臺灣博碩士論文加值系統

本研究的目的旨在利用Flavourzyme酵素處理(FT)與Alcalase酵素處理(AT)豬後腿肉開發出具有特殊風味的調味物質。選擇 FT和AT的作用條件分別為pH 7.0、50 ºC和 pH 8.5、 60 ºC，酵素濃度則有1、2和5% 。結果顯示FT組和AT組水解8小時後，其胺基態氮量分別為168和98 mg%，而水解率則分別為49.3和28.45%。依上述之pH值與溫度條件對水解率和風味評估之影響，被認為是最適合的酵素水解條件。
蛋白質膠片電泳顯示，酵素具有良好的分解效果，蛋白質能被分解成較小的蛋白質片段和胜肽。FT組的總游離胺基酸較AT組高，且具有較高酸味和鮮味的胺基酸如天門冬胺酸和麩胺酸含量高，FT組分別為121.51和233.49 mg/100g，而AT組則為33.31和87.23 mg/100g。由於AT組的嚐味性較低，故先以Flavourzyme組作用完後再添加Alcalase進一步作用而形成另一個處理組(FAT)，供作另一種調味方式。將水解液進行噴霧與冷凍乾燥後，將以產品的水溶性和吸水性為指標作為將來乾燥方法之依據。在水溶性方面，冷凍乾燥的AT組最高，而冷凍乾燥的FAT組則是吸水性效果最高，兩者分別是92.4%和4.23。將FT組進一步調味，加入3%葡萄糖經過90ºC加熱90分鐘，再添加0.7%氯化鈉、0.5% IMP+ GMP和0.05%的麩胺酸鈉。最後進行品評試驗，結果發現，水解液經過調味後的品評分數與市售的商品並無顯著差異(P>0.05)。
綜上所述，本研究利用豬後腿肉水解液經過調味後，可製造出具有特殊風味物質，且接受度與市售商品無顯著差異。將此水解液當做基質可再進一步作成其他風味的調味料供添加於速食麵中。

The purpose of this study was focused on the flavor enhancing property of the hydrolysates and to develop a special flavor seasoning. There were two enzyme treatments, Flavourzyme-treatment (FT) and Alcalase-treatment (AT). The conditions for the hydrolysis of enzymes were pH 7.0, 50ºC for Flavourzyme and pH 8.5, 60ºC for Alcalase, and the enzyme concentrations were 1, 2, and 5% (w/w) respectively. The 8 hours FT and AT treated groups had 168 and 98 mg% of amino nitrogen respectively and the degrees of hydrolysis were 49.3 and 28.45% respectively. These two conditions were considered to be the optimal condition for hydrolysis.
The SDS-PAGE of the hydrolysates in FT and AT showed that proteins were peptides and other nitrogen-contained fragments, for which free amino acids were also analyzed. The amounts of total free amino acids were higher in FT. Aspartic acid and glutamic acid, which had sour and umami taste, were 121.51 and 233.49 mg/100g in FT and 33.31 and 87.23 mg/100g in AT, respectively. To improve the flavor, FT was treated with Alcalase to develop another treatment (FAT). Hydrolysates were spray-dried and freeze-dried in order to make commercial products, and their solubility index (WSI) and absorption index (WAI) were tested. AT showed the best WSI (92.4%) in freeze-dried group, and FAT had the highest WAI (4.23) in freeze-dried group. The Flovourzyme hydrolysate which were added 3% of glucose and heated for 90ºC, 90min, then with another addition of 0.7% of salt, 0.5% of IMP+GMP, organic acids, 0.05% monosodium glutamate. Sensory evaluation data showed that after modulation, the hydrolysates had scores close to the commercial product.
In conclusion, this study was conducted to make pork protein hydrolysates that can supply unique flavor in order to develop a seasoning with special taste. The FT hydrolysates after modulation had acceptable flavor. It could be further modified and used in instant food.

