褐藻多醣(Fucoidan)是一種富含岩藻糖(Fucose)的硫酸化多醣，主要存在於褐藻中。許多研究指出褐藻多醣具有多種生物活性如調節免疫力，抗腫瘤，抗氧化，抗病毒且能抑制血管生成阻止癌細胞擴散，故頗具商業化開發之潛力。因此，尋找高效率褐藻多醣生產途徑，以增加產能、降低生產成本，是值得研究開發的課題。
本研究首先篩選出三種本土褐藻，分別是莢托馬尾藻(Sargassum siliquosum)、半葉馬尾藻(Sargassum hemiphyllum)以及匍枝馬尾藻(Sargassum polycystum)，利用傳統熱水萃取法對三種馬尾藻進行褐藻多醣的萃取，其中，莢托馬尾藻可得到最高的多醣產率，且岩藻糖的比例也較其他兩株馬尾藻高，結果顯示多醣含量佔莢托馬尾藻乾重的5.08%，而萃取出來的多醣中有46.5%為岩藻糖。接著，為了能再提高多醣的產率，分別對超聲波輔助萃取法以及微波輔助萃取法進行測試與比較。實驗結果發現，超聲波輔助萃取法對莢托馬尾藻褐藻多醣的萃取效果並不好，超聲波會導致多醣的降解而降低產率，因此不適合用作褐藻多醣的生產。而微波輔助萃取法具有較高的效率，在750 W, 10分鐘及固液比15 mL/g的條件下，多醣產率提高到6.94%。完成萃取條件的最適化之後，為了提高褐藻多醣的純度，本研究對幾種純化策略進行了測試，經過等電點沉澱及氯化鈣沉澱後，萃取物中大部分的蛋白質及醣醛酸成功地被移除，接著用陰離子層析管柱吸附褐藻多醣，分離其他副產物，再利用2 M的氯化鈉溶液以梯度方式進行洗脫，得到純化的馬尾藻萃取物，而多醣的含量從33.31%提高至78.26%且有70.94%的多醣回收率。
第二部分為褐藻多醣生物活性的分析，本研究分別對莢托馬尾藻萃取物及純化後的褐藻多醣的抗氧化、抑制脂質合成以及抗發炎能力進行測定。結果顯示莢托馬尾藻萃取物具有很高的抗氧化能力，但不具有抑制脂質合成及抗發炎的效果，而純化過後的褐藻多醣其抗氧化能力降低，然而被證實可以抑制細胞中28.9%的脂質合成及14.8%促炎因子(TNF)-α的產生。
雖然從莢托馬尾藻中萃取出的褐藻多醣具有抗氧化、抑制脂質合成以及抗發炎的能力，但為了能進一步提升其活性，本研究使用過氧化氫對多醣進行部分水解，探討分子量大小對褐藻多醣生物活性的影響。而從結果發現抗氧化能力會隨其分子量下降而提高，當分子量由107.3 kDa降低至3.2 kDa時，褐藻多醣清除DPPH自由基的半效應濃度(EC50)由2.58 mg/mL降低至1.82 mg/mL。此外，褐藻多醣在分子量30-100 kDa的區間中表現出相似的脂質抑制及抗發炎能力，然而當分子量下降至3 kDa時，效果大幅度提升，可抑制71.1%的脂質合成和36.7%促炎因子的產生。接著利用SO3-DMF法提高小分子褐藻寡醣的硫酸根含量，探討硫酸根含量對褐藻寡醣生物活性的影響。結果顯示當硫酸根含量由18.7%提高至32.1%，DPPH的EC50由1.82 mg/mL降低至0.86 mg/mL，且抗發炎活性也有效提高，然而脂質抑制能力卻由71.1%降低至5.3%。
最後，本研究利用ESI-MS來分析莢托馬尾藻中褐藻多醣的結構，由圖譜顯示，褐藻多醣的主要結構為L-岩藻糖以α-(1,3)-糖苷鍵組成，且具有許多硫酸基在2號碳及4號碳的位置上，多醣上有許多分枝，主要為D-半乳糖以(1,3)-糖苷鍵組成，分枝點位於岩藻糖的4號碳上，而半乳糖上的硫酸基主要位於4號及6號碳上，也有少數分布在2號碳。

Fucoidan, a group of fucose-rich sulfated polysaccharides found in brown algae, has been shown to possess a variety of human health-related biological activities, such as immune competence regulation, antitumor, antioxidant, antivirus, and inhibition of angiogenesis. Therefore, fucoidan has a high potential for commercialization. In light of this, it is worthwhile to identify more efficient production strategies to improve the productivity of fucoidan and reduce its production cost.
