵詞:prebioticlactic acid bacteriaantioxidantpolyphenolkumquat
相關次數:
  • 被引用被引用:1
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  • 下載下載:64
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腸道菌群在人體腸道健康扮演不可或缺的角色,除了協助人體代謝之外,亦會影響免疫系統及神經系統。乳酸菌 (lactic acid bacteria, LAB) 大多屬於益生菌 (probiotic),為一類對人體腸道健康具有益處的菌群。由於人體在代謝過程會產生自由基,而過量的自由基將會引發人體內的氧化壓力,使腸道細胞及腸道菌群受到影響。已知多酚化合物 (polyphenol compounds, PCs) 具備良好的抗氧化活性,且 PCs 能夠改變腸道菌群的組成,並賦予宿主健康益處,亦屬於益生質 (prebiotic) 的範疇。由於金柑 (kumquat) 含有多種 PCs,本研究利用 Lactobacillus acidophilus、Lac. casei、Lac. helveticus、Lactobacillus plantarum 及 Lac. rhamnosus 等共六株乳酸桿菌,觀察金柑水萃取物 (kumquat extract by water, KE-W) 對乳酸桿菌生長的促進效果。結果顯示,KE-W 可促進所有試驗乳酸桿菌的生長,且有縮短菌株遲滯期 (lag phase) 之效果。利用極性差異經 C18 管柱劃分 KE-W 成金柑親水性劃分物 (KE-W/hydrophilicity, KE-W/HL) 及金柑疏水性劃分物 (KE-W/hydrophobicity, KE-W/HB),再經過生長促進的試驗結果發現,KE-W/HL 同樣具有促進試驗菌生長以及縮短遲滯期的能力。透過氧自由基吸收能力 (oxygen radical absorbance capacity, ORAC) 和鐵離子還原力 (ferric reducing antioxidant power, FRAP) 試驗探討 KE-W 及其劃分物的抗氧化力,得知三種樣品的抗氧化作用均為 KE-W/HB > KE-W > KE-W/HL,且樣品間具有顯著差異。由 100 mM AAPH 氧化傷害試驗可以發現,添加此三種樣品對於試驗菌株生長的保護作用可分成三個類型:第一類僅有 LA 10695 菌株,此三種樣品均有減緩其氧化傷害的程度;第二類包含 LH 14026、LP 10069 以及 LR 14029 等菌株,添加 KE-W 和 KE-W/HL 可降低 AAPH 造成的負面效果,但 KE-W/HB 則未有此作用;第三類則為 LC 10697 和 LR 16000,添加三種樣品均無法恢復 AAPH 對菌株造成之傷害。綜上所述,添加 KE-W 當中的 KE-W/HL 除了可以促進乳酸桿菌的生長之外,亦對氧化傷害的緩解有較佳的效果,可透過益生菌、益生質以及 PCs 的 3P 概念實現腸道健康的目標,對人體健康有產生益處之潛力。
Gut microbiota plays an indispensable role in our health. It can help the host with metabolic reactions. Moreover, it also affects the immune system and nervous system. Lactic acid bacteria (LAB) are a major group of probiotic which benefits to gut health. Metabolic reactions can generate free radical, which would cause oxidative stress and inhibit the growth of gut cell and microbiota. Polyphenol compounds (PCs) are known as strong antioxidants and can effectively scavenge free radical. Besides, PCs are also known as prebiotics and can stimulate gut bacteria and has beneficial to healthy. Kumquat, a locally characteristic, has been well known fruit containing multiple PCs.This study explored the potential growth facilitating effect of kumquat water extract (KE-W) to Lactobacillus acidophilus, Lac. casei, Lac. helveticus, Lac. plantarum, and Lac. rhamnosus. The results demonstrated that the bacteria number was increased and lag phase duration were significantly shortened by adding KE-W. The KE-W was further fractionated with the C18 column into hydrophilic and hydrophobic fractions designed as KE-W/HL and KE-W/HB, respectively. Results revealed that KE-W/HL has the growth facilitating effect to LAB. The antioxidant capability of KE-W, KE-W/HL and KE-W/HB were analyzed with oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) methods:the ORAC and FRAP value demonstrated their antioxidant capability as KE-W/HB > KE-W > KE-W/HL. The protection of these two fractions to the tested bacteria by under the oxidative stress caused by AAPH was also explored. The results showed that these LAB can be divided into three types against 100mM AAPH. The type 1:LA 10695 only, which can be protected with these three kinds of kumquat samples. LH 14026, LP10069 and LR 14029 are belong to type 2 that can mitigate the oxidative stress by KE-W and KE-W/HL instead of KE-W/HB. The type 3:LC 10697 and LR 16000, which can not be recovered by all the samples. In conclusion, KE-W/HL potentially have the capability to promote the growth and/or protect LAB under the oxidative stress.
摘要 I
Abstract II
誌謝詞 III
目次 IV
表次 VII
圖次 VIII
附圖目錄 IX
縮寫表 X
壹、前言 1
貳、文獻回顧3
一、腸道菌群及腸道環境對人體健康的重要性3
(一)腸道菌群對人體健康的影響3
(二)腸道環境與人體健康之關係5
二、益生菌及益生質與腸道健康之關聯6
三、乳酸菌對腸道健康的影響8
(一)乳酸菌概述8
(二)乳酸桿菌屬 (Lactobacillus) 的功能與應用性9
四、柑橘類水果多醣成分及多酚化合物之功效 12
(一)多醣成分 12
(二)多酚化合物 13
參、實驗架構 15
肆、材料與方法 16
一、 實驗材料 16
(一) 金柑 16
(二) 試驗菌株 16
二、 儀器設備 16
三、 藥品試劑及緩衝液之配製方法17
(一) 藥品試劑 17
(二) 緩衝液之配製方法 18
四、 微生物培養液及其配製方法 19
五、 實驗方法 19
(一) 金柑萃取物之製備 19
(二) 金柑萃取物多酚化合物之高效液相層析法分析20
(三) 總多酚含量測定 21
(四) 抗氧化能力分析 21
(五) 金柑水萃取物親水性及疏水性成分之劃分 22
(六) 金柑水萃取物對乳酸菌生長之影響 23
(七) 統計分析 26
伍、結果與討論 27
一、 兩種不同萃取溶劑對金柑總多酚含量及種類之影響27
(一) 總多酚含量 27
(二) 多酚化合物主成分分析 28
二、 金柑水萃取物 (KE-W) 對乳酸菌生長之促進效果 29
三、 KE-W 劃分物之總多酚含量、成分種類以及乳酸菌生長促進之比較31
(一) KE-W 劃分物總多酚含量與多酚化合物主成分分析31
(二) KE-W 劃分物對乳酸菌生長促進效果32
四、 KE-W 劃分物抗氧化能力及其氧化傷害保護之相關性33
(一) 劃分物之抗氧化能力34
(二) 劃分物對乳酸桿菌氧化傷害之保護效果35
陸、結論 38
柒、參考文獻39
捌、表63
玖、圖64
壹拾、附錄72

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