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研究生:蔡淑雯
研究生(外文):Shu Wan Tsai
論文名稱:不同品種竹筍萃取液抗菌性之探討
論文名稱(外文):The Antibacterial Activities of Various Species of Bamboo Shoots Extracts
指導教授:謝寶全謝寶全引用關係
指導教授(外文):Pao Chuan Hsieh
學位類別:碩士
校院名稱:國立屏東科技大學
系所名稱:食品科學系
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:2002
畢業學年度:90
語文別:中文
論文頁數:191
中文關鍵詞:抗菌活性竹筍萃取液洋菜擴散法
外文關鍵詞:antibacterial activitybamboo shoot extractagar diffusion method.
相關次數:
  • 被引用被引用:9
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本研究係利用台灣省六種食用竹筍,分別為孟宗竹筍、烏腳綠竹筍、桂竹筍、麻竹筍、綠竹筍及箭竹筍等為試驗材料,所得之竹筍萃取物以洋菜擴散法(agar diffusion method)進行抗菌活性之測試,其試驗菌株以細菌九株為對象。其中以烏腳綠竹筍、麻竹筍及綠竹筍三種具較佳之抗菌活性,經50℃,24小時水萃取後可得最佳之萃取條件。進一步探討三種竹筍水萃取液在各種溫度、pH值、金屬離子及常用化學添加物對其抑菌活性之影響。其結果顯示,竹筍水萃取液經121℃,20分鐘加熱處理並不會破壞其抑菌成分;萃取液受pH值改變而顯著降低其抑菌活性;而添加金屬離子及化學添加物對竹筍萃取液會產生不同程度之抑菌活性影響。三種不同品種竹筍萃取液之最小抑制濃度MIC (mg/mL)分別為>500、155及195 mg/mL。於液體培養基中,分別添加2 %之三種竹萃取液及己二烯酸鉀對Listeria monocytogenes之抑制率分別為20.45、49.78、45.95及
16.62 %。竹筍水萃取液抑菌機制可能為細胞膜滲透壓改變,使細胞質流出,導致細胞死亡。而添加不同濃度之竹筍萃取液於4℃之市售奶茶中貯存5天,其生菌數皆低於控制組。
三種不同品種竹筍水萃取液以Sephadex G-25管柱層析分離純化抑菌成分,初步判定抗菌物質之分子量為528 Da。綠竹筍水萃取液以Amberlite XAD-7膠過濾層析後,分別可得區分A及區分E抑菌物質,再經Sephadex G-25、Sephadex G-15膠過濾層分離純化抑菌物質區分A-Ⅲ-3,經HPLC分析具3個吸收波峰。區分A-Ⅲ-3經紅外光光譜分析及氣相層析質譜儀分析,初步判定其抑菌物質為具有苯環之酚類物質。
The purpose of this reserch was to measure the antibacterial activities of six species of bamboo shoots (Phyllostachys edulis, Bambusa edulis, Phyllostachys bambusoides, Dendrocalamus latiflorus, Bambusa oldhamii and Fargesia spathaces). The antibacterial tests were carried out by using agar diffusion method. The extracts of Bambusa edulis, Dendrocalamus latiflours and Bambusa oldhamii had stronger antibacterial activities. The extracts of bamboo shoots were extracted with distilled water at 50℃ for 24 hours. The effects of temperatures, pH, metal ions and chemical additives were studied on antibacterial activities of bamboo shoot extracts. After extracts of bamboo shoots were treated with 121℃, 20min, the results revealed that extracts of bamboo shoots still possessed antibacterial activities. The antibacterial activities decreased obviously with pH values changed. Adding metal ions and chemical additives on extracts showed different antibacterial activities. The extracts of bamboo shoots had the MIC (mg/mL) >500, 155 and 195mg/mL respectively. The extracts of bamboo shoots and potassium sorbate were added in the broth of Listeria monocytogenes, the inhibition ratio were 20.45, 49.78, 45.59 and 16.62%, respectively. The mechanism of inhibition may be related to the leakage of the cell wall or probably some alteration of the membrane permeability and thereby resulting in the loss of the cytoplasm, and then causing the death of the cells. To study the preservative agent on bamboo shoots in milk tea added with bamboo shoot extracts at 4℃ for 5days, the total counts were lower than that of control.
Isolation and purification of antibacterial compounds were studied for three species of bamboo shoots by Sephadex G-25. The molecular weight of antibacterial compound was estimated to be 270Da. Amberlite XAD-7 column chromatography, Sephadex G-25, Sephadex G-15 chromatography were chosen for the extract of Bambusa oldhamii. Fraction A and fraction E from Amberlite XAD-7 chromatography were purified continuously by Sephadex G-25 and Sephadex G-15 column chromatography. Active fraction A-Ⅲ-3 were obtained and assayed by HPLC. A-Ⅲ-3 had three peaks. The antibacterial compound was identified by IR and GC-MS. The results showed that the antibacterial compound may was the phenolic compound and identified to contain benzene.