In this study, three brown algae species identified as Sargassum siliquosum, Sargassum hemiphyllum, and Sargassum polycystum were examined for their potential for fucoidan production. First, conventional hot water extraction method was applied to extract fucoidan from the algal biomass. Among them, S. siliguosum showed the highest fucoidan production and fucose content, giving a total sugar yield of 5.08% with 46.5% of fucose in total sugar. To increase fucoidan yield, ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) were evaluated. The results revealed that UAE causes polysaccharide degradation and decreases fucoidan yield. In contrast, MAE showed high efficiency in fucoidan production, which gave a total sugar yield of 6.94% dry weight under the condition of 750 W, 10 minutes, and a liquid/solid ratio of 15 ml/g. The crude fucoidan was then purified to gain high purity. With the isoelectric focusing treatment and the addition of calcium chloride to final concentration of 20 g/L, most of the protein and alginate were removed. Anion-exchange chromatography (AEC) was subsequently carried out for further purification. Fucoidan was bound on the resin and eluted with 2 M sodium chloride in a linearly increasing gradient. The fucoidan content finally increased from 33.31% to 78.26% with 70.94% of total sugar recovery.
This study also assayed the antioxidant, anti-lipogenesis, and anti-inflammatory activity of the extracted fucoidan, which were investigated by DPPH scavenging effect, lipid synthesis inhibition, and the inhibition of pro-inflammatory cytokine (TNF)-α production, respectively. The results indicated that crude extract of fucoidan showed a high DPPH scavenging effect, with a EC50 of 0.34 mg/mL, while the inhibition of lipid synthesis and TNF-α production were not determined. After purification, DPPH scavenging effect decreased, but anti-lipogenesis activity and anti-inflammatory activity were discovered. The results show that the purified fucoidan product could inhibit 28.9% of lipid synthesis and 14.8% of TNF-α production.
To improve the potential biological activity of the fucoidan products, partial hydrolysis was carried out to gain lower molecular weight compounds. With hydrogen peroxide treatment, fucoidan fractions with different average molecular weight (Mw) were obtained. The results showed that antioxidant activities increased with decreasing Mw and the EC50 dropped from 2.58 to 1.82 mg/mL when the Mw decreased from 107.3 to 3.2 kDa. In addition, anti-lipogenesis and anti-inflammatory significantly enhanced when the Mw decreased to about 3 kDa, giving a reduced level of 71.1% and 36.7%, respectively. Then, low molecular weight fucoidan was treated with SO3-DMF method to gain high sulfate content. The results indicated that when sulfate content increased from 18.7% to 32.1%, EC50 of DPPH decreased from 1.82 mg/mL to 0.86 mg/mL, and the anti-inflammatory activity also increased; however, the anti-lipogenesis activity decreased.
In the last part of this study, ESI-MS was used to analyze the structure of fucoidan obtained from S. siliquosum. The ESI-CID-MS/MS spectra indicated that fucoidan has a backbone composed of (1-3)-linked L-fucose residues, and the sulfate groups are located at C-2 and C-4 of fucose residues. Branches are galactose residues with (1,3)-linkages and the branching point is at C-4 of fucose residues. The sulfate groups on galactose was mainly presented at C-4 and C-6 of galactose residues and some located at C-2.

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