臺灣博碩士論文加值系統

食品發酵為最古老的食品加工方法之一，發酵肉製品製作過程中，除了添加乳酸菌酛外，亦可能添加可產酸之物質，降低pH值。葡萄糖酸內酯 (glucono-δ-lacton, GDL) 與檸檬酸 (citric acid, CA) 常添加於發酵產品中，不僅可以增加食物風味及延長食品保存，GDL亦具有抑制病原菌微生物之效果，然而若添加過量時會造成產品質地崩解品質下降，因此，本試驗旨在探討添加不同比率之GDL與檸檬酸對泰式發酵香腸Nham之理化、微生物、質地、感官品評及及電泳結果之影響。結果顯示：第4天發酵完成後，各組pH值皆顯著下降 (P < 0.05)，貯存期間各組之pH值及水活性皆無顯著變化 (P > 0.05)。發酵完成後各組滴定酸度皆顯著上升 (P < 0.05)，貯存期間各組皆有持續緩慢上升之趨勢，對照組測值最低；添加GDL與檸檬酸之組別隨著添加量的提升，滴定酸度有上升之趨勢，但各組之間無顯著差異 (P > 0.05)。製程第0天時，各組之L*值與a*值皆無顯著差異，發酵完成後各組皆有上升之趨勢；發酵後以及貯存期間b*值無顯變化，且各組間無顯著差異 (P > 0.05)，貯藏期結束後各組皆有下降之趨勢。物性分析結果顯示，發酵完成後樣品之硬度、彈性、脆度及咀嚼性皆顯著上升 (P < 0.05)，黏合力第0及第4天間樣品之無顯著差異，貯存期間各組硬度皆無顯著變化，對照組之測值皆為最高，添加GDL與檸檬酸之組別測值較低。產品經發酵完成後，VBN值有顯著上升 (P < 0.05)，其中又以對照組測值最高，檸檬酸處理組次之，添加GDL比率越高者其VBN測值越低。非蛋白態氮方面，發酵完成後各處理組之非蛋白態氮皆顯著上升 (P < 0.05)，儲存期間CA01、GDL04與GDL08皆有持續上升之趨勢，其中又以CA01組上升最多；貯藏期間，對照組、GDL04+CA01與GDL08+CA01則有緩慢下降之趨勢。SDS-PAGE結果顯示，無論是否添加GDL及檸檬酸皆不影響發酵過程中樣品蛋白質之分解，發酵期間 (第2天) 各處理組之間肌凝蛋白重鏈有逐漸降解之趨勢，經發酵後各處理組45 kDa分子片段亦有逐漸降解之趨勢發酵完成總生菌數與乳酸菌數皆顯著上升 (P < 0.05)，但各組間無顯著差異 (P > 0.05)。品評結果部分，隨著貯存時間延長，各組之顏色、氣味、質地、風味、酸度、鹽度及總接受度評分皆無顯著差異 (P > 0.05)。綜觀上述，添加GDL與檸檬酸於Nham中對理化、微生物、物性、感官品評及電泳結果之表現並無顯著差異，未來應考量添加GDL與檸檬酸對於病原性微生物之作用，以生產安全衛生及品質優良之泰式發酵香腸。

Food fermentation is one of the oldest food processing and preservation methods. Glucono-δ-lacton (GDL) and citric acid have been applied in the meat industry to improve flavors and extend shelf life of fermented products. However, too excess addition of GDL might deteriorate the qualities of products. The objective of this study was to investigate the influence of adding different amounts of GDL and citric acid on the physicochemical, microbiological, textural, and sensory properties of Nham sausages which inoculated with Lactobacillus plantarum as a starter culture.
