廚餘是一種高濃度的有機廢棄物，其總 COD 約 280-430 g/L，VSS/SS 為 95% 以上，具有高濃度的固體物（約 210 g/L），且固體物COD電子分佈佔33.3%。本研究針對台南市的高澱粉濃度廚餘分別進行 20 次的採樣與特性分析，並估算廚餘的電子分布。固態碳水化合物及溶解態碳水化合物佔廚餘電子分布的最大宗，兩者的和約為總電子數的一半，而總有機氮所佔的比例則相對較小，為 17.3%。揮發酸約為8% 左右，而揮發酸中，又以乳酸的濃度最高，為5 g/L。本研究以一實驗室規模的 3 L 廚餘厭氧氫醱酵槽來進行其高澱粉濃度廚餘水解、酸化、產氫之程序研究，反應槽的形式定義為「間歇性進流完全攪拌反應槽」（Intermittent - Continuous Stirred Tank Reactor, I-CSTR）。在經過連續 250 天的長期操作，於第3-2試程發現有最高的平均體積產氫速率為 2.2 L-H2/L/day，該試程操作在水力停留時間為八天，間歇性進流頻率為24小時一次。最高的產氫 yield 也在第3-2試程，為 2.1 mmole-H2/g-COD；揮發性固體物降解率，在第2試程有最佳表現為47%，此試程操作在水力停留時間四天，間歇性進流頻率為12小時一次。在澱粉水解酶的部分，則是在第1試程有最大活性11 U/mL，而在第3-2試程有最佳固體碳水化合物水解為45%。然而，由時間序列採樣分析實驗，得知各試程中水解酶是足夠將固態碳水化合物完全水解的，且最大理論還原糖生成量為176 g-glucose eq./L/hr；在試程2中可有最大一階懸浮性固體物降解常數，值為0.04 hr-1。由降解米飯的產氫潛能生物活性測試，得知試程2有較佳的比產氫速率為0.15 mmole-H2/g-VSS/hr。由酵素動力模式及水解酶最佳化測試，可得KM值為34 g/L，最大速率（Vmax）為3.6 U/mL，且於溫度為55oC及使用磷酸緩衝溶液在pH 5.5或醋酸緩衝液在pH 4.4有最佳活性。以 I-CSTR 廚餘厭氧氫醱酵槽出流混合液進行 16S rRNA 基因選殖及 DNA 定序實驗，在挑選的 148 個 clones 中發現 27 個 OTUs（operational taxonomic units）。42% 的菌比對最接近為 Thermoanaerobacterium thermosaccharolyticum（相似度為 98%），另有比對到Clostridium sp.（相似度為95%，約佔 24%）以及兩株乳酸菌，分別為Lactobacillus panis (相似度99%, 約佔16%)及Lactobacillus amylovorus（相似度99%, 約佔10%）。其中，Thermoanaerobacterium thermosaccharolytium及Lactobacillus amylovorus被報導為具有分泌澱粉水解酶的能力。在 I-CSTR 廚餘厭氧氫醱酵槽 14 次的尾端修飾限制片段長度多形性（terminal restriction fragment length polymorphism, T-RFLP）圖譜分析中，發現反應槽內的微生物在不同操作條件下，其微生物菌相的組成皆已主要的4 OTUs為主，其中TKW-HPB-2（Thermoanaerobacterium thermosaccharolytium）在三試程皆佔有30%以上，而TKW-HPB-1（Clostridium sp.）則會在試程穩定後呈為優勢菌。

Kitchen waste was a kind of waste that contains highly nutritive organic compounds (total COD was about 280-430 g/L and VSS/SS ratio was higher than 95%). The total solid fraction was about 200 g/L and occupied 33% of total COD. The electron distribution of Tainan City kitchen waste was calculated according to the analyses data (n=20). Solid and soluble carbohydrate were the most dominant items of all which occupied about 50% of electron while total protein only about 17.3%. Electron distribution of VFAs (volatile fatty acids) was about 8% which were mainly contributed by 5 g/L of lactic acid. A 3 L bench scale of bio-hydrogenation I-CSTR (Intermittent - Continuous Stirred Tank Reactor) was established for kitchen waste treatment and bio-energy recovering process by anaerobic operation. Within 250 days of continuously long-term operation, the maximum averaged hydrogen production rate of 2.2 L H2/L-day was observed in run 3-2 with 8-day hydraulic retention time (HRT) and 24-hour intermittent feeding period. The highest average hydrogen yield of 2.1 mmol-H2/g-COD was also observed in run 3-2. However, the best VSS removal of 47% was occurred in run 2 which was operated at 4-day HRT and 12-hour intermittent feeding period. According to the analyses of amylase and reducing sugar, the maximum average amylase activity was about 11 U/mL in run 1, but the maximum solid carbohydrate hydrolysis rate was about 45% in run 3. To clarify the starch hydrolysis mechanism, amylase activity of effluent and time series profile between each operated period were monitored, and the white rice batch test was also studied. According to the results of time series profile analyses, amylase activity was sufficient for starch hydrolysis and the maximum theoretical reducing sugar production of 176 g-glucose eq./L was calculated. According to the results, the maximum first order constant of VSS degradation of 0.04 hr-1 was occurred in run 2. At the white rice batch test, the hydrogen producing bacteria taken from run 2 achieved better hydrogen production rate of 264 mL-H2/hr than the other ones. According to the enzyme kinetic study, the KM was 17 g/L and the maximum amylase activity was 1.5 U/mL. The environmental factors effects of amylase in the supernatant were also studied. The best reacted temperature was found to be 55oC. With phosphate buffer, the best amylase activity achieved when pH was at 5.5; with acetate buffer, the best one was observed at pH = 4.4. As the results of 16S rDNA-based cloning screening of the anaerobic microbes taken from I-CSTR bio-hydrogen reactor, the 27 operational taxonomic units (OTUs) were found in 148 clones. Among these clones, 42% of clones could be identified as Thermoanaerobacterium thermosaccharolyticum (98% similarity) and 24% of clones were identified as Clostridium sp. (95% similarity). The other two dominant clones were identified as Lactobacillus panis (99% similarity, abundance of 16%) and Lactobacillus amylovorus (99% similarity, abundance of 10%). Four dominant OTUs were observed during different runs by 14 times of T-RFLP analyses. Thermoanaerobacterium thermosaccharolyticum always exists in the system and amounted to 30%. However, Clostridium sp. would become dominant when the system was steady operated.

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