由於微藻具有油脂含量高、生長快速、佔地面積小與淡水需求度低等特性，因此對於生質柴油等可再生能源而言，微藻是一種極具潛力的料源。然而，由於微藻養殖所費不貲，從稀薄的藻液中收集這些微小的微藻細胞(<30 µm)時有著許多不確定性，因此微藻生質柴油目前仍未商業化。本研究以淡水小球藻Chlorella vulgaris ESP-3 1、海水小球藻Chlorella sp. 與海水衣藻Chlamydomonas orbicularis Tai-04等三種微藻進行以甲殼素與聚氯化鋁(PACl)為絮凝劑之收藻方法測試。而為了能在最少的絮凝劑使用量下達成 90%以上的藻體收集率，各種會影響藻體絮凝的因素如絮凝劑劑量、藻液pH值、攪拌轉速與時間、藻液中的生物質濃度以及藻體內的油脂含量皆是測試之項目。研究結果顯示，在最佳的操作條件下，Chlorella vulgaris ESP-31收集之最佳絮凝劑劑量分別為0.013 g甲殼素/g 藻體與0.101 g PACl/g 藻體。對海水小球藻Chlorella sp. 而言，使用甲殼素為絮凝劑之最佳劑量為0.044 g/g 藻體。衣藻的收集在以不同氮濃度培養(1N, 2N and 3N)下，PACl的最佳劑量分別為0.491、0.175與0.491 g/g。進一步以所收集的Chlorella vulgaris ESP-31做為原料，進行以固定化脂解酵素為觸媒之直接轉酯化程序以合成生質柴油。研究結果顯示，當以PACl絮凝收集到的ESP-31為原料時，酵素活性明顯地被抑制，因此生質柴油產率極低。然而當以甲殼素絮凝收集到的ESP-31為料源時，固定化脂解酵素則能正常運作，且其所獲得之生質柴油產率高達97.6 wt%，這結果與以離心收集到的藻體之生質柴油產率相近(97.1 wt%)。此外，此固定化脂解酵素連續重複使用四次，其活性並沒有明顯地減少。此結果顯示，以甲殼素輔助絮凝小球藻Chlorella vulgaris ESP-31所獲得之藻體不需要再經過其他的前處理，即可直接進行酵素之直接轉酯化生產生質柴油。

Microalgae has been considering as a potential feedstock for renewable energy production, especially for biodiesel, due to their high oil content, fast growth rate, low land and freshwater requirements. However, industrial application of microalgal biodiesel has been hindered due to the high production costs and the uncertainties during biomass harvesting.
In this study, three microalgal strains namely Chlorella vulgaris ESP-31 (fresh water), Chlorella sp. (sea water) and Chlamydomonas orbicularis Tai-04 (sea water) were harvested by coagulation method using chitosan and polyaluminium chloride (PACl) as coagulant agents. The effect of parameters influencing harvesting efficiency, such as coagulant dose, pH, mixing rate, mixing time, biomass concentration, and lipid content, were examined. The results show that under the optimal conditions, the amount of chitosan and PACl needed for harvesting 1 g of dry microalgal biomass C. vulgaris ESP-31 were 0.013 g and 0.101 g, respectively. For Chlorella sp., 0.044 g chitosan/g micoalgae biomass was required. For C. orbicularis Tai-04 grown under different nitrogen source concentration (1N, 2N, and 3N), the required PACl dose was 0.491, 0.175 and 0.491 g PACl per gram of Tai-04. The harvested C. vulgaris ESP-31 was directly employed as the oil source for biodiesel production via enzymatic-transesterification catalyzed by immobilized lipase (Burkholderia sp.), and chemical transesterification catalyzed by synthesized solid catalyst (SrO/SiO2). Both enzymatic- and chemical-transesterification were significantly inhibited in the presence of PACl, while the immobilized lipase worked well with wet chitosan-coagulated ESP-31. The conversion of biodiesel by immobilized lipase from chitosan-coagulated ESP-31 reached 97.6 w/w (oil), which is at a level comparable to that of biodiesel conversion from centrifugation-harvested microalgae (97.1% w/w oil). The solid catalyst SrO/SiO2 worked well with water-removed centrifuged ESP-31, and biodiesel conversion reached around 80% w/w (oil). However, water-removed chitosan-flocculated ESP-31 only slightly induced SrO/SiO2 in transesterification with methanol, and thus the biodiesel conversion only reached 55.7–61.4% w/w (oil). The biocatalyst seems to have higher potential than solid catalyst SrO/SiO2, since the former can be applied with wet microalgae, thus eliminating the need to dry the microalgae biomass before the transesterification reaction. Moreover, the biocatalyst can be easily separated and re-used for up to three cycles in repeated-batch reactor without significantly decreasing its activity.

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