目錄
頁次
中文摘要............................................................................................................................i
英文摘要...........................................................................................................................ii
第一章 前言............................................................................................................................1
第二章 文獻回顧............................................................................................................................2
2.1 豬肉的消費現況和營養價值............................................................................................................................2
2.2 動物死後肌肉之化學變化............................................................................................................................5
2.3 豬肉的呈味成分...........................................................................................................................12
2.4 調味料的製造...........................................................................................................................17
2.5 風味添加物的種類和應用...........................................................................................................................21
第三章 材料與方法...........................................................................................................................25
3.1 實驗材料...........................................................................................................................25
3.2 水解液分析項目...........................................................................................................................27
3.3 電泳檢定法...........................................................................................................................31
3.4 乾燥方法與感官品評...........................................................................................................................34
3.5 統計分析...........................................................................................................................34
第四章 結果與討論...........................................................................................................................36
4.1 最佳水解條件之探討...........................................................................................................................36
4.2 水解液之理化性狀分析...........................................................................................................................44
4.3 水解液的調味與感官品評...........................................................................................................................54
第五章 結論...........................................................................................................................58
第六章 參考文獻...........................................................................................................................59
第七章 作者小傳...........................................................................................................................67

中國國家標準。1982。水分檢驗法CNS 2770-3。經濟部中央標準局。
農業統計。2006。行政院農業委員會。
台灣農業年報。2007。行政院農業委員會。
李秀、賴茲漢。1992。食品分析與檢驗，pp. 151-182。富林出版社，台北市。
卓雅怡。2006。鯖魚罐頭蒸煮液作為粉末調味料基質與魚醬油之探討。國立海洋
大學食品科學所碩士論文。基隆。
凃榮珍。2003。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。
國立臺灣大學畜產學研究所碩士論文。台北。
陳明造。2000。肉品加工理論與應用。藝軒圖書出版社，台北市。
黃加成。1993。應用乳酸菌於中式香腸之研究。國立臺灣大學畜產學研究所博士
論文。台北。
蘇和平。1982。自淘汰蛋雞水解物製造複合調味料基質之研究。國立臺灣大學畜
產學研究所碩士論文。台北。

Anderson, R. A., H. F. Conway, V. F. Pfeifer and E. L. Griffin. 1969. Gelatinization of
corn grits by roll and extrusion-cooking. Cereal Sci. Today. 14(1):11-12.

A.O.A.C. 1990. Official methods of analysis. 928.08 Nitrogen in meat：Kjeldahl
method. 15th ed. Association of Official Analytical Chemists, Washington, D. C.

