臺灣博碩士論文加值系統

食品發酵為最古老的食品加工及防腐方法之一。一般而言，在泰國利用環境中自
然存在之微生物來製作 Isan 香腸需要 3-5 天之發酵時間，近年來消費者逐漸重視食品衛生，並且極度要求安全且健康之產品。發酵肉製品之工業製程中通常會添加糖類來促進發酵進行。在發酵與熟成 (ripening) 期間，乳酸菌 (lactic acid bacteria, LAB)會將糖類及其他碳水化合物轉變成乳酸，此舉可降低產品之 pH 值達到防腐之效果，並可產生發酵產品之特殊感官特性。由於糖類獨特之特性，使得不同糖類在發酵過程中被微生物利用之方式可能不盡相同。因此，本研究旨在探討不同糖類添加量 (蔗糖及葡萄糖) 及接種乳酸菌菌酛 (Lactobacillus plantarum starter culture) 對 Isan 香腸理化、微生物、質地及感官特性之影響。
試驗一結果顯示，在發酵 4 天內，添加菌酛之香腸 (3SLp 與 12SLp)其 pH 值較未添加菌酛之香腸 (3S 與 12S) 低，顯示添加菌酛可加快發酵過程，而不同蔗糖添加量之香腸 (3SLp vs. 12SLp 與 3S vs. 12S ) 間無顯著差異。各組間水活性及外觀顏色 (L*,a*及 b*值) 雖有顯著差異，但差異較小。相較於未接種之香腸，接種菌酛之香腸其硫巴比妥酸反應物 (thiobarbituric acid reactive substances, TBARS) 及揮發性鹽基態氮(volatile basic nitrogen, VBN) 數值均較低。在發酵期間各組之總生菌數及乳酸菌數迅速地上升，之後呈現下降趨勢。在質地及感官特性方面，接種菌酛之香腸有較高之硬度，品評員則較喜歡添加 1.2 %蔗糖且接種菌酛之香腸。
試驗二結果顯示，在發酵及貯存期間，相較於同時添加 0.6 %葡萄糖與 0.6 %蔗
糖或添加 1.2 %蔗糖 (12S) 之香腸，添加 1.2 %葡萄糖 (12G) 可以顯著降低香腸之 pH值。在發酵及貯存期間各組之 L*值及 b*值均顯著上升，a*值則顯著下降，且 12G 之L*值顯著高於 12S。各組之總生菌數及乳酸菌數在發酵期間持續上升，隨後在貯存期間內持續下降，相較於 6S6G 與 12S，12G 有較高之乳酸菌數及總生菌數。在發酵及貯存期間，TBARS 與 VBN 數值持續上升，且 12G 之 TBARS 與 VBN 數值顯著低於各組。在發酵及貯存期間，硬度及咀嚼性持續上升，而彈性及凝聚性則呈現先升後降之趨勢。在貯存期間，相較於 6S6G，12G 有較高之硬度、咀嚼性及較低之彈性。雖然各組間感官特性均無顯著差異，但 12G 在所有品評項目上均較其他組高。由試驗二結果可知，在已接種菌酛之 Isan 香腸製程中添加 1.2 %葡萄糖可以得到較佳之產品品質。綜觀上述，添加蔗糖或葡萄糖且配合接種 107 CFU/g 濃度之 L. plantarum 菌酛不僅可以增加 Isan 香腸之理化、微生物及質地特性品質，更可增加感官品評接受度及保存期限。

Food fermentation is one of the oldest food processing and preservation methods.
Generally, in Thailand it takes 3–5 days to ferment Isan sausages with adventitious
microorganisms which present naturally. Nowadays, consumers are more concerned about the food safety and highly demand for safe and healthy products. Sugars are usually included for the industrial manufacture of fermented meat products. During fermentation and ripening, lactic acid bacteria (LAB) convert glucose and other carbohydrates to lactic acid, thus further decrease pH, preserve products, and contribute to the development of the typical organoleptic characteristics of fermented products. Various sugars might have been utilized differently in fermentation due to their unique characteristics. Therefore, the objective of this study was to investigate the effects of adding different amounts of sugars (sucrose and glucose) with inoculation of the starter culture (Lactobacillus plantarum) on
physicochemical, microbiological, texture, and sensory properties of Isan sausages.

In experiment 1, the results showed that the sausages that inoculated with starter cultures (3SLp and 12SLp) fastened the fermentation process than the ones without inoculation (3S and 12S) in accordance to the lower pH in 4 days, whereas there was no significant difference in pH values between the samples with different sucrose adding levels (3SLp vs. 12SLp and 3S vs. 12S). Less difference was observed in the water activity and L*, a*, and b* values between samples. Inoculated samples had significantly lower thiobarbituric acid reactive substances (TBARS) and volatile basic nitrogen (VBN) values than the ones without inoculation. The total viable bacteria and lactic acid bacteria counts increased rapidly during the fermentation and then decreased. Inoculated samples had significantly higher total plate counts and lactic acid bacteria counts than the ones without inoculation. For the texture profile, inoculated samples had higher hardness and panels preferred the sensory characteristics of the inoculated and 0.3% sucrose-added sausages.

