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研究生:謝碧娟
論文名稱:以微生物熱死滅動力學試驗平台探討Neosartorya fischeri之耐熱特性
論文名稱(外文):Using microorganisms thermal death dynamics test platform to explore the heat resistance of Neosartorya fischeri
指導教授:翁義銘翁義銘引用關係
學位類別:碩士
校院名稱:國立嘉義大學
系所名稱:食品科學系研究所
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
畢業學年度:102
語文別:中文
中文關鍵詞:Neosartorya fischeri耐熱性子囊孢子D值z值果汁
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Neosartorya fischeri為主要造成加工產品腐敗之耐熱性黴菌之一,其子囊孢子較常存在於土壤、腐敗有機物質、腐敗水果、蔬菜、其他植物產品及加工設備等。N. fischeri在某些菌株生長過程中會產生如fumitremorgins A與C以及verruculogen之黴菌毒素。
本研究利用熱死滅/衰變動力學裝置,來建立指標微生物N. fischeri熱死滅動力學參數(D值與z值)。以N. fischeri BCRC 32429作為殺菌之指標菌,其菌株之培養溫度與時間為30℃、30天,子囊孢子熱活化處理為80℃、10min,利用TDT管探討N. fischeri子囊孢子於不同加熱基質中,於83℃、87℃、91℃進行加熱,即可得到子囊孢子之熱死滅速率曲線圖,以計算各溫度下之菌株耐熱D值,並求出其z值。由實驗結果顯示,當加熱基質為100%果汁,其耐熱性為D83℃=25.22~30.91 min、D87℃=4.14~5.52 min、D91℃=1.06~1.33 min、z=5.8~5.9℃;20%果汁其耐熱性為D83℃=52.13~55.19 min、D87℃=10.83~11.53 min、D91℃=1.40~1.55 min、z=5.1~5.2℃;若子囊孢子於Mcilvaine’s 磷酸緩衝溶液pH 3.5中,其耐熱性為D83℃=32.79 min、D87℃=4.80 min、D91℃=0.50 min、z=4.5℃;在Mcilvaine’s 磷酸緩衝溶液pH 7.0其耐熱性為D83℃=53.93 min、D87℃=6.25 min、D91℃=0.91 min、z=4.5℃;當基質為去離子水其耐熱性為D83℃=50.71 min、D87℃=7.29 min、D91℃=0.80 min、z=4.4℃。
另外,也更進一步探討於果汁中添加不同有機酸,其對N. fischeri子囊孢子耐熱性之影響;其試驗條件溫度為85℃,且在不同時間間隔下進行加熱,以測定其耐熱特性,即可得到子囊孢子之熱死滅速率曲線圖,以計算出該溫度下之熱致死速率值(D-values)。實驗結果顯示,將N. fischeri子囊孢子接種於100%果汁中,若酸化劑為檸檬酸、乳酸與酒石酸,其耐熱性分別為D85℃=12.55~14.28 min、D85℃=11.13~13.76 min、D85℃=15.76~18.21 min;若加熱基質為20%果汁,以檸檬酸、乳酸與酒石酸作為酸化劑,其子囊孢子耐熱性分別為D85℃=22.07~25.21 min、D85℃=21.38~24.43 min、D85℃=32.09~35.24 min。
由上述實驗結果發現,N. fischeri會受到加熱基質組成分與特性不同,其對於子囊孢子之耐熱程度也有所差異。此外,N. fischeri之熱致死動力學則遵循一級反應動力學模式(first-order reaction)。
目錄………………………………………………………………..…..…I
圖目錄………………………………………………………………..…Ⅴ
表目錄………………………………………………………...........…...Ⅷ
中文摘要…………………………………………………………..........Ⅸ
英文摘要……………………………………………………............…..XI
壹、前言………………………………………………………………….1
貳、文獻整理…………………………………………………….…….....4
一、黴菌……………………………………………………………...…4
(一)黴菌之構造及分類………………………………..…………….4
(二)黴菌繁殖方法………………………………………..………….5
二、常見之食物媒介性黴菌…………………………………....……...8
(一)Neosartorya分類…………………………………………....…10
(二)Neosartorya形態構造特徵………………………………....…10
(三)Neosartorya存在與分布、其對人體之影響……………….…13
三、耐熱性黴菌所造成酸性產品腐敗問題…………..……………...14
(一)耐性黴菌之來源…………...………………………………..…14
