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研究生:邱如敏
研究生(外文):Ju-Min Chiu
論文名稱:麝香草酚與白藜蘆醇抑菌特性之探討
論文名稱(外文):Studies on the Antibacterial Activity of Thymol and Resveratrol
指導教授:邱義源邱義源引用關係翁義銘翁義銘引用關係
指導教授(外文):Robin Y.-Y. ChiouYih-Ming Weng
學位類別:碩士
校院名稱:國立嘉義大學
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:2004
畢業學年度:92
語文別:中文
論文頁數:69
中文關鍵詞:麝香草酚白藜蘆醇抑菌性
外文關鍵詞:ThymolResveratrolAntibacterial Activity
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麝香草酚 (thymol)為百里香精油之主成分,而百里香精油早被應用於防腐抗菌之用途;白藜蘆醇 (resveratrol, 3, 5 ,4’-trihydroxystilbene)係屬多酚類化合物,具有抗氧化、防癌及抑菌性。本研究針對麝香草酚與白藜蘆醇,經分別以不同濃度與大腸桿菌(Escherichia coli CCRC 10675 及E. coli O157:H7 CCRC 15374)以及金黃色葡萄球菌(Staphylococcus aureus CCRC 10780,CCRC 12655 及 CCRC 13962)混合1至40 min後,再利用稀釋法培養計數。結果顯示麝香草酚及白藜蘆醇對各測試菌株之抑制效果,皆隨處理濃度及時間的增加而明顯提高。單獨添加麝香草酚125 ppm或白藜蘆醇250 ppm時,處理至40 min時,對E. coli CCRC 10675及E. coli O157:H7 CCRC 15374兩者分別有0.16、0.78 log及0.26、0.18 log值之殺菌力,將兩者混合併用處理10至20 min時,則有1.02~3.31 log值之殺菌力,顯示麝香草酚與白藜蘆醇具有加成作用以增強其抑菌活性。對金黃色葡萄球菌之抑菌性而言,由S. aureus CCRC 10780、 CCRC 12655及CCRC 13962之抑菌結果顯示,單獨添加麝香草酚250 ppm處理5 min時約有2.88、0.1及0.04 log值之殺菌力,而單獨添加250 ppm之白藜蘆醇時,則無顯著殺菌力(0~0.5 log),然將兩者混合併用處理5 min時,即能完全抑制菌株之生長;以S. aureus CCRC 12655最為敏感,於混合併用處理1 min時,即完全被抑制其生長,可知混合併用組之抑菌效力優於單獨添加麝香草酚或白藜蘆醇者,顯示具有加成作用而增強其抑菌活性。應用於生鮮食品時,將苜宿芽與洋菇切片經麝香草酚或白藜蘆醇之乙醇溶液浸泡處理後,於4℃儲存期間測定總生菌數生長之變化。結果顯示,麝香草酚及白藜蘆醇對洋菇與苜宿芽之抑菌活性,皆隨處理濃度的增加而明顯提高,以500 ppm所處理者效果最好。針對苜宿芽而言,以麝香草酚500 ppm處理30 min並於4℃儲存至48 h時,有>1 log之抑菌效力,以相同濃度之白藜蘆醇處理苜宿芽約有0.33 log 之抑菌效力;對洋菇而言,當以麝香草酚或白藜蘆醇500 ppm處理並於4℃儲存48 h時,皆有>3 log之抑菌效力。就相同材料而言,麝香草酚之抑菌活性稍大於相同濃度之白藜蘆醇,但此二成分僅以125 ppm之濃度處理30 min,經4℃儲存48 h後,洋菇及苜宿芽之總生菌數皆低於或近於以去離子水所處理之控制組。顯示麝香草酚於較低之濃度250 ppm時,即能夠達到一定程度之抑菌活性,以及有效抑制4℃儲存洋菇及苜蓿芽之微生物存活,具有進一步發展應用之潛力。
Thymol, a major essential oil component of thyme and thyme oil, has long been used as an effective antimicrobial agent. Resveratrol (3,5,4’-trihydroxystilbene), one of the natural phenolic compounds, has been found as an antioxidative, anti-cancer and antimicrobial compound. Antibacterial activities of thymol and resveratrol against Escherichia coli CCRC 10675 and E. coli O157:H7 CCRC 15374, and Staphylococcus aureus CCRC 10780, CCRC 12655 and CCRC 13962, were tested in liquid medium. Surrvial of bacteria was monitered by broth dilution method and subsequent population enumeration. An effective retardation of the bacterial growth resulted from supplemention of thymol in the media at concerntrations of 125 ppm or resveratrol at 250 ppm for 40 min. The population decreases were 0.16 and 0.78 log for E. coli O157:H7 CCRC 15374 and 0.26 and 0.18 log for CCRC 10675, respectively. Incubated in medium with 125 ppm thymol plus 250 ppm of resvertrol for 10 and 20 min, the population of E. coli decreased by 1.02 to 3.31 log, indicating that a synergist effect was resulted from combination of thymol and resvertrol. When the S. aureus strains were subjected to treatment with 250 ppm thymol for 5 min, the population of CCRC 10780, CCRC 12655 and CCRC 13962 decreased by 2.88, 0.1 and 0.04 log, respectively. When the strains were treated with 250 ppm resveratrol for 1 and 5 min, the population decreased by only 0 to 0.51 log. When 250 ppm thymol and 250 ppm resveratrol were combined for 5 min of treatment, no growth was obserred. For CCRC 12655, S. aureus CCRC 12655 did not grow after 1 min of treatment and all test strains did not grow after 5 min of treatment. A synergist effect between thymol and resveratrol against S. aureus growth was demonstrated. When fresh mushroom slides and alfalfa sprouts were subjected to treatments with thymol or resveratrol at various concentrations for various times and followed by storage at 4 ℃ and microbial population enumeration, the total viable populations decreased with an increase of concentration or increase of treatment time. When alfalfa sprouts were treated with 500 ppm thymol or 500 ppm resveratrol for 30 min and stored at 4 ℃ for 48 h, the viable population decreases were larger than 1 log and 0.33 log, respectively. For the case of mushroom slides, after treatment with 500 ppm thymol or 500 ppm resveratrol for 30 min and stored at 4℃ for 48 h, the population decreases were larger than 3 log. In comparison at the same concentration level, the antibacterial effect of thymol was slightly more effective than was resveratrol. Application of thymol and resveratrol in practical uses are of perspective and potency.
