近年來，以微藻作為生質能料源備受重視，因微藻生長快速且可經由光合作用固定二氧化碳並轉化生成生物量，而且某些微藻(如綠藻等)可將碳水化合物以澱粉與纖維素的型態大量累積儲存於藻體，因此被視為具有潛力作為生質酒精醱酵生產的料源之一。
本研究首先將台灣中南部水域所篩選出之四株本土微藻Chlorella vulgaris ESP-6、C. vulgaris FSP-E、C. vulgaris BS-C與Chlamydomonas orbicularis Tai-04進行微藻產糖之鑑定。結果顯示，在相同培養條件下C. vulgaris FSP-E可獲得最高的藻體生物量濃度及生產速率，分別為4.3 g/L 和0.18 g/L/d。這四株微藻中又以C. vulgaris FSP-E的碳水化合物含量及葡萄糖含量最高，分別約佔藻體乾重54.1% 和50.2%。此外，為了有效提升微藻生物量及碳水化合物產率，本研究探討不同氮源、不同光照強度、不同植藻量、不同氮源濃度、不同氮源缺乏天數對微藻體生物量及碳水化合物產率的影響。結果顯示，在1升的光生物反應器操作下，當光照強度為440 micromol m-2s-1、植藻量為0.14 g/L、氮源(尿素)濃度為0.56 g/L、氮源缺乏時間3天時，有最佳碳水化合物產率。最高碳水化合物與生物量生產速率分別為0.687 g/L/d 和1.363 g/L/d，其中碳水化合物含量及葡萄糖含量分別約佔藻體乾重的50.4±0.4%和46.9±0.4%。而在5升反應器以相同培養條件下氮源缺乏4天後，其最高碳水化合物與生物量生產速率分別為0.378 g/L/d 和0.792 g/L/d。
本研究接著將所獲得的藻體生物量進行水解糖化及酒精醱酵測試。在藻體酵素水解糖化之測試方面，當藻體濃度為20 g/L、藻體超音波振盪預處理為10分鐘、水解酵素用量為內切型纖維素分解酵素0.65 U/mL、beta-葡萄糖苷酶0.30 U/mL、澱粉酶0.75 U/mL時，有最佳水解糖化效能，其最佳葡萄糖產率為90.4%。接著，以兩階段水解糖化醱酵程序（SHF）在此酵素水解條件下進行酒精生產之探討。結果發現，以20 g/L藻體為碳源時，在SHF系統下可達酒精產量及酒精理論產率分別為3.55 g/L及80.1%。在硫酸水解糖化之測試方面，當藻體濃度為60 g/L、1%硫酸濃度下，有最佳水解效能，最佳葡萄糖產率為93.6%。並以兩階段水解糖化醱酵程序（SHF）在此硫酸水解條件下進行酒精生產之探討。結果發現，以藻體（60 g/L）為碳源時，在SHF系統下可達酒精產量及酒精理論產率分別為11.7 g/L及87.6%。由上述結果顯示，本研究利用各種策略確實能有效提升微藻生物量及碳水化合物產率，並可以有效地利用兩階段水解醱酵策略轉化藻體醣類以產生質酒精。

Microalgae are considered as one of the most promising renewable feedstock for bioethanol production due to the advantages of fast growing, efficient carbon dioxide fixation, and potentially containing high carbohydrate contents (mainly in the form of cellulose and starch). Therefore, this study aimed to evaluate the potential of carbohydrate production from microalgae for bioethanol production.
First, four indigenous microalgal strains, namely Chlorella vulgaris ESP-6, C. vulgaris FSP-E, C. vulgaris BS-C, and Chlamydomonas orbicularis Tai-04, were investigated to identify their growth rates and carbohydrate content. Among the four strains examined, C. vulgaris FSP-E showed the highest biomass concentration and carbohydrate productivity, which was 4.3 g/L and 0.18 g/L/d, respectively. C. vulgaris FSP-E also showed the highest carbohydrate and glucose content, which was 54.1% and 50.2%, respectively. Therefore, the FSP-E strain was selected as the target microalga strain in this work.
The biomass and carbohydrate productivity of C. vulgaris FSP-E were further improved by adjusting the light intensity, inoculum size, nitrogen type and concentration, and nitrogen starvation period. The optimal conditions for carbohydrate productivity occurred when the microalga culture was controlled at light intensity, 440 micromol m-2s-1; inoculum size, 0.14 g/L; nitrogen source, urea; urea concentration, 0.56 g/L; nitrogen starvation period, 3 days in a 1L glass-vessel type photobioreactor (PBR). The carbohydrate productivity, biomass productivity, carbohydrate content, and glucose content were 0.687 g/L/d, 1.363 g/L/d, 50.4±0.4%, and 46.9±0.4%, respectively. When this cultivation condition was applied in 5L PBR, the carbohydrate and biomass productivity was lower at 0.378 g/L/d and 0.792 g/L/d, respectively.
Enzymatic hydrolysis of the microalgae-based carbohydrate was the most efficient when 20 g/L of microalgal biomass was first pretreated by 10 min sonication and hydrolyzed by a cocktail enzyme mixture consisting of endoglucanase: 0.65 U/mL, beta-glucosidase: 0.30 U/mL, amylase: 0.75 U/mL. The highest glucose yield obtained from the foregoing procedures was 90.4%. The enzymatic hydrolysate of C. vulgaris FSP-E (glucose content =7.58 g/L) was used for ethanol production via SHF process, producing 3.55 g/L ethanol, which represents 80.1% of the theoretically maximum yield. As for acidic hydrolysis of the microalga biomass, the highest glucose yield (93.6%) was achieved when microalgal biomass concentration was 60 g/L and sulfuric acid concentration was 1%. The acidically hydrolyzed C. vulgaris FSP-E biomass was also used for ethanol production by SHF process, giving rise to an ethanol concentration of 11.7 g/L and a yield of 87.6% of the theoretical yield. These results indicate the feasibility of producing carbohydrate from microalgal biomass for fermentative bioethanol production.

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