The results showed that the pH values decreased significantly (P < 0.05) after finishing of fermentation (day 4), whereas no significant difference in pH values and water activity between treatments was observed during storage. The acidity increased dramatically during fermentation and then increased steadily thereafter and the control had the lowest acidity. Addition higher levels of GDL and citric acid tended to have higher acidity but there was no significant difference between treatments (P > 0.05). The L* and a* values did not significantly differ at day 0 and tended to increase after finishing of fermentation. Less change was observed in b* value after fermentation and during storage. There was no significant difference on b* values between treatments (P > 0.05) and they tended to decrease after storage. For the texture profile, the hardness, springiness, fracturbility and chewiness of samples increased significantly after fermentation, yet the cohesiveness of samples did not significantly differ between days 0 and 4. The hardness of samples did not significantly differ during storage. The control had the highest hardness whereas the ones with addition of GDL and citric acid had lower hardness. The VBN values increases significantly during the fermentation (P < 0.05). The control had the highest VBN values, followed by the ones with addition of citric acid; the more with addition of GDL, the less of the VBN values. The counts of total viable bacteria and lactic acid bacteria counts increases rapidly during fermentation and there was no significant difference between treatments (P > 0.05). There was no significant difference on the sensory characteristics between treatments. The NPN values of all treatments increased rapidly during the fermentation, and the ones of GDL04+CA01 and GDL08+CA01 decreased during storage. For the SDS-PAGE, the intensity of the myosin heavy chain (MHC) band decreased thereafter. The actin band (around 45 kDa) also decreased in intensity after fermentation and storage. In conclusion, adding GDL and citric acid did not detriment the physicochemical, microbiological, and textural qualities of the Nham sausages inoculated with L. plantarum. Further studies should be addressed on the protectium against the pathogens base on the addition of GDL and citric acid in order to produce safe Nham sausage with better qualities.

誌謝. i
中文摘要 ii
Abstract iii
圖次. vii
表次. viii
附錄目次 ix
壹、 前言 1
貳、 文獻探討 2
一、發酵食品定義與起源 2
二、菌酛 (starter culture) 2
(一) 乳酸菌之定義與用途 2
(二) 添加菌酛目的 2
(三) 乳酸菌之發酵作用與食品中之運用 3
(四) L. plantarum之運用以及與生物胺之關係 3
三、發酵香腸 5
(一) 發酵香腸之加工特性 5
(二) 發酵香腸分類 5
(三) 菌酛在發酵香腸中之利用 5
四、泰式發酵香腸Nham 6
五、GDL之介紹 8
(一) GDL之結構與功能性 8
(二) GDL使用 8
六、檸檬酸之介紹 9
(一) 檸檬酸之結構與功能性 9
(二) 檸檬酸之使用 9
參、 材料與方法 11
一、原料肉 11
二、泰式發酵香腸Nham的配方與製備 11
三、乳酸菌酛之活化與製備 11
四、分析項目 13
(一) 產品失重率 (weight loss) 13
(二) 水分含量 (moisture content) 13
(三) 灰分 (ash) 13
(四) 粗脂肪 (crude fat) 13
(五) 粗蛋白 (crude protein) 13
(六) pH值 (pH value) 14
(七) 酸度 (titratable acidity) 14
(八) 色澤 (color) 14
(九) 水活性 (Water activity) 15
(十) 質地剖面分析 (Texture profile analysis, TPA) 15
(十一) 揮發性鹽基態氮 (Volatile basic nitrogen, VBN) 15
(十二) 總生菌數 (total plate count) 16
(十三) 乳酸菌數 (lactic acid bacteria counts, LAB) 16
(十四) 非蛋白態氮 (nonprotein nitrogen, NPN) 16
