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研究生:梁正融
研究生(外文):Liang, Zheng-Rong
論文名稱:乳酸鏈球菌素及亞硫酸鹽對白蝦冷藏期間之品質影響
論文名稱(外文):The Effect of Nisin and Sulfite on the Quality of White Shrimp during Refrigerated Storage
指導教授:蕭心怡蕭心怡引用關係
指導教授(外文):Hsiao, Hsin-I
口試委員:張月櫻林仲聖潘崇良宋文杰
口試委員(外文):chungsl@mail.ypu.edu.tw
口試日期:2016-06-29
學位類別:碩士
校院名稱:國立臺灣海洋大學
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:2016
畢業學年度:104
語文別:中文
論文頁數:73
中文關鍵詞:白蝦碎冰亞硫酸鹽乳酸鏈球菌素加乘效應
外文關鍵詞:White shrimpFlake iceSulfiteNisinSynergistic effect
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蝦子在一般保存條件下,因多酚氧化酶酵素性黑變 (melanosis)、微生物孳生與脂質氧化,導致貯存期限十分短暫,以往添加亞硫酸鹽防止蝦頭黑變以及抑制微生物生長,但是業者經常濫用超出限量標準,再加上民眾逐漸開始重視化學添加物對人體帶來的影響,目前趨勢為開發天然物質及物理方法取代亞硫酸鹽。乳酸鏈球菌素 (nisin) 為Lactococcus lactis subsp. lactis 所產生之細菌素,可抑制細菌之生長,美國食品藥物管理局1988年正式將其列入 GRAS級之防腐劑,目前已普遍應用於食品之保存。因此本研究將冷藏太平洋白蝦 (Litopenaeus vannamei) 單獨添加碎冰、nisin發酵液、偏亞硫酸氫鈉 (sodium metabisulfite,SMS) 以及nisin發酵液與偏亞硫酸氫鈉混合使用,於1±1oC下貯藏10天結果如下:一、Nisin標準品對Streptococcus thermophilus最小抑制生長濃度為50 ppm,發酵液中有效nisin濃度為47.22 ppm;二、Nisin與20 mM EDTA混合溶液對三株革蘭氏陰性菌之抗菌效果由高至低依序為V. parahaemolyticus、P. fluorescens及Sal. enterica;三、添加偏亞硫酸氫鈉可減緩白蝦之pH值上升,且冷藏白蝦於1±1oC下最大貯存期限為8天;四、純碎冰以及碎冰添加偏亞硫酸氫鈉能延緩白蝦之黑變,且效果較添加nisin發酵液佳;五、偏亞硫酸氫鈉搭配nisin發酵液能有效抑制微生物生長,具有加乘效應 (synergistic effect)。綜合以上,未來可以將nisin用於取代部分亞硫酸鹽之添加,減少化學添加物用於白蝦之使用量。
Due to polyphenol oxidase enzyme melanosis, microorganism and lipid oxidation, shelf life of shrimp under normal storage conditions is very short. In general, sulfite was used in shrimp to prevent from enzymatic browning and inhibiting microbial growth. However, over used by food processors is the major concern. In addition to this, consumers now concern the impact of chemical additives on the human body. There is a growing trend using natural substances or physical methods to replace sulfite in food. Nisin is one kind of bacteriocins, which secreted from Lactococcus lactis subsp. Nisin can inhibit the growth of bacteria. The US Food and Drug Administration in 1988 officially had included it as GRAS level of preservatives. So far it has been widely used in food preservation. To achieve our goals, this study divided Pacific white shrimp (Litopenaeus vannamei) intro following groups: control group (Control), ice group (ICE), 12.5 g / L sodium metabisulfite (SMS) with flake ice group (ICE_S), nisin broth with flake ice group (ICE_N) and 6.25 g / L SMS mixed nisin broth with flake ice group (ICE_S_N) and stored in 10-day. Following results were obtained: First, the minimum inhibitory concentration of nisin broth standard to Streptococcus thermophilus is 50 ppm. seanwhile the the minimum inhibitory concentration of nisin in the nisin fermented broth was found to be 47.22 ppm. Second, the antibacterial effect of three Gram-negative bacteria of nisin mixed 20 mM EDTA solution from high to low was V. parahaemolyticus, P. fluorescens and Sal. enterica. Third, shrimp with flake ice added sodium metabisulfite can reduce the pH rise, and the maximum of storage was 8-day. Fourth, the flake ice with sodium metabisulfite group can retard the melanosis. Fifth, sodium metabisulfite mixed with nisin broth can inhibit the growth of microorganisms, in which synergistic effect was observed. Based on our findings, nisin could be used to replace part or all of sulfite in the future.