中文摘要-------------------------------------------------------------- I
英文摘要-------------------------------------------------------------- III
誌謝---------------------------------------------------------------------- Ⅴ
目錄-------------------------------------------------------------------- VI
表目錄--------------------------------------------------------------- XI I I
圖目錄----------------------------------------------------------------- XV I I
壹、緒言--------------------------------------------------------------- 1
貳、文獻回顧---------------------------------------------------------- 4
一、食品保存簡介---------------------------------------------- 4
二、食品保存劑之介紹------------------------------------- 6
(一)食品防腐劑之特性------------------------------------ 7
(二)食品防腐劑的來源------------------------------------ 8
(三)食品防腐劑之作用機制------------------------------ 35
三、常見之食品中毒污染菌------------------------------------ 37
(一)枯草(芽胞)桿菌 (Bacillus subtilis ) ---------------- 37
(二)大腸桿菌 (Escherichia coli)------------------ ------ 37
(三)黃桿菌屬(Flavobacterium) -------------------------- 38
(四)李斯特單胞菌(Listeria monocytogenes) ---------- 38
(五)綠膿桿菌(Pseudomonas aeruginosa) ---------- 38
(六)鼠傷寒桿菌( Salmonella typhimurium) -------- 39
(七)金黃色葡萄球菌(Staphylococcus aureus) --------- 39
(八)腸炎弧菌(Vibrio parahaemolyticus) --------------- 39
四、竹筍簡介---------------------------------------------------- 40
(一)台灣食用竹筍------------------------------------------ 40
1.食用竹筍的種類與分佈--------------------------- 40
2.竹筍之加工------------------------------------------ 41
參、材料與方法--------------------------------------------------------- 46
一、試驗材料----------------------------------------------------- 46
1.竹筍材料--------------------------------------------------- 46
2.試驗菌株---------------------------------- ---------------- 46
3.培養基及試藥--------------------------------------------- 46
二、實驗方法------------------------------------------------------ 48
(一)不同品種竹筍之篩選--------------------------------- 48
1.樣品之製備-------------------------------------------- 48
2.利用單一溶劑萃取與製備-------------------------- 48
(二)篩選最佳萃取條件------------------------------------ 51
1.以不同有機溶劑萃取抑菌性之物質-------------- 51
2.以水為介質萃取竹筍之抑菌物質----------------- 51
(三)抗菌性物質的製備------------------------------------ 51
(四)菌種之保存與更新------------------------------------------ 52
(五)種菌之製備--------------------------------------------- 52
(六)抑菌試驗及抑菌特性之探討------------------------ 52
1.抑菌活性之測定------------------------------------ 52
2.溫度對竹筍水萃取液之抑菌活性之影響------ 52
3. pH值對竹筍水萃取液抑菌活性之影響------- 53
4.金屬離子對竹筍水萃取液抑菌活性之影響--- 53
5.不同稀釋濃度之竹筍水萃取液對其抑菌活性之影響------------------------------------------------ 53
6.食品添加劑對竹筍水萃取液之抑菌活性影響 53
(1)化學試劑-------------------------------------- 53
(2)安定劑----------------------------------------- 53
(3)抗氧化劑-------------------------------------- 53
7. 添加蔗糖脂肪酸酯(sugar-ester)對竹筍水萃取液抑菌活性之影響---------------------------- 55
8. 添加幾丁聚醣乳酸鹽類(chitosan-lcatate)對竹筍水萃取液之抑菌性影響------------------- 55
9. 貯存條件對竹筍水萃取液之抑菌作用影響- 55
10. 抑菌率測定--------------------------------------- 55
11.不同濃度之竹筍水萃取液及己二烯酸鉀(potassium sorbate) 對Staphylococcus aureus 生長之影響-------------------------------------------- 56
12.最小抑菌濃度 (minimun inhibitory concerntration,MIC)之測定--------------------- 56
13.與日本商品化食品保存劑抑菌性之比較---- 57
14.竹筍抑菌劑、sodium benzoate 及potassium sorbate對試驗菌株抑菌活性之比較---------- 57
15.統計分析(statistical analysis system,SAS) 57
(七)竹筍水萃取液抑菌模式之探討--------------------- 58