(十五) SDS-PAGE 17
(十六) 感官品評 (sensory tralnation) 18
(十七) 統計分析 18
肆、 結果與討論 19
一、失重率、一般成分、pH值及酸度 19
二、顏色 23
三、水活性 25
四、物性分析 25
五、揮發性鹽基態氮 29
六、總生菌數及乳酸菌數 31
七、非蛋白態氮 33
八、SDS-PAGE 35
十、感官品評 39
伍、結論 41
陸、參考文獻 42
柒、附錄 49

林書蓉。2008。不同蒸煮米或紅麴米之添加量對泰式發酵香腸於4±2 °C貯存期間品質之影響。國立中興大學動物科學系碩士論文。
林禮旺。2002。原料肉處理與熟成溫度對中式乾香腸品質之影響。國立屏東科技大學碩士論文。
涂榮珍。2002。陳年中式乾醃火腿表面黴菌菌元篩選及其於乾醃里脊肉塊之研究。國立台灣大學碩士論文。
陳明造，賴滋漢，許哲彰。2000。添加葡萄糖酸內酯與不同米飯類對肉品質特性和生物胺含量之影響。中國畜牧學會會誌。第29卷 第三期 255-264。
陳佩莉。2011。應用Lactobacillus plantarum及發酵米糠床製作中式半乾式香腸之品質特性。國立嘉義大學碩士論文。
陳慶源，黃崇真，邱雪惠，廖啟成。2007。乳酸菌之保健功效與產品開發。農業生技產業季刊。第11卷60-68。
郭俊欽。1989。發酵食品。徐氏基金會出版，台北市。
楊盛行。2007。基礎微生物學。第511、515頁。麥格羅希爾出版，藝軒總行銷，台北市。
衛生福利部食品藥物管理署。2013。食品添加物使用規範及限量暨規格標準。
劉英俊，汪金追，劉裕國。1996。最新微生物應用工業。第63頁。中央出版社，台北市。
蘇遠志。2001。應用微生物學。第13、431、432頁。華香園出版社。
Adams, M. R. and P. Marteau. (1995). On the safety of lactic acid bacteria from food. Internal Journal of Food Microbial, 27, 263-264.
AOAC 1990. Official Methods of Analysis. 15th ed. Association of official Analytical Chemistry, Virginia, U.S.A.
Astiasaran, I., R. Villanueva, and J. Bello. (1990). Analysis of proteolysis and protein insolubility during the manufacture of some varieties of dry sausage. Meat Science, 28, 111-117.
Bacus, J. N. and W. L. Brown. 1981. Use of microbial culture: meat products. Food Technology, 35(1), 74-98.
Barbut, S. (2006). Fermantation and chemical acidification of salami type products: Effect on yield, texture and microstructure. Journal of Muscle Food, 17,34-42.
Bartholomew, D. T. and T. N. Blumer. (1980). Effect of lactic acid bacteria on quality of country-style ham. Journal of Food Science, 45,426-429.
Blom, H., B. F. Hagen, B. O. Pedersen, A. L. Holck, and L. A. Axelsson. (1996). Accelerated production of dry fermented sausage. Meat Science, 43, 229-242.
Boulten, R. (1980). The realtionships between total acidity, titratable acidity and pH in wine. American journal of enology and viticulture, 31, 76-80.
Bruggen, K. A. (1994). Dry-sausages technology. I. Chicago: Meat Science School, Viskase Corporation.
Candogan, K., F. B. Wardlaw and J. C. Acton. (2009). Effect of starter culture on proteolytic changes during processing of fermented beef sausages. Food Chemistry, 116(3), 731-737.
Careri, C., A. Mangia, G. Barbieri, B. R. Virgili and G. Parolari. (1993). Sensory property relations to chemical data of Italian type dry-cured ham. Journal of Food Science, 58, 968-972.
Daly, C., M. La Chance, W. E. Sandine and P. R. Elliker. (1973). Control of staphylococcus aureus in sausage by starter cultures and chemical. Journal of Food Science, 38, 426-430.
DeMasi, T. W., F. B. Wardlaw, R. L. Dick and J. C. Acton. (1990). Non-protein nitrogen (NPN) and free amino acid contents of dry fermented and non-fermented sausages. Meat Science, 27, 1-12.
Demeyer, D., J. Hoozee and H. Mesdom. (1974). Specificity of lipolysis during dry sausage ripening. Journal of Food Science, 39, 293-296.
Eskeland, B. and J. Nordal. (1980). Nutritional evaluation of protein in dry sausage during the fermentation process with special emphasis on amino acid digestibility. Journal of Food Science, 45, 1153-1160.
FDA. (1976). Bacteriological analytical manual for foods. Food and Drug Administration Bureau of Food. U.S.A.
