游若篍Yu, Roch-Chui臺灣大學:食品科技研究所孫豫蘋Sun, Yu-PingYu-PingSun2010-05-112018-06-292010-05-112018-06-292009U0001-1901200922044400http://ntur.lib.ntu.edu.tw//handle/246246/182197本研究利用乾鹽法（dry-salt method）以及簡單的配料來進行韓式口味之乳酸發酵白菜（lactic acid fermented cabbage）的製造，探討食鹽與溫度發酵對乳酸發酵白菜之影響，並探討發酵過程、添加雙叉桿菌對乳酸發酵白菜抗氧化活性以及乳酸發酵白菜於貯存過程中之菌相變化。酸發酵白菜隨著發酵時間增加，可滴定酸度增加、pH值下降；乳酸菌在發酵前期很快的成為主要優勢菌種，其中Leuconostoc mesenteroides為發酵初期的主要菌種，之後由Lactobacillus plantarum取而代之；發酵溫度對乳酸發酵白菜的乳酸菌相影響較食鹽明顯，溫度較高（25℃）時，L. mesenteroides與L. plantarum消長速度較快；發酵溫度較低（18℃）時，食鹽對菌數影響較顯著。乳酸發酵白菜經消費者喜好性評估，結果顯示以3%食鹽處理、25℃發酵溫度者較受消費者喜愛。氧化活性方面，萃取溶劑與發酵過程對白菜混合物（未發酵）的抗氧化活性有明顯的影響。ㄧ般而言，發酵過程對於白菜混合物之甲醇萃取物的螯合亞鐵離子能力與還原力並無影響，然而水萃取物部分之抗氧化活性卻有明顯的降低，甲醇萃取物比水萃取物具有較高的捕捉DPPH自由基能力與還原力, 其中發酵白菜的甲醇萃取物具有最高的捕捉DPPH自由基能力。萃取溶劑的種類與發酵過程對於萃取物中的總酚與類黃酮含量亦有影響，發酵過程會增加甲醇萃取物中的總酚含量，但卻會降低水萃取物中的類黃酮的含量，但大白菜混合物與乳酸發酵白菜萃取物的抗氧化變化與其中所含的總酚與類黃酮含量變化並不完全ㄧ致。加益生性雙叉桿菌Bifidobacterium breve BCRC 11846、B. lactis Bb-12、B. infantis BCRC 14602以及B. bifidum BCRC 14615至乳酸發酵白菜中，清除DPPH自由基清除能力情形，除了B. breve 11846與未添加之控制組的能力無顯著差異，其他添加雙叉桿菌之乳酸發酵白菜對於捕捉DPPH．能力較控制組具有顯著的促進效果；添加雙叉桿菌組的水萃取之螯合亞鐵離子能力明顯較未添加者佳，無論是甲醇萃取物或是水萃取物以添加B. infantis BCRC14602表現最佳，與其他組別具有顯著差異（p < 0.05）；還原力方面，添加雙叉桿菌之乳酸發酵白菜的水萃取物明顯較未添加之控制組的還原力好（p < 0.05），甲醇萃取物的還原力比水萃取物為佳，其中以添加B. lactis Bb12的乳酸發酵白菜甲醇萃取物表現最佳。綜合抗氧化實驗結果顯示添加雙叉桿菌的乳酸發酵白菜具有促進抗氧化能力效果，但因添加之雙叉桿菌菌種不同亦會有所差異。存過程對添加雙叉桿菌的乳酸發酵白菜的酸鹼值與可滴定酸方面，無顯著影響，但其中總乳酸菌數與雙叉桿菌的存活有明顯的影響，乳酸菌數由原先8 log CFU/g貯存至第三週時已減少至6~7 log CFU/g，有明顯差異（p < 0.05）；雙叉桿菌數方面，貯存至第2週時均減少至4 log CFU/g，有明顯的差異（p < 0.05）。抗氧化能力方面，無論有無添加雙叉桿菌之乳酸發酵白菜之甲醇萃取物的抗氧化能力（捕捉DPPH．能力與還原力），並未隨著貯存時間增加有明顯變化，但是水萃取物之抗氧化能力隨著貯存時間增加有明顯減少的變化。This study tries to produce lactic acid fermented cabbage similar to kimchi in Korea with a dry-salt method and simple formula, investigate the effects of temperature and NaCl on lactic acid fermented cabbage. Besides, antioxidant activity was determined and the effects of fermentation on the change of antioxidant activity, the antioxidative activity of supplemented with bifidobacteria into lactic acid fermented cabbage and storage were also investigated.esults showed that pH value decreased and titratable acidity increased following with fermentation time. Lactic acid bacteria (LAB) became the dominant flora after 1 day. Leuconostoc mesenteroides appeared at the earlier stage of fermentation and Lactobacillus plantarum appeared in the later stage. The changes of microflora during fermentation were much more dependent on temperature than on salt concentrations. L. mesenteroides decreased much more rapidly at 25℃ than at 18℃. The microbial growth in lactic acid fermented cabbage fermentation was significantly changed by sodium chloride within the range of 2-3% at 18℃. However, the influence of salt concentration was not significant while fermentation was carried out at 25℃. Hedonic sensory evaluation of lactic acid fermented cabbage was prepared at either 2% or 3% salt concentration and fermented at 25℃ for 2 day and 18℃ for 2.5 day. It showed that the lactic acid fermented cabbage dehydrated by 3% salt concentration, and fermented at 25℃ was better than others.ntioxidant activity observed on Chinese cabbage mixture may vary with extraction solvents and fermentation. Generally, the methanol extract of cabbage mixture showed a higher DPPH radical scavenging activity and reducing activity than did the water extract. Although fermentation did not alter the Fe2+-chelating ability and reducing activity of the methanol extract of the cabbage mixture, it reduced these same antioxidant activities in the water extract. Among the various extracts examined, the methanol extract of fermented cabbage showed the highest DPPH-radical scavenging effect. Additionally, the type of solvent and fermentation were also found to affect the total phenolic and