Arai, S., M. Noguchi, S. Kurosawa, H. Kato and M. Fujimaki. 1970. Applying proteolytic enzymes on soybean .6. Deodorization effect of aspergillopeptidase-a and debittering effect of aspergillus acid carboxypeptidase. J. Food Sci. 35: 392-396.
Arihara, K. 2006. Strategies for designing novel functional meat products. Meat Sci. 74: 219-229.
Aristoy, M. C. and F. Told’ra. 1991. Deproteinization techniques for HPLC amino-acid-analysis in fresh pork muscle and dry-cured ham. J. Agr. Food Chem. 39: 1792-1795.
Ashie, I. N. A., T. L. Sorensen and P. M. Nielsen. 2002. Effects of papain and a microbial enzyme on meat proteins and beef tenderness. J. Food Sci. 67: 2138-2142.
Benito, M. J., J. J. C''ordoba, M. Alonso, M. A. Asensio and F. Nunez. 2003a. Hydrolytic activity of penicillium chrysogenum pg222 on pork myofibrillar proteins. Int. J. Food Microbiol. 89: 155-161.
Benito, M. J., M. Rodriguez, R. Acosta and J. J. C''ordoba. 2003b. Effect of the fungal extracellular protease epg222 on texture of whole pieces of pork loin. Meat Sci. 65: 877-884.
Bradford, M. M. 1976. Rapid and sensitive method for quantitation of microgram quantities of protein utilizing principle of protein-dye binding. Anal. Biochem. 72: 248-254.
Carnegie, P. R., M. G. Collins and M. Z. Ilic. 1984. Use of histidine dipeptides to estimate the proportion of pig meat in processed meats. Meat Sci. 10: 145-154.
Dear, N., K. Matena, M. Vingron and T. Boehm. 1997. A new subfamily of vertebrate calpains lacking a calmodulin-like domain: Implications for calpain regulation and evolution. Genomics 45: 175-184.
Djenane, D., L. Martinez, A. Sanchez-Escalante, J. A. Beltran and P. Roncales. 2004. Antioxidant effect of carnosine and carnitine in fresh beef steaks stored under modified atmosphere. Food Chem. 85: 453-459.
Fairweathertait, S. J. 1989. Iron in food and its availability. Acta. Paediatr Scand: 12-20.
Feiner, G. 2006. Meat products handbook Practical science and technology. Woodhead Publishing Limited.
FitzGerald, R. J. and G. O''Cuinn. 2006. Enzymatic debittering of food protein hydrolysates. Biotechnol. Adv. 24: 234-237.
Goll, D. E., Y. Otsuka, P. A. Nagainis, J. D. Shannon, S. K. Sathe and M. Muguruma. 1983. Role of muscle proteinases in maintenance of muscle integrity and mass. J. Food Biochem. 7: 137-177.
Goll, D. E., V. F. Thompson, H. Q. Li, W. Wei and J. Y. Cong. 2003. The calpain system. Physiol. Rev. 83: 731-801.
Hopkins, D. L. and J. M. Thompson. 2001a. Inhibition of protease activity 2. Degradation of myofibrillar proteins, myofibril examination and determination of free calcium levels. Meat Sci. 59: 199-209.
Hopkins, D. L. and J. M. Thompson. 2001b. Inhibition of protease activity. Part 1. The effect on tenderness and indicators of proteolysis in ovine muscle. Meat Sci. 59: 175-185.
Hughes, M. C., A. Healy, P. L. H. McSweeney and E. E. O''Neill. 2000. Proteolytic specificity of cathepsin d on bovine f-actin. Meat Sci. 56: 165-172.
Hughes, M. C., E. E. O''Neill, P. L. H. McSweeney and A. Healy. 1999. Proteolysis of bovine f-actin by cathepsin b. Food Chem. 64: 525-530.
Hunt, J. R., S. K. Gallagher, L. K. Johnson and G. I. Lykken. 1995. High-meat versus low-meat diets - effects on zinc-absorption, iron status, and calcium, copper, iron, magnesium, manganese, nitrogen, phosphorus, and zinc balance in postmenopausal women. Am J. Clin. Nutr. 62: 621-632.
Ishizaki, S., T. Tachihara, H. Tamura, T. Yanai and T. Kitahara. 2005. Evaluation of odour-active compounds in roasted shrimp (sergia lucens hansen) by aroma extract dilution analysis. Flavour Frag. J. 20: 562-566.
Kerry, J., J. Kerry, and a. D. Ledward. 2002. Meat processing Improving quality. CRC Press LLC.
Klompong, V., S. Benjakul, D. Kantachote and F. Shahidi. 2007. Antioxidative activity and functional properties of protein hydrolysate of yellow stripe trevally (selaroides leptolepis) as influenced by the degree of hydrolysis and enzyme type. Food Chem. 102: 1317-1327.
Koohmaraie, M., S. C. Seideman, J. E. Schollmeyer, T. R. Dutson and J. D. Crouse. 1987. Effect of postmortem storage on ca++-dependent proteases, their inhibitor and myofibril fragmentation. Meat Sci 19: 187-196.
Lahl, W. J. and S. D. Braun. 1994. Enzymatic production of protein hydrolysates for food use. Food Technol-Chicago 48: 68-71.
Lawrie, R. and D. Ledward. 2007. Lawrie''s meat science, seventh edition. Woodhead Publishing Limited and CRC Press LLC.
Lee, J. Y., H. D. Lee and C. H. Lee. 2001. Characterization of hydrolysates produced by mild-acid treatment and enzymatic hydrolysis of defatted soybean flour. Food Res. Int. 34: 217-222.
Luh, B. S. 1995. Industrial-production of soy-sauce. J. Ind. Microbiol. 14: 467-471.
Matsukura, U., A. Okitani, T. Nishimuro and H. Kato. 1981. Mode of degradation of myofibrillar proteins by an endogenous protease, cathepsin-l. Biochim. Biophys. Acta. 662: 41-47.
Melendo, J. A., J. A. Beltran, I. Jaime, R. Sancho and P. Roncales. 1996. Limited proteolysis of myofibrillar proteins by bromelain decreases toughness of coarse dry sausage. Food Chem.57: 429-433.
Mottram, D. S. 1998. Flavour formation in meat and meat products: A review. Food Chem. 62: 415-424.
Moya, V. J., M. Flores, M. C. Aristoy, and F. Told’ra. 2001. Pork meat quality affects peptide and amino acid profiles during the ageing process. Meat Sci. 58: 197-206.
Nakadai, T., S. Nasuno and N. Iguchi. 1973. Purification and properties of acid carboxypeptidase-iv from aspergillus-oryzae. Agr. Biol. Chem. Tokyo 37: 1237-1251.
Nishimura, T., A. Hattori and K. Takahashi. 1996. Relationship between degradation of proteoglycans and weakening of the intramuscular connective tissue during post-mortem ageing of beef. Meat Sci. 42: 251-260.
Nollet, L. M. L., and F. Told’ra. 2006. Advanced technologies for meat processing. CRC PressTaylor & Francis Group.