Feiner, G. (2006). Meat product handbook. Woodhead Publishing Limited, England.
Fernandez, M., J. A. Ordonez, J. M. Sruna, B. Herranz and L. Hoz. (2000). Accelerated riping of dry fermented sausage. Trends Food Science and Technology, 11, 201-209.
Fernández-López, J. (1998). Estudio del color por meÂtodos objetivos en sistemas modelo de pastas de embutidos crudo-curados. Ph.D. thesis, Universidad de Murcia. Spain.
Flores, J., J. R. Marcus, P. Nieto, J. L. Navarro and P. Lorenzo. (1997). Effect of processing conditions on proteolysis and taste of dry-cured sausages. Z Lebens Unters Forsch A, 204, 168-172.
Garcia de Fernando, G. D. and P. F. Fox. (1991). Study of the proteolysis during the processing of dry fermented pork sausage. Meat Science, 30, 367-383.
Georgantelis, D., I. Ambrosiadis, K. Panagiota, G. Blekas and S. A. Georgakis. (2007). Effect of rosemary extract, chitosan and α-tocopherol on microbiological parameters and lipid oxidation of fresh pork sausages stored at 4 C. Meat Science, 76, 172-181.
Gill, C. (1976). Substrate Limitation of Bacterial Growth at Meat Surfaces. Journal of Applied Bacteriology, 41, 401-410.
González-Fandos, E., B. Herrera and N. Maya. (2009). Efficacy of citric acid against Listeria monocytogenes attached to poultry skin during refrigerated storage. International Journal of Food Science and Technology, 44, 262-268.
Hagen, B. F., J. L. Berdague, L. Holck, H. Naes and H. Blom. (1996). Bacterial proteinase reduces maturation time of dry fermented sausages. Journal of Food Science, 61(5), 1024-1029.
Halloran, O. G. R., D. R. Troy, D. J. Buckley and W. J. Reville. (1997). The role of endogenous proteases in the tenderization of fast glycolysing muscle. Meat Science, 47, 187-200.
Hu, Y., W. Xia and X. Liu. (2007). Changes in biogenic amines in fermented silver crap sausage inoculated with mixed starter cultures. Food Chemistry, 104, 188-195.
Hughes, M. C., J. P. Kerry, E. K. Arendt, P. M. Kenneally, P. L. H. McSweeney and E. E. O’Neill. (2002). Characterization of proteolysis during of semi-dry fermented sausage. Meat Science, 62, 205-216.
Incze, K. (1998). Dry fermented sausage. Meat Science, 49(1), 169-177.
Jay, J. M. (1992). Modern food microbiology. New York: Chapman and Hall.
Johansson, G., J. L. Berdague, W. Larsson, N. Tran and E. Borch. (1994). Lipolysis, proteolysis and formation of volatile components during ripening of a fermented sausage with Pediococcus pentosaceus and Staphylococcus xylosus as starter cultures. Meat Science, 38, 203-218.
Juncher, D., C. S. Vestergaard, J. S. Jensen, C. J. Weber , G. Bertelsen and L. H. Skibsted. (2000). Effects of chemical hurdles on microbiological and oxidative stability of a cooked cured emulsion type meat product. Meat Science, 55, 483-491.
Kato, T. and T. Ohashi. (1987). Preparation of fermented sausage by lactic acid bacteria. Jap. J. Dairy. Food Science, 36(6), A343-A349.
Kato, T., K. Kanie, I. Shiga and Y. Sato. (1985). Preparation of Fermented sausage by lactic acid bacteria. Nippon Nogeikaku Kaishi, 59, 248-255.
Kato, T., T. Itaya, M. Nose, T. Tahara, M. Sugimoto and Y. Sato. (1990). Preparation of uncooked semi-fermented sausage by lactic acid bacteria. Nippon Shokuhin Kogyo Gakkaishi, 37(4), 248-255.
Klement, J. T., R. G. Cassens and O. R. Fenema. (1973). The association of protein solubility with physical properties in a fermented sausage. Journal of Food Science, 38, 1128-1131.