In experiment 2, the result showed that samples with addition of 1.2% glucose (12G) had a significantly lower pH value as compared to samples with addition of 0.6% glucose plus 0.6% sucrose (6S6G) and 1.2% sucrose (12S) during process. The L* values (lightness) and b*values (yellowness) of samples increased and a* values (redness) decreased during process. The 12G samples had significantly higher L* values than the 12S samples during storage. The lactic acid bacteria counts and total plate counts of samples increased during fermentation and then slightly decreased during storage. The 12G had a significantly higher lactic acid bacteria counts and total plate counts than the 6S6G and 12S samples. The TBARS and VBN values increased during process. The 12G samples had significantly lower TBARS and VBN values. Hardness and chewiness increased during process while the springiness and cohesiveness were increased and then decreased during process. During storage, the 12G samples had higher hardness and chewiness, and lower springiness than the 6S6G samples during storage (P < 0.05). There was no significant difference on the sensory characteristics between treatments (P > 0.05). In sausages with adding 1.2% glucose into formula during manufacturing led to an inoculated fermented Isan sausage with superior qualities.

In conclusion, adding sucrose or glucose combined with inoculation of L. plantarum at level of 107 CFU/g not only enhances the physicochemical, microbiological, and textural qualities of the Isan sausages, but also increases the sensory acceptance and shelf-life of products.

Chapter 1. Introduction ............................... 1
Chapter 2. Literature review ................................................................... 3
2.1. Fermentation .................................................................................... 3
2.2. Isan sausage ........................................................................................ 4
2.2.1. Customary material of Isan sausage ........................................................... 5
2.2.2. Starter culture for fermentation sausages ........................................................... 9
2.3. Inoculation of starter culture ..................................................................... 9
2.3.1. Lactic acid bacteria ......................................................................... 9
2.3.2. Lactic acid bacteria as functional starter cultures ................................. 10
2.4. Addition of sugars for fermentation ............................................................. 15
2.4.1. Sucrose ..................................................... 16
2.4.2. Glucose ...................................... 16
CHAPTER 3: Experiment 1 ..................................................... 18
3.1. Abstract ..................................................... 18
3.2. Introduction .............................................. 19
3.3. Materials and methods ..................................................................... 20
3.3.1. Preparation of starter culture .............................................................................. 20
3.3.2. Preparation of Isan sausage ................................................................................ 20
3.3.3. Physicochemical analysis and instrumental color evaluation ........................ 21
3.3.4. Microbiological analysis ..................................................................................... 21
3.3.5. Proximate Analysis .............................................................................................. 22
3.3.6. Textural and sensory evaluation ........................................................................ 22
3.3.7. Statistical analysis ................................................................................................ 23
3.4. Results and discussion............................................................................................ 23
3.4.1. pH and titratable acidity ...................................................................................... 23
3.4.2. Water activity (aw) ............................................................................................... 24
3.4.3. Color changes ....................................................................................................... 25
3.4.4. Microbial quality .................................................................................................. 26
3.4.5. Thiobarbituric acid reactive substances (TBARS) and volatile basic nitrogen (VBN) .................................................................................................... 27
3.4.6. Proximate composition and weight loss .............................................. 28
3.4.7. Texture profile analysis ....................................................................................... 29
3.4.8. Sensory evaluation ............................................................................................... 30
3.5. Conclusion.............................................................................. 32
3.6. References ............................................................................................ 32
Chapter 4. Experiment 2 ....................................................................................... 51
4.1. Abstract .............................................................................. 51
4.2. Introduction .................................................................................. 52
4.3. Materials and Methods ...................................................................... 53
4.3.1. Preparation of starter culture ................................................................ 53
4.3.2. Preparation of Isan sausage ................................................................. 53
4.3.3. Physicochemical analysis and instrumental color evaluation ........................ 54
4.3.4. Microbiological analysis ..................................................................................... 54
4.3.5. Textural and sensory evaluation .................................................................. 55
4.3.6. Statistical analysis ........................................................................... 55
4.4. Results and Discussion .................................................................... 55
4.4.1. pH, titratable acidity, and water activity .................................................. 55
4.4.2. Color changes ............................................................................................ 57
4.4.3. Microbial quality .............................. 58
4.4.4. Thiobarbituric acid reactive substances (TBARS) and volatile basic nitrogen (VBN) ...................................................................................... 58
4.4.5. Texture profile analysis ...................................................................... 59
4.4.6. Sensory evaluation ................................................................................ 60
4.5. Conclusion............................................ 61
4.6. References ...................................................... 61
Chapter 5. Conclusion ...................................................... 74
REFERENCES ...................................................... 75
APPENDICE............................................. 87

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