flavonoid content of the extracts. Fermentation increased the total phenolic content of the methanol extract, while reducing the total flavonoid content of the water extract. Furthermore, changes in the antioxidant activity observed on the extracts of cabbage mixture and fermented cabbage did not coincide exactly with the content of total phenolics and total flavonoids.upplemented with B. breve BCRC 11846、B. lactis Bb-12、B. infantis BCRC 14602以及B. bifidum BCRC 14615 into lactic acid fermented cabbage to 107 CFU/g, there is no significantly difference on total lactic acid bacteria, L. mesenteroides, L. plantarum, pH value and titratable acid. The extracts of supplemented probiotic bifidobacteria of lactic acid fermented cabbage except supplemented B. breve 11846 showed a higher DPPH radicals scavenging effect than control (without supplemented). Moreover, the extracts of lactic acid fermented cabbage supplemented B. infantis BCRC14602 showed the best Fe2＋-chelating ability. The methanol extract of lactic acid fermented cabbage supplemented B. lactis Bb12 showed the best reducing power and the water extracts of lactic acid fermented cabbage supplemented bifidobacteria showed a higher DPPH radicals scavenging effect than control. Furthermore, supplemented bifidobacteria to lactic acid fermented cabbage may enhance the antioxidative activity, but have some variations in variety of bifidobacteria supplemented.hanges of lactic acid fermented cabbage supplemented with bifidobacteria on the total lactic acid bacteria, bifidobacteria, and antioxidative activity during storage at 4℃. there are no significant change at pH value and titratable acid, total lactic acid bacteria of fermented cabbage supplemented with/without bifidobacteria showed that there was not a significant decrease during the first 2 weeks while being storage at about 8 log CFU/g. However, there was a significant decrease to 6~7 log CFU/g at the third week (p<0.05), and an even more significant decrease to the remaining 4~5 log CFU/g at the fourth week (p<0.05). Form the second week the numbers of bifidobacteria were significantly decreased to 4 log CFU/g, but and antioxidative activity of methanol extracts showed no significant decrease or change during storage, but the water extracts significantly decreased during storage and the longer the storage time, the less antioxidative activity.論文口試委員審定書--------------------------------------------------------------------- i誌------------------------------------------------------------------------------------------ ii文摘要------------------------------------------------------------------------------------ iii文摘要------------------------------------------------------------------------------------ v錄------------------------------------------------------------------------------------------ vii次------------------------------------------------------------------------------------------ x次------------------------------------------------------------------------------------------ xi壹章、文獻整理------------------------------------------------------------------------ 1一、益生菌-------------------------------------------------------------------------------1. 益生菌的定義------------------------------------------------------------------------ 1. 乳酸菌之定義與特性--------------------------------------------------------------- 1 3. 乳酸菌之分類------------------------------------------------------------------------2 4. 乳酸菌之生理功效-----------------------------------------------------------------2 4.1 維持腸道中正常微生物相，促進腸胃道的健康------------------------- 3 4.2 改善乳糖不耐症、維生素的合成與改善營養價值--------------------- 3 4.3 調節免疫機能------------------------------------------------------------------ 3 4.4 降低血清中膽固醇含量------------------------------------------------------ 4 4.5 抗氧化--------------------------------------------------------------------------- 4 4.6 抗致突變與抗腫瘤------------------------------------------------------------ 5二、乳酸菌與乳酸發酵蔬菜---------------------------------------------------------- 6 1. 