1.靜菌或殺菌作用之測定--------------------------- 58
2. 掃描式電子顯微鏡(Scanning electron microscopy, SEM)進行萃取液抑菌模式之初步探討---------------------------------------------- 58
(八)食品模式------------------------------------------------ 58
1.竹筍抑菌劑對市售奶茶之保存性之探討------ 58
2.竹筍抑菌劑對市售珍珠粉圓之保存性之探討 59
(九)竹筍水萃取物抑菌成分分離純化------------------ 59
1.膠體過濾色層分析(gel filtration chromatography)進行萃取液之初步分離純化 59
2.以Sephadex G-25過濾法測定竹筍中抑菌物質之分子量------------------------------------------ 60
3.Amberlite XAD-7管柱層析---------------------- 60
4. Sephadex G-25過濾層析------------------------- 61
5. Sephadex G-15過濾層析------------------------- 61
6.高效能液相層析(HPLC) ------------------------- 61
(十)竹筍抑菌物質之初步判定--------------------------- 62
1. 紅外光光譜分析---------------------------------- 62
2.氣相層析質譜儀(GC-MS) ------------------------ 62
肆、結果與討論-------------------------------------------------------- 63
一、不同品種竹筍萃取液對試驗菌株抑菌活性之探討-- 63
二、不同溶劑萃取液對試驗菌株抑菌活性之影響--------- 63
(一)不同溶劑萃取液對試驗菌株抑菌活性之影響--- 63
(二)以水為介質經不同萃取條件所得的萃取物對試驗菌株抑菌活性之影響---------------------------- 65
三、萃取液抗菌特性之探討------------------------------------ 69
(一)不同溫度處理對竹筍水萃取液抑菌活性之影響 69
(二)竹筍水萃取液經不同pH值處理對其抑菌活性之影響-------------------------------------------------- 77
(三)金屬離子對竹筍水萃取液抑菌活性之影響------ 81
(四)不同稀釋度之竹筍水萃取液對其抑菌活性之影響-------------------------------------------------------- 82
(五)食品添加劑對竹筍水萃取液抑菌活性之影響--- 87
1.化學試劑對竹筍水萃取液抑菌活性之影響--- 87
2.安定劑對竹筍水萃取液抑菌活性之影響------ 96
3.抗氧化劑對竹筍水萃取液抑菌活性之影響--- 97
(六)添加蔗醣脂肪酸酯 (sugar ester)對竹筍水萃取液抑菌活性影響-------------------------------------- 107
(七)添加幾丁聚醣乳酸鹽(chitosan lactate)對竹筍水萃取液之抑菌活性之影響-------------------------- 108
(八)貯存條件對竹筍水萃取液抑菌活性之影響------ 113
(九)不同濃度的竹筍水萃取液對試驗菌株之抑菌率 118
(十)不同濃度之竹筍水萃取液及己二烯酸鉀對S. aureus生長之影響------------------------------------ 128
(十一)不同竹筍水萃取物之最小抑制濃度(MIC) --- 129
(十二)不同品種竹筍水萃取液與日售商品化保存劑抑菌性之比較--------------------------------------- 136
(十三)自製竹筍抑菌劑、安息香酸鈉及己二烯酸鉀之抑菌比較------------------------------------------ 138
四、竹筍水萃取液抑菌模式之探討--------------------------- 143
(一)不同竹筍水萃取液對試驗菌株之靜菌或殺菌作用-------------------------------------------------------- 143
(二)半胱胺酸對竹筍水萃取液抑菌活性之影響------ 145
(三)利用描掃式電子顯微鏡 (Scanning Electron Microscopy, SEM)進行竹筍水萃取液之抑菌模式探討---------------------------------------------------- 145
五、食品模式------------------------------------------------------ 150
(一)自製竹筍抑菌劑與日售商品抑菌劑(3P-SA),對貯存於4℃之奶茶中總生菌數之影響-------------- 150
(二)自製竹筍抑菌劑與日售商品抑菌劑 (3P-SA)對常溫貯存珍珠粉圓之抑菌效果---------------------- 155
六、竹筍萃取液抑菌成分純化分離之探討------------------ 155
(一)膠體過濾色層分析(gel filtration chromatography)進行萃取液之初步分離純化 155
1.烏腳綠竹筍水萃取液之Sephadex G-25層析分離及其抑菌活性情形--------------------------- 158
2.麻竹筍水萃取液之Sephadex G-25層析分離及其抑菌活性情形--------------------------------- 158
3.綠竹筍水萃取液之Sephadex G-25層析分離及其抑菌活性情形--------------------------------- 160
(二)以Sephadex G-25層析法測定竹筍抑菌物質之分子量-------------------------------------------------- 160
(三)Amberlite XAD-7管柱層析-------------------------- 165
(四)Sephadex G-25過濾層析----------------------------- 166
(五)Sephadex G-15過濾層析----------------------------- 167
(六) A-Ⅲ-3之HPLC分析-------------------------------- 173
(七) A-Ⅲ-3抑菌物質之初步探討----------------------- 174
1.紅外光光譜分析(IR) ------------------------------ 174
2.氣相層析質譜分析(GC-MS) --------------------- 174
伍、結論------------------------------------------------------------------ 177
陸、參考文獻------------------------------------------------------------ 181
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