台灣現行之廚餘回收每天可達約1,900噸，是來源豐富的有機廢棄物之一，其中廚餘之70%以上為水，其液體部分可夾代豐富之營養成分適合用於產氫。本研究主要產氫基質選用廚餘過篩液，是將廚餘滲出的液體部分經過1~2 mm篩孔過篩，可有效控制懸浮固體物大小及濃度在26 g-SS/L(VSS/SS=0.91)，其目的是降低SS含量使流體化床能操作順利，而流體化床的優點為藉由流力控制使懸浮固體物(基質及菌體)有更長停留時間且濃度更高的在反應區中反應。而此廚餘過篩液也能提供高濃度COD約104 g/L及總醣濃度總26.5 g /L供微生物產氫利用，其中碳水化合物的電子含量最高，約佔總COD之30.3%(溶解性佔總COD之26.9%)，其次為油脂佔23%、蛋白質佔22.7%，再來為乳酸佔14.9%及乙酸2.6%。另外其TVS約73 g /L，可知以溶解性有機物為多。從批次實驗也顯示，廚餘過篩液本身所夾帶之嗜酸性微生物可有效分解其內養分且具良好產氫能力。本研究還有額外添加狼尾草當輔助基質與廚餘過篩液混合，其目的有三個：(1) 發揮流體化床擔體功能；(2)直接提高基質之有機負荷；(3) 培養出分解纖維及半纖維素之產氫菌。實驗所用的風乾狼尾草渣每克約可提供1.1 g-COD，有機成分佔總重的88.5%(含水率7.2%)，而總醣佔總重的51%，夾帶有總重7.5%的溶解性醣類、纖維素20%及半纖維素及其他醣類約佔23%。蛋白質佔8.5%。而本研究利用110 L厭氧流體化床(反應溫度55oC、pH=6、HRT=7.3 day及總迴流量35 L/min)以廚餘過篩液為主要基質和狼尾草為輔助基質在193天連續氫醱酵操作下，第三試程(廚餘過篩液+20 g/L狼尾草)在有機荷負為17.7 g-COD/L/day下，有最高產氫速率約為1.64 L-H2/L/day(比前面第一及第二試程提升約16.6%)，但是氫氣產率3.74 mmol-H2/g-CODin為各試程中最低的(也就是狼尾草中大部分醣類難分解而拉低產率)；第一試程(只進廚餘過篩液)在體積負荷為14.2 g-COD/L/day下，產氫速率約為1.41 L-H2/L/day，其中氫氣產率約4.0 mmol-H2/g-CODin為各試程中最高的。各試程溶解性醣類去除率均可達約92%。流體化床在各試程的操作中，出流主要代謝產物以丁酸產生最多(13~17 g/L) ，乙酸次之 (2.3~3.3 g/L) ，而丙酸(生成345 mg/L)及乙醇(生成400 mg/L)生成量相當少，顯示此槽內微生物與廚餘過篩液在於產氫代謝是相當旺盛的，不利於產氫的代謝途徑並不顯著。另外值得注意的是各試程進料廚餘過篩液中高濃度乳酸(10.7~14.5 g/L)有被顯著降解的現象，其中第二試程有最高乳酸降解率，平均98.3%。由本研究乳酸降解試驗可證實槽內微生物可利用乳酸加上少量乙酸共基質代謝生成氫氣及丁酸。故可將槽內主要產氫途徑分為:
(1)乳酸代謝產氫
Lactate + 0.4 HAc +0.7 H+ → 0.7 HBu + 0.6 H2 + CO2 + 0.4 H2O
(2)醣類代謝產氫
C6H12O6 + 2H2O → 4H2 + 2CO2 +2HAc
C6H12O6 → 2H2 + 2CO2 + HBu
可知乳酸代謝產氫時，醣類代謝產氫所產生的乙酸會被消耗掉，所觀測到的乙酸生成濃度會減少，而乳酸代謝產氫也會同時生成丁酸，會與醣類代謝產氫所產生的丁酸累加，所觀測到的丁酸濃度會大量增加。以乳酸降解率最好的第二試程(98.3%)及第三試程(94.9%)為例，實測值的丁酸產生量與乙酸產生量的莫耳比值HBu/HAc會相當高(Run 2: 5.19, Run 3: 3.64)，但將乳酸代謝干擾去除所得的醣類代謝之HBu/HAc會接近1(Run 2: 1.38, Run 3: 0.87) ，這表示槽內微生物利用醣類的反應接近此式: 3C6H12O6 + 2H2O → 8H2 + 6CO2 + 2 HAc + 2 HBu，故此式也顯示槽內最大的醣類代謝之氫氣產率約2.67 mol-H2/mol-hexose。而由揮發酸產量所推的理論產氫量與實際只誤差4.7~12.2%，顯示由其他非產氫代謝(如蛋白質)所產的乙酸及丁酸很少，而其中所降解的廚餘過篩液中之乳酸所產的氫氣約佔總產氫量的14.8%(Run 2)~17.1%(Run 3)，故對乳酸濃度相當高廚餘滲出液來說，本流體化床內的乳酸產氫菌群除了可大幅的增加其產氫潛能外，此乳酸代謝過程還會消耗酸度，這對連續流進料的產氫酸化系統之pH維持恆定操作有很大的幫助，可緩和醣類代謝產氫時的酸化程度而減少液鹼回饋量。另外槽內及廚餘中微生物經流體化床經100多天的狼尾草馴養，對纖維素(CMC)完全沒水解及產氫活性，但對對木聚醣(半纖維素)有良好水解產氫活性，其可能因狼尾草物料僅進行初步的物理破碎，纖維素的露出率有限，微生物較難接觸到(木質素及半纖維素包在纖維素外面)。故第三試程加最多狼尾草20 g/L有最好之產氫量，其多出的產氫量可能主要由狼尾草所夾帶的少量溶解性醣類(7.5%)及水解部分半纖維素所提供。在槽內微生物菌相探討方面，由SEM可觀測到廚餘及流體化床的微生物型態幾乎都以桿菌為主，另外由兩端帶螢光之T-RFLP前後端之分生檢測結果，與本研究團隊所做的廚餘產氫系統菌群clone之 T-RFLP資料庫做比對，本研究槽內主要之微生物可能為Clostridium sp.和Thermoanaerobacterium sp.(reverse: 307 bp & 317 bp)，這兩屬中很多菌種對多種複合醣類都具有產氫能力，而且皆為桿菌型態並有內生孢子能力。另外，乳酸降解試驗中產氫最好的組別在T-RFLP的forward 495 bp波峰，跟批次植種比有明顯的提升(4.3%升到36.1%)，故推測forward 495 bp位置可能為乳酸產氫菌，而在第二試程中乳酸去除率(98.3%)最好的時候，495 bp位置能成為主流波峰之一，最高可佔總族群的41.3%。然而在在Run 3中後期時，495 bp位置及其他較小的波峰位置幾乎快消失(只有最主流波峰不變)，可能是因為Run 3之進料SS過高，到中後期常會有堵塞問題，故在清理時會增加氧氣進入反應槽中的機會，使部分厭氧微生物可能會受到影響。