乳酸菌在食品中之作用----------------------------------------------------------- 6 2. 乳酸發酵蔬菜----------------------------------------------------------------------- 6 3. 影響蔬菜發酵之因子-------------------------------------------------------------- 7 3.1 食鹽濃度------------------------------------------------------------------------ 7 3.2 酸度------------------------------------------------------------------------------ 8 3.3 溫度------------------------------------------------------------------------------ 8 4. 韓式泡菜(Kimchi)------------------------------------------------------------------ 8 4.1 韓式泡菜製造過程------------------------------------------------------------ 9 4.2 韓式泡菜發酵過程------------------------------------------------------------ 11 4.3 韓式泡菜發酵的影響因子--------------------------------------------------- 11 4.4 營養與機能特性----------------------------------------------------------------- 12三、氧化作用及抗氧化機制原理---------------------------------------------------- 15 1. 自由基終止劑(Free radical terminator)-------------------------------------- 15 2. 還原劑或氧清除劑(reductants and oxygen scavengers)------------------- 15 3. 金屬螯合劑(metal chelator)---------------------------------------------------- 16四、蔬果中的抗氧化物質------------------------------------------------------------- 16 1. 維生素C (ascorbic acid, 抗壞血酸)----------------------------------------- 16 2. 維生素 E(生育醇, tocopherol)------------------------------------------------ 17 3. 類胡蘿蔔素(carotenoids)------------------------------------------------------- 17 4. 酚類化合物(phenolic compounds)-------------------------------------------- 17五、參考文獻---------------------------------------------------------------------------- 19貳章、溫度、食鹽對乳酸發酵大白菜之影響------------------------------------ 28Abstract ----------------------------------------------------------------------------------- 29摘要 --------------------------------------------------------------------------------------- 30前言---------------------------------------------------------------------------------------- 31材料與方法------------------------------------------------------------------------------- 33結果與討論------------------------------------------------------------------------------- 37結論 --------------------------------------------------------------------------------------- 48參考文獻---------------------------------------------------------------------------------- 49參章、乳酸發酵大白菜的抗氧化性研究------------------------------------------ 52Abstract ----------------------------------------------------------------------------------- 53摘要 --------------------------------------------------------------------------------------- 54前言---------------------------------------------------------------------------------------- 55材料與方法------------------------------------------------------------------------------- 57結果與討論------------------------------------------------------------------------------- 60結論---------------------------------------------------------------------------------------- 71參考文獻---------------------------------------------------------------------------------- 72肆章、添加雙叉桿菌之乳酸發酵大白菜的抗氧化特性探討----------------- 77Abstract ----------------------------------------------------------------------------------- 78摘要 --------------------------------------------------------------------------------------- 79前言---------------------------------------------------------------------------------------- 80材料與方法------------------------------------------------------------------------------- 