Klement, J. T., Cassens, R. G. and Fenema, O. R., (1974). The effect of bacterial fermentation on protein solubility in a sausage model system. Journal of Food Science, 39, 833-835.
Kongkiattikajorn, J. (2013). Potential of Fermented Sausage-Associated Lactic Acid Bacteria to Degrade Biogenic Amines During Storage.
Leistner, L. and L. Gorris. (1994) Food preservation by combined processes. European Commission-Directorate General XII, Science (p. 100), Research and Development, Final Report, FLAIR Concerted Action No. 7, Subgroup B.
Lücke, F. K. (2000). Utilization of microbes to process and preserve meat. Meat Science, 56, 105-115.
Lücke, F. K. (1998). Fermented Sausage. In (Wood, B. J. B., ed) Microbiology of Fermented Foods. 2nd ed vol. 2. P41-83. Blackie Academic and Professional, London.
Maijala, R. (1994). Histamine and tyramine production by a Lactobacillus strain subjected to external pH decrease. Journal of Food Protection, 57, 259-262.
Maijala, R. L., E. S. Herola, M. A. Aho and J. A. Hirn. (1993). The effect of GDL-induced pH decrease on the formation of biogenic amines in meat. Journal of Food Protection, 56, 125-129.
Mani-López, E. H. S. García and A. López-Malo. (2012). Organic acid as antimicrobials to control Salmonella in meat and poultry products. Food Research International, 45, 713-721.
Mateo, J., M. C. Maria, M. Dominguez and A. J. Maria. (1996). Taste compounds in Chorizo and their change during ripening. Meat Science, 44(4), 245-254.
Metaxopoulos, J., C. Genigeorgis, M. J. Fanelli, C. Franti and E. Cosma. (1981). Production of Italian dry salami: effect of starter culture and chemical acidulation on staphylococcal growth in salami under commercial manufacturing conditions. Applied and Environment Microbiology, 42(5), 863.
Miller, A. J., J. E. Call and R. C. Whiting. (1993). Comparison of organic acid salts for Clostridium botulinum Control in an Uncured Turkey Product. Journal of Food Protection, 56, 958-962.
Motarjemi, Y. (2002). Impact of small scale fermentation technology on food safety in developing countries. International Journal of Food Microbiology, 75, 213-229.
Paukatong, K. V. and S. Kunawasen. (2001). The Hazard Analysis and Critical Control Points (HACCP) generic model for the production of Thai pork sausage (Nham). Berliner Und Munchener Tierarztliche Wochenschrift, 114, 1-4.
Paul, R., S. Morgan and C. Hill. (2002). Preservation and fermentation: past, present and future. International Journal of Food Microbiology, 79, 3-16.
Pérez –Alvarez, J. A., E. S. Maria, F. Juana and A. Vicente. (1999). Physicochemical characteristics of Spanish-type dry-cured sausage. Food Research International, 32, 599-607.
Qvist, S., S. Katrine and Z. Peter. (1994). Growth suppression of Listeria monocytogenes in a meat product. Food Microbiology, 24, 283-293.
Riebroy, S., S. Benjakul, W. Visessanguan, U. Erikson and T. Rustad. (2009). Acid-induced gelation of natural actomyosin from Atlantic cod (Gadus morhua) and burbot (Lota lota). Food Hydrocolloids, 23(1), 26-39.
Roca, M. and K. Incze. (1990). Fermented sausage. Food Review International, 6(1), 91-118.
Scannell, A. G., P. M. Kenneally and E. K. Arendt. (2004). Contribution of starter cultures to the proteolytic process of a fermented non-dried whole muscle ham product. International Journal of food microbiology, 93(2), 219-230.
Shalaby, A. R. (1996). Significance of biogenic amines to food safety and human health. Food Research International, 29, 675-690.
Shchipunov, Y., S. Sarin, I. Kim and C. S. Ha. (2009). Hydrogels formed through regulated self-organization of gradually charging chitosan in solution of xanthan. Green Chemistry, 12, 1187-1195.