This study used kitchen waste sieved liquid (KWSL) (KW was sieved through 1~2 mm pore size) as a substrate for biohydrogenation by anaerobic fluidized bed reactor (AnFB). And later, different weights of napiergrass dregs were added to co-ferment with KWSL in long hydraulic retention time (HRT) (7.3 days).In Taiwan, the recycling amount of KW was up to 1900 ton/day in average. The moisture of KW was more than 75%, which (KWSL) could carry abundant nutrient and suit for biohydrogenation as well as the operation of AnFB (less suspended solid).The characteristic of KWML as follows: Total COD : 104,000 ± 16,000 mg/L (69% soluble) , Total Carbohydrate: 26,500 ± 6,300 mg/L (85% soluble ) , Total Org-N: 2,560 ± 270 mg/L (70% soluble) , lipid: 8,300 ± 3900 mg/L , suspended solid: 26,000 ± 5330 mg/L( 91 % volatile) , and it contained acetate (2,500 ± 340 mg/L) and high concentration lactate ( 10,700~14,500 mg/L ) . Napiergrass (1.1 g-COD/g- napiergrass dregs) is composed by about 20% cellulose and other lignin cellulose which are difficult to decomposition, this study expected that adding the napiergrass dregs with KWSL could degrade the cellulose and act as a kind of support material in the AnFBR.
The AnFBR has a draft tube in reacting zone, and let solid has longer retention time to reaction by controlling recycle flow rate .Working volume of the AnFBR is 110 L. The seeding was from the sludge which used KW vegetable as substrate to produce hydrogen. Operation parameters of the AnFBR: temperature:55oC , pH:6 ,HRT:7.3 days and recirculation liquid flow rate: 35 L/min
Total three runs: Run 1 (70 days) inputted KWSL only (organic loading rate: 14.2 ± 2.4 g-COD/L/day), and the hydrogen production rate (HPR) was 1.41 ± 0.27 L-H2/L/day. Run 2 (73 days) inputted KWSL with 5 g- napiergrass dregs/L , and the HPR was 1.41 ± 0.28 L-H2/L-day. Run 3 (47 days) increased the napiergrass concentration to 20 g- napiergrass dregs /L, and had the best HPR,1.64 ± 0.28 L-H2/L-day(Total biogas: 399.6 ± 40.4 L/L/day),in this study.
In metabolites aspect, the main volatile fatty acids (VFA) in the effluent were butyrate (13,000~17,000 mg/L) and acetate (2,300~33,00 mg/L) production. Especially, the high concentration lactate in KWSL was degraded obviously (maximum removal : 98.3% for Run 2),and from the batch test of lactate degradation, it showed that lactate was degraded and hydrogen was produced, which the reaction was similar to “Lactate + 0.4 Acetate +0.7 H+ → 0.7 Butyrate + 0.6 H2 + CO2 + 0.4 H2O”. Regarding KWSL contained high concentration lactate has advantages of biohydrogenation and reducing the feed-back of alkali.

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