農業部-水產試驗所全球資訊網農業部-水產試驗所全球資訊網

藻已被研究出具有多種對人體有益的活性成分，包括活性胜肽或酚類化合物，本研 究為利紅葡萄藻(Botryocladia leptopoda)多元化利用，以紅葡萄藻蛋白質經酵素 水解後之水解物，進一步以體外試驗探討其蛋白質水解物之生理活性，藻渣經乾燥 後，進一步萃取多酚。紅葡萄藻經酵素及鹼處理萃取蛋白質後，利用蛋白質鑑定技 術鑑定蛋白質種類，並從其中挑選兩個蛋白質進行後續潛在活性胜肽探討，挑選依 據為蛋白質來源之親緣關係與紅葡萄藻較為相近者且序列覆蓋率較高者，結果以核 酮糖-1,5-二磷酸羧化酶/加氧酶及藻紅蛋白β鏈最為合適。2種蛋白質序列以 BIOPEP-UWM活性胜肽資料庫進行分析，結果顯示紅葡萄藻蛋白質具有ACE抑制活性、 DPP-4抑制活性及抗氧化活性。紅葡萄藻蛋白質以六種不同酵素進行水解，並測試各 酵素之最適水解條件、ACE抑制活性及DPP-4抑制活性，結果顯示較適酵素水解之紅 葡萄藻蛋白質水解物具有最強的ACE抑制活性以及DPP-4抑制活性，與BIOPEP-UWM模 擬結果相符。另以分子量進行區分(<3 kDa、3-10 kDa、>10 kDa)，結果以>10 kDa之抑制活性最佳，接著進一部以細胞實驗證實其調節血糖之功效，結果顯示濃度 3000μg/ml以上的組別表現出細胞毒性，其餘以下濃度的組別及不同濃度之市售藥 品皆不具細胞毒性，因此後續抑制實驗僅針對2000 μg/ml以下的組別進行探討，結 果發現濃度2000 μg/ml 約可抑制3成的DPP-4活性，且具劑量依賴性。另將藻渣添 加乙醇萃取多酚，結果每克乾藻可萃取215mg多酚產物，顯示：紅葡萄藻萃取機能性 胜肽後，再以乙醇提取多酚為較適化萃取條件。綜合上述結果，紅葡萄藻活性胜肽 具有開發為血壓及血糖保健產品之潛力，另可探討多酚之功效作用。

A variety of active ingredients is have been developed from algae, including active peptides or phenolic compounds.  This study is a diversified applied of Botryocladia leptopoda. We hydrolyzes the algae protein by enzyme hydrolysis and use in vitro experement to investigated the physiological activity of the protein hydrolyzate. The dried algae residue is further to extract polyphenol. After extracting the protein by enzyme and alkali treatment, the algae protein uses protein identification technology to identify the protein species, and selects two proteins for subsequent potential active peptides. The selection is based on the genetic relationship and sequence coverage. The ribulose1,5-bisphosphate carboxylase/oxygenase and phycoerythrin βchain are most suitable. The two protein sequences were analyzed by the BIOPEP-UWM database, and the results showed that the algae protein has ACE inhibitory activity, DPP-4 inhibitory activity and antioxidant activity. The algae protein was hydrolyzed by six different enzymes, and the optimum hydrolysis conditions of each enzyme were tested. It also - 1 - 1082420 explore the ACE inhibitory activity and DPP-4 inhibitory activity of each protein hydrolysate. The results showed that the protein hydrolysate had the strongest ACE inhibitory activity and DPP-4 inhibitory activity, consistent with the BIOPEP-UWM simulation results. In addition, the molecular weight was differentiated (<3 kDa, 3-10 kDa, >10 kDa), and the inhibitory activity was best at >10 kDa. The effect of regulating blood sugar was confirmed by cell experiments. We found that the concentration above 3000 μg/ml groups showed cytotoxicity. Therefore, the subsequent cell experiments were only conducted for groups below 2000 μg/ml.The results showed that the 2000 μg/ ml can inhibit about 30% DPP-4 activity and had dose-dependent manner. In addition, the algae residue added ethanol to extract polyphenols. The yield of polyphenols per gram of dry algae residues was 215 mg. It showed that the polyphenols extracted from ethanol are used as suitable extraction conditions. Based on the above results, the algae active peptide has the potential to be developed into blood pressure and blood sugar health care products, and the effect of polyphenols can be explored.