(二)耐性黴菌引起之相關問題……………………………….....…16
(三)影響耐熱性黴菌生長及孢子萌發之因子…………………….19
(四)耐熱性黴菌之防治………………………………………….…19
四、微生物之耐熱性……………………………………………..…...23
(一)影響微生物之耐熱性因子………………………………….....23
(二)食品添加物對微生物耐熱性之影響……………………….…25
(三)微生物耐熱性測定………………………………………..…...29
五、食品熱殺菌……………………………………….………….…...30
(一)產品殺菌原理…………………………………………..……...30
(二)殺菌方法………………………………………..…..……….....31
(三)殺菌曲線之繪製圖…………………………..………………...31
(四)產品殺菌條件之訂定及產品殺菌值之選擇………………….32
(五)熱處理之生物效應……………………………………….……36
(六)加熱對食品成分之影響………………………….……………37
六、微生物之耐熱效應……………………………………………….39
(一)D值(D-value)………………………….……………………….39
(二)z值(z-value)…………………………………………….…….. 40
(三)F值(F-value) ……………………………………...……….…..41
七、微生物熱死滅動力學…………………………………………….43
(一)微生物之熱死規律-對數殘留定律……………………..…...43
(二)溫度對殺菌速率常數k之影響…………………………..……46
八、研究目的………………………………………………………….48
參、實驗架構………………………………………...………………….49
肆、材料與方法…………………………………………………………53
一、實驗材料………………………………………………………….53
(一)樣品…………………………………………..……………….. 53
(二)菌株……………………………………………..……………...53
(三)藥品………………………………..…………………………...53
(四)儀器………………………………………………………..…...54
二、藥品配置與樣品調配…………………………………………….56
(一)藥品配置……………………………………………..……….. 56
(二)果汁調配……………………………………………..……...... 56
三、成分分析檢測…………………………………………………….59
(一)pH值測定………………………….…………………………..59
(二)糖度測定………………………………………..……………...59
(三)食品酸度(acidity)之測定………………………..………….....59
四、Neosartorya fischeri試驗株之活化……………………………...61
五、Neosartorya fischeri子囊孢子懸浮液之製備…………………...61
六、Neosartorya fischeri子囊孢子性狀觀察及濃度計數…………...62
(一)菌體生長型態觀察……………..……………………………...62
(二)子囊孢子濃度之計數……………………………..…………...62
七、微生物熱死滅動力學試驗平台…………………………………64
八、Neosartorya fischeri子囊孢子於不同加熱基質之耐熱性測定67
九、於果汁中添加不同有機酸對Neosartorya fischeri子囊孢子之耐熱性試驗…………………………………………....…...……....68
十、Neosartorya fischeri子囊孢子數之檢測方法…………………...69
十一、微生物耐熱特性參數之計算………………………………….70
(一)D值(D-value)……………….……..…………………………...70
(二)z值(z-value)……………………………………………….…...72
伍、結果與討論…………………………………………………………74
一、熱活化處理之選擇……………………………………………….74
二、Neosartorya fischeri菌體生長與子囊孢子型態之觀察………...74
三、果汁之pH值、糖度(°Brix)與酸度(%)測定……………………75
四、Neosartorya fischeri子囊孢子加熱基質中,於不同溫度下之升溫曲線…………………………………………………………...75
五、不同加熱基質對Neosartorya fischeri子囊孢子耐熱性之影響76
六、於果汁中添加不同有機酸對Neosartorya fischeri子囊孢子耐熱性之影響………………………………………………………...82
七、以Neosartorya fischeri子囊孢子為指標菌生產酸性產品之熱殺菌條件評估……………………………………………………...89
陸、結論…………………………………………………………………91
柒、參考文獻…………………………………………………………..119
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