臺灣博碩士論文加值系統

Nham為常見泰國傳統發酵香腸之一，廣受泰國及周邊國家消費者喜愛。Nham之製作原料主要為豬肉、熟豬皮、熟米、大蒜、鹽及亞硝酸鹽或硝酸鹽等，製作時常會添加糖及辣椒以增添風味；原料混合後充填於塑膠腸衣或以香蕉葉包裹，於室溫發酵3-5天後食用，藉由乳酸菌作用使產品pH值下降。在台灣所販售之Nham多為小規模生產，原料及製作流程較無固定，且部分為生食方式食用，因此遭受食源性病原菌如Salmonella、Staphylococcus aureus及Listeria monocytogenes等之汙染風險極高，故本試驗擬自市售Nham發酵香腸中篩選乳酸菌，評估其抑菌能力，以做為未來製備發酵香腸菌元之可行性及評估其對產品特性之影響。
結果顯示，所篩選菌株於培養48小時後可降低pH值至5.0以下，但是對於Salmonella enterica及Escherichia coli O157:H7之抑菌能力表現不如預期。以肉汁作為培養基模擬發酵香腸之環境試驗結果顯示，其中C03菌株可於肉汁培養液中生長並有效地下降pH值，因此選擇此C03菌株進行基因序列鑑定及特性鑑定。經16S rDNA序列鑑定結果顯示，C03菌株為Fructobacillus durionis C03，API鑑定結果顯示，此菌株對於葡萄糖、果糖及蔗糖具有代謝能力，對乳糖則為微代謝能力。
接種商業發酵香腸常用菌元Lactobacillus plantarum ATCC 14971及Fb. durionis C03或混合兩者做為菌元以製作發酵香腸，探討此些菌株對於樣品產酸、色澤、物性以及感官品評之影響。結果顯示，發酵期間各組之pH值皆顯著下降 (P < 0.05)。發酵期間亮度有上升之趨勢，黃色度有下降趨勢而紅色度無顯著差異 (P > 0.05)，但各組間色度於發酵結束第4天皆無顯著差異 (P > 0.05)。總生菌數及乳酸菌數發酵期間皆顯著上升至約7.5 log CFU/g，且有相近之生長趨勢。物性分析結果顯示，樣品發酵期間之硬度及彈性顯著上升 (P < 0.05)，但咀嚼性及凝聚性則無顯著差異 (P > 0.05)。感官品評結果部分，各組間之顏色、氣味、風味、鹽度、硬度、酸度及總體接受度間皆無顯著差異 (P > 0.05)，其中以混合兩菌元之組別的色澤、氣味、風味及酸度有較高分數表現。綜觀上述，自市售泰式發酵香腸Nham篩選分離之乳酸菌儘管對於所選擇特定之病原菌抑制能力較有限，但做為菌元使用時所製備之發酵香腸產品理化特性及感官品質皆可被接受。

Nham is a traditional fermented meat product and is popular in Thailand and neighboring countrys. The main materials of Nham are ground pork, cooked shredded pork rind, garlic, cooked rice, salt and nitrite (or nitrate). Besides, chili or sugar is added in some receipt for enhancing flavor. The mixing materials are filled in plastic casing or banana leaves, and is further fermented in room temperature for 3-5 days where lactic acid bacteria (LAB) in sausage could decline pH of product during fermentation. In Taiwan, most Nham are produced on small scale without standard manufacturing process. Furthermore, Nham is mainly eaten in raw, which resulting in the risks of infection from food-borne pathogen, like Salmonella, Staphylococcus aureus and Listeria monocytogene are high. The objective of this study was to evaluate the potential of LAB isolated from Nham, a Thai fermented sausage, for application to starter culture of fermented sausage.
The results showed that the LAB isolated from commercial fermented sausage declined pH to 5.0 after inoculated 48 hours in regular medium. However, the results of inhibition test to common food-borne pathogen, including Escherichia coli O157:H7 ATCC35150 and Salmonella enterica ATCC 13076 were not well as we expected. The selected strains were cultured in meat broth medium for simulating environment in fermented sausage. Strain C03 could growth in meat broth and showed good acid producing ability than other strains. Thus, strain C03 was selected for the starter used in fermented sausage and identified as Fructobacillus durionis by using 16S rDNA sequencing. In the result of API test, Fructobacillus durionis C03 could metabolize glucose, fructose and sucrose.
For manufacturing fermented sausage, Lactobacillus plantarum ATCC 14971 and Fb. durionis C03 were inoculated and were evaluated their effects on acid, color, texture and sensory in fermented sausage. The results showed that pH values decreased significantly (P < 0.05) during fermentation. The L* value increased, b* value decreased during fermentation, but there was no significantly between treatments (P>0.05). The count of total viable bacteria and lactic acid bacteria counts increased to 7.5 log CFU/g, and showed same growth trend. The results of texture test showed that hardness and springiness increased significantly during fermentation (P < 0.05) but no significantly between treatments (P >0.05). There were no significantly difference in sensory (P > 0.05), but the treatment of mixed starter culture showed higher scores in color, flavor, taste and sourness. In conclusion, Fructobacillus durionis C03, which was isolated from commercial Thai fermented sausage, Nham, could be applied as starter culture for producing safety and stable fermented sausage.

致謝 i
中文摘要 ii
Abstract iii
目次 v
圖次 viii
表次 ix
附錄目次 x
壹、 前言 1
貳、 文獻探討 2
一、 乳酸菌特性 2
二、 應用於發酵之微生物分類 5
(一)、 乳酸菌類 5
(二)、 酵母菌及黴菌類 6
三、 乳酸菌代謝產物對於病原菌之抑菌作用 9
(一)、 有機酸類 9
(二)、 過氧化氫 (hydrogen peroxide, H2O2) 9
(三)、 細菌素 (bacteriocin) 10
四、 發酵香腸與乳酸菌 11
(一)、 發酵香腸製作 11
(二)、 發酵香腸之生物化學及微生物變化 14
(三)、 泰式發酵香腸 Nham 16
參、 材料與方法 17
一、 試驗架構 17
二、 市售發酵香腸菌株篩選 18
(一)、 市售發酵香腸來源 18
(二)、 篩選方法 18
三、 肉汁培養試驗 19
(一)、 原料肉 19
(二)、 肉汁製備 19
(三)、 菌株培養 19
(四)、 分析項目 19
四、 菌株鑑定及特性分析 21
(一)、 16S rDNA 序列鑑定 21
(二)、 API鑑定 22
(三)、 醣類代謝試驗 22
五、 發酵香腸試驗 23
(一)、 原料肉 23
(二)、 乳酸菌活化與接種 23
(三)、 泰式發酵香腸配方與製備 23
(四)、 分析項目 25
肆、 結果與討論 27
一、 自市售發酵香腸篩選乳酸菌特性之評估 27
二、 模擬發酵香腸試驗特性評估 27
三、 篩選菌株之鑑定及特性分析 31
(一)、 16S rDNA 序列鑑定 31
(二)、 API試驗 32
(三)、 醣類代謝試驗 35
四、 發酵香腸接種結果 38
(一)、 酸度及pH值 38
(二)、 顏色 40
(三)、 總生菌數及乳酸菌數分析 43
(四)、 物性分析-硬度、彈性、咀嚼性及凝聚性 45
(五)、 感官品評 48
伍、 結論 50
陸、 參考文獻 51
柒、 附錄 58

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