目 錄
頁次
中文摘要………………………………………………………………..Ⅰ
英文摘要………………………………………………………………..Ⅲ
謝誌……………………………………………………………………..Ⅴ
表次……………………………………………………………………..Ⅵ
圖次……………………………………………………………………..Ⅷ
壹、 前言……………………………………………………………..1
貳、 文獻回顧………………………………………………………..4
一、 食品保存劑簡介………………………………………………..4
(一) 食品保存劑之特性……………………………………………..4
(二) 食品保存劑之抑菌作用機制…………………………………..4
(三) 食品保存劑對微生物生長之抑制形式………………………..6
二、 天然抗菌物質…………………………………………………..8
(一) 植物殺菌素………………………………………………….….8
(二) 有機酸類…………………………………………………….….8
(三) 植物精油………………………………………………………..8
(四) 酚類及其相關化合物…………………………………………..9
三、 抗細菌活性之試驗方法………………………………………12
(一) 瓊脂稀釋法………………………………………….…….12
(二) 液態培養基稀釋法………………………………………..13
(三) 濾紙片擴散法……………………………………….…….13
四、 抗菌成分之相關研究…………………………………………13
(一) Oleuropein…………………………………………………13
(二) Oleoresins…………………………………………………14
(三) Thymol及carvacrol……………………………………….14
(四) Bornel……………………………………………………...15
(五) Eugenol 及 cinnamic aldehyde…………………….…….16
(六) Linalool 及chavicol………………………………………17
(七) Vanillin…………………………………………………….17
(八) Terpenes…………………………………………………...18
五、 麝香草酚之研究………………………………………………20
(一) 簡介………………………………………………….…….20
(二) 抑菌性……………………………………………….…….20
(三) 抑菌機制…………………………………………….…….21
六、 白藜蘆醇之研究………………………………………………22
(一) 特性……………………………………………………….22
(二) 抗氧化性………………………………………………….22
(三) 抑菌性…………………………………………………….24
七、 食品污染菌簡介……………………………………………..25
(一) 金黃色葡萄球菌……………………………………….….25
1. 特性…………………………………………………….….25
2. 食品中毒……………………………………………….….25
(二) 大腸桿菌…………………………………………….....….27
1. 特性……………………………………………….……….27
2. 病原性大腸桿菌之分類…………………………..……....27
(1) 腸內病原性大腸桿菌……………………..……..27
(2) 腸管侵襲性大腸桿菌………………………..…..27
(3) 腸內產毒型大腸桿菌…………………………..28
(4) 腸內出血型大腸桿菌………………………..…..28
3. 食品中毒……………………………………...……..…….28
參、 麝香草酚與白藜蘆醇抑制金黃色葡萄球菌及大腸桿菌生長探討……………………………………………………………….30
一、 材料與方法……………………………………………………30
(一) 試驗材料…………………………………………………..30
1. 菌種來源………………………………………………......30
2. 培養基………………………………………………...….. 30
3. 化學試藥…………………………………………………..30
(二) 試驗方法…………………………………………………..32
1. 麝香草酚之抑菌活性……………………………………..33
2. 白藜蘆醇之抑菌活性…………………………………..…33
3. 白藜蘆醇與麝香草酚併用之抑菌效果之探討………......33
二、 結果與討論……………………………………………………35
(一) 不同濃度麝香草酚之抑菌活性……..……………………......34
(二) 不同濃度白藜蘆醇之抑菌活性………………………..……..37
(三) 白藜蘆醇與麝香草酚併用之抑菌活性……………………....39
肆、 麝香草酚與白藜蘆醇對新鮮洋菇與苜宿芽總生菌數生長之影 響…………………………………………………………………………….48
一、 材料與方法……………………………………………………48
(一) 試驗材料………………………………………………….…..48
1. 食品………………………………………………………..48
2. 培養基……………………………………………………..48
3. 化學試藥…………………………………………………..48
4. 麝香草酚及白藜蘆醇溶液之配製………………………..48
(二) 試驗方法………………………………………………….…..48
1. 對新鮮苜蓿芽總生菌數之測定…………………………..48
2. 對新鮮洋菇總生菌數之測定…………………………….49
二、 結果與討論……………………………………………………50
(一) 麝香草酚與白藜蘆醇對苜宿芽總生菌數生長之影響…..…..50
1. 麝香草酚對苜宿芽總生菌數生長之影響………………..50
2. 白藜蘆醇對苜宿芽總生菌數生長之影響………………50
(二) 麝香草酚與白黎蘆醇對新鮮洋菇切片總生菌數生長之影響52
1. 麝香草酚對新鮮洋菇切片中總生菌數生長之影響…….52
2. 白藜蘆醇對新鮮洋菇切片中總生菌數生長之影響…….52
3. 麝香草酚添加抗壞血酸,對新鮮洋菇色澤及總生菌數生長之影響…………………………………………………….56
伍、參考文獻…………………………………………………………...62
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