82結果與討論------------------------------------------------------------------------------- 85結論 -------------------------------------------------------------------------------------- 102參考文獻---------------------------------------------------------------------------------- 103伍章、添加雙叉桿菌之乳酸發酵大白菜於4℃貯存期間之抗氧化變化--- 107Abstract ---------------------------------------------------------------------------------- 108摘要 -------------------------------------------------------------------------------------- 109前言--------------------------------------------------------------------------------------- 110材料與方法------------------------------------------------------------------------------- 111結果與討論------------------------------------------------------------------------------- 114結論 -------------------------------------------------------------------------------------- 124參考文獻---------------------------------------------------------------------------------- 125錄1、溫度與食鹽對乳酸發酵白菜之發酵過程的菌相變化------------------ 128錄2、乳酸發酵大白菜之消費者喜好性品評問卷------------------------------ 129錄3、Antioxidant activity of lactic-fermented Chinese cabbage(已接受)----- 131次1.1 韓式泡菜之製作流程----------------------------------------------------------- 102.1 食鹽、發酵溫度對發酵大白菜之發酵期間的pH 值之變化----------- 382.2 食鹽、發酵溫度對發酵大白菜之發酵期間的可滴定酸度之變化------ 402.3 不同食鹽量預處理(2%, 3%)、溫度對乳酸發酵大白菜發酵過程 菌相變化之影響----------------------------------------------------------------- 443.1 大白菜混合物與發酵大白菜之甲醇萃取物與水萃取物之 DPPH自由基清除能力--------------------------------------------------------- 653.2 大白菜混合物與發酵大白菜之甲醇萃取物與水萃取物之亞 鐵離子螯合能力----------------------------------------------------------------- 663.3 大白菜混合物與發酵大白菜之甲醇萃取物與水萃取物之還 原力--------------------------------------------------------------------------------- 674.1 添加雙叉桿菌的發酵大白菜甲醇萃取物之DPPH自由基清 除能力----------------------------------------------------------------------------- 914.2 添加雙叉桿菌的發酵大白菜水萃取物之DPPH自由基清除 能力-------------------------------------------------------------------------------- 924.3 添加雙叉桿菌的發酵大白菜甲醇萃取物之亞鐵離子螯合能力-------- 954.4 添加雙叉桿菌的發酵大白菜水萃取物之亞鐵離子螯合能力----------- 964.5 添加雙叉桿菌的發酵大白菜甲醇萃取物之還原力----------------------- 984.6 添加雙叉桿菌的發酵大白菜水萃取物之還原力-------------------------- 995.1 添加雙叉桿菌之發酵大白菜於4℃貯存過程中的酸鹼值與可 滴定酸之變化-------------------------------------------------------------------- 1175.2 添加雙叉桿菌的發酵大白菜於貯存期間甲醇萃取物之DPPH 自由基清除能力的變化------------------------------------------------------- 1205.3 添加雙叉桿菌的發酵大白菜於貯存期間水萃取物之DPPH自 由基清除能力的變化---------------------------------------------------------- 1215. 4 添加雙叉桿菌的發酵大白菜於貯存期間甲醇萃取物之還原能 力的變化------------------------------------------------------------------------ 1225.5 添加雙叉桿菌的發酵大白菜於貯存期間水萃取物之還原能力 之變化------------------------------------------------------------------------------ 1231.1 韓式泡菜中的生理活性物質------------------------------------------------- 142.1 喜好性品評之發酵大白菜之酸鹼值與可滴定酸度情形---------------- 462.2 2%、3%鹽處理與25℃、18℃發酵溫度製備之發酵大白菜 的喜好性品評------------------------------------------------------------------- 473.1 大白菜混合物與發酵大白菜之萃取率------------------------------------- 613.2 大白菜混合物與發酵大白菜之甲醇萃取物與水萃取物之抗 氧化特性------------------------------------------------------------------------- 683.3 大白菜混合物與發酵大白菜之甲醇萃取物與水萃取物之總 酚與類黃酮含量情形---------------------------------------------------------- 704.1 添加雙叉桿菌之發酵大白菜的總乳酸菌菌數、雙叉桿菌數 、L. mesenteroides 與 L. plantarum菌數情形---------------------------- 874.2 添加雙叉桿菌之發酵大白菜的酸鹼值與可滴定酸度情形-------------- 884.3 添加雙叉桿菌的發酵大白菜甲醇萃取物與水萃取物對捕 捉DPPH．能力與螯合亞鐵離子能力之半有效濃度(IC50)------------ 1015.1 添加雙叉桿菌之發酵大白菜於4℃貯存期間之的總乳酸菌 菌數、雙叉桿菌數變化情形------------------------------------------------- 116application/pdf1489792 bytesapplication/pdfen-US乳酸發酵大白菜乾鹽法捕捉DPPH自由基能力還原力螯合亞鐵離子能力lactic-fermented Chinese cabbagedry-salt methodDPPH-radical scavenging effectFe2+-ion chelating abilityreducing activitiesthesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/182197/1/ntu-98-D89641006-1.pdf