壹、 前言 7
1.1 研究動機 7
1.2 研究目的 8
貳、文獻整理 9
2.1水產品保鮮 9
2.1.1水產品保鮮物理性方法 9
2.1.2水產品保鮮化學性方法 10
2.1.3欄柵效應、加乘效應 10
2.2冷藏蝦之品質變化 10
2.2.1冷藏蝦之黑變 11
2.3水產品之鮮度指標 11
2.3.1 pH 值 11
2.3.2 ATP 相關化合物與K值 12
2.3.3總揮發性鹽基態氮 (Total volatile basic nitrogen, TVB-N) 12
2.3.4 三甲胺(Trimethylamine, TMA) 12
2.3.5總生菌數 (The aerobic plate count, APC) 13
2.3.6水產品常見之腐敗菌、病原菌 13
2.4 亞硫酸鹽的特性與用途 13
2.4.1亞硫酸鹽之毒性 14
2.4.2二氧化硫之毒性 15
2.4.3食品中亞硫酸鹽使用之調查 15
2.4.4各國對亞硫酸鹽之管理與法規 15
2.5乳酸菌與乳酸鏈球菌素 16
2.5.1乳酸菌 16
2.5.2乳酸鏈球菌素 16
參、研究材料 18
3.1實驗樣品 18
3.2實驗菌株 18
3.3實驗藥品 18
3.4培養基 18
3.5儀器設備 19
3.6實驗架構 20
肆、研究方法 21
4.1菌株之保存、活化、及生長曲線 21
4.1.1菌株之保存 21
4.1.2菌株之活化 21
4.1.3菌株之生長曲線 21
4.2 Nisin標準品、發酵液及EDTA溶液之配製 21
4.2.1乳酸鏈球菌素 (nisin) 標準品溶液之製備 22
4.2.2乳酸鏈球菌素 (nisin) 發酵液之製備 22
4.2.3 EDTA溶液之製備 22
4.3 Nisin之抗菌試驗 22
4.3.1 Nisin標準品及發酵液對指標菌洋菜孔洞擴散法抗菌活性之測試 22
4.3.2 Nisin標準品對三株革蘭氏陰性菌之抗菌測試 23
4.4白蝦品質變化之測定 23
4.4.1 TVB-N 23
4.4.2 pH值 24
4.4.3顏色變化 (CIE L*a*b*) 24
4.4.4 微生物檢測 24
伍、結果與討論 25
5.1乳酸鏈球菌素 (nisin) 標準品之抗菌活性 25
5.1.1三株革蘭氏陰性菌生長曲線之測定 25
5.1.2 Nisin對指標菌之抗菌效果及發酵液標定結果 25
5.1.3 Nisin與EDTA對革蘭氏陰性菌之抗菌效果 25
5.2以不同添加物處理之冷藏白蝦貯存期間之變化 26
5.2.1 TVB-N 26
5.2.2 pH值 27
5.2.3顏色變化 27
5.2.4微生物檢測. 28
陸、綜合討論 30
柒、結論 31
捌、參考文獻 32
玖、圖表 40
壹拾、附錄 64
A1食品添加物使用範圍及限量-第一類 防腐劑 64
A2食品添加物使用範圍及限量-第四類 漂白劑 (以亞硫酸鹽類為例) 69


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