Sriphochanart, W. and W. Skolpap. (2010). Characterization of proteolytic effect of lactic acid Stahnk bacteria starter cultures on Thai fermented sausage. Food Biochnology, 24, 293-311.
Stahnke, L. H. (1995). Dried sausages fermented with Staphylococcus xylosus at different temperatures and with different ingredient levels - Part II. Volatile components. Meat Science, 2, 193-209.
Steinkruas, K. H. (1994). Nutritional significance of fermented foods. Food Research International, 27, 259-267.
Steinkruas, K. H. (1997). Classification of fermented foods: worldwide review of household fermented techniques. Food Control, 8, 311-317.
Spaziani, M., M. D. Torre and M. L. Stecchini. (2009). Changes of physicochemical, microbiological, and textural properties during ripening of Italian low-acid sausages. Proteolysis, sensory and volatile profiles. Meat Science, 81(1), 77-85.
Sun, W., C. Chun, Z. Mouming, Z. Qiangzhong and Y. Bao. (2011). Effect of composition and oxidation of protein on their solubility, aggregation and proteolytic susceptibility during processing of Cantonese sausage. Food Chemistry, 124, 336-341.
Talon, R., I. Lebert, A. Lebert, S. Leroy, M. Garriga, T. Aymerich. (2007). Traditional dry fermented sausages produced in small-scale processing units in Mediterranean countries and Slovakia. 1: Microbial ecosystems of processing environments. Meat Science, 77, 570-579.
Thai Industrial Standards Institute. (2003). Ministry of Industry. http://app.tisi.go.th/otop/pdf file/tcps146 46.pdf (In Thai).
Toldrá, F. (1998). Proteolysis and lipolysis in flavor development of dry-cured meat products. Meat Science, 49(1), 101-110.
Toldrá, F. and M. Flores. (1998). The role of muscle proteases and lipases in flavor development during the processing of dry-cured ham. Critical Reviews in Food Science and Nutrition, 38, 331-352.
Toldrá, F., M. C. Miralles and J. Flores. (1992). Protein extractability in dry-cured ham. Food Chemistry, 44, 391-394.
Toldrá, F, F., M. Cervero and C. Part. (1993). Porcine aminopeptidase activity as affected by curing agents. Journal of Food Science, 58(4), 724-726.
Varnam, A. H. and J. P. Sutherland. (1995). Fermented sausage. In Meat and Meat Product, p. 314-354. Chapman and Hall. London.
Verplaetse, A. (1994). Influence of raw meat properties and processing technology on aroma quality of raw fermented meat products. Proceedings of International Congress of Meat Science and Technology, 40, 45-65.
Verplaetse, A., M. De Bosschere and D. Demeyer. (1989). Proteolysis during dry sausage ripening. In Proceedings 35th international congress on meat science and technology, Copenhaegen, Denmark.
Visessanguan, W., S. Benjakul, A. Panya, C. Kittikun and A. Assavanig. (2005). Influence of minced pork and rind ratios on physico-chemical and sensory quality of Nham - a Thai fermented pork sausage. Meat Science, 69(2), 355-362.
Visessanguan, W., S. Benjakul, T. Smitinont, C. Kittikun, P. Thepkasikul and A. Panya. (2006). Changes in microbiological, biochemical and physico-chemical properties of Nham inoculated with different inoculum levels of Lactobacillus curvatus. LWT - Food Science and Technology, 39(7), 814-826.
Visessanguan, W., S. Benjakul, S. Riebroy and P. Thepkasikul. (2004). Changes in composition and functional properties of proteins and their contributions to Nham characteristics. Meat Science, 66, 579–588.
Wilson, D. J. (1982). Fermented and semi-dry sausage. American Meat Institute, 1-10.
Xiong, Y. L. (1994). Myofibrillar protein from different muscle fiber types: Implications of biochemical and functional properties in meat processing. Critical Reviews in Food Science and Nutrition, 34, 293-320.
Zapelena M. J., I. Astiasaran and J. Bello. (1999). Dry fermented sausage made with a protease from Aspergillus oryzae and/or a starter culture. Meat Science, 52, 403-409.