Okitani, A., T. Nishimura and H. Kato. 1981. Characterization of hydrolase-h, a new muscle protease possessing aminoendopeptidase activity. Eur. J. Biochem. 115: 269-274.
Ramezani, R., M. Aminlari and H. Fallahi. 2003. Effect of chemically modified soy proteins and ficin-tenderized meat on the quality attributes of sausage. J. Food Sci. 68: 85-88.
Rodrigueznunez, E., M. C. Aristoy and F. Told’ra. 1995. Peptide generation in the processing of dry-cured ham. Food Chem. 53: 187-190.
Saha, B. C. and K. Hayashi. 2001. Debittering of protein hydrolyzates. Biotechnol. Adv. 19: 355-370.
Schwartz, W. N. and J. W. C. Bird. 1977. Degradation of myofibrillar proteins by cathepsins b and d. Biochem. J. 167: 811-815.
Schweigert, B. S. and B. J. and Payne. 1956. A summary of the nutrient content of meat. American Meat Institute Foundation Bulletin 30.
Sharp, J. G. 1963. Aseptic autolysis in rabbit and bovine muscle during storage at 37 degrees. J. Sci. Food Agr. 14: 468-472.
Skov, A. R., S. Toubro, B. Ronn, L. Holm and A. Astrup. 1999. Randomized trial on protein vs carbohydrate in ad libitum fat reduced diet for the treatment of obesity. Int. J. Obesity 23: 528-536.
Soldo, T., I. Blank and T. Hofmann. 2003. (+)-(s)-alapyridaine - a general taste enhancer? Chem Senses 28: 371-379.
Solms, J. 1969. Taste of amino acids, peptides, and proteins. J Agr Food Chem 17: 686-690.

Suyama, M. and T. Shimizu. 1982. Buffering capacity and taste of carnosine and its methylated compounds. B. Jpn. Soc. Sci. Fish 48: 89-95.
Tikk, M., K. Tikk, M. A. Torngren and L. Meinert. 2006. Development of inosine monophosphate and its degradation products during aging of pork of different qualities in relation to basic taste and retronasal flavor perception of the meat. J. Agr. Food Chem. 54: 7769-7777.
Told’ra, F. 1998. Proteolysis and lipolysis in flavour development of dry-cured meat products. Meat Sci. 49: S101-S110.
Told’ra, F., M. C. Miralles and J. Flores. 1992. Protein extractability in dry-cured ham. Food Chem. 44: 391-394.
Uhlig, H. 1998. Industrial enzymes and their applications. Wiley-interscience publication, New York.
van Boekel, M. A. J. S. 2006. Formation of flavour compounds in the maillard reaction. Biotechnol. Adv. 24: 230-233.
Whitaker, R. J., A. G. J. Voragen and D. W. S. Wong. 2003. Handbook of food enzymology. Marcel Dekker,Inc. New York.
Wierbicki, E., L. E. Kunkle, V. R. Cahill and F. E. Deatherage. 1954. The relation of tenderness to protein alterations during post mortem aging. Food Technol-Chicago 8: 506-511.
Zheng, X. Q., L. T. Li, X. L. Liu, X. J. Wang, J. Lin and D. Li. 2006. Production of hydrolysate with antioxidative activity by enzymatic hydrolysis of extruded corn gluten. Appl. Microbiol. Biot. 73: 763-770.