利用連續式酵素膜反應器水解豬血生產含生物活性胜肽之保健產品

Bioactive peptides production by hydrolysis of porcine blood proteins in a continuous enzymatic membrane reactor

魏任廷(Jen-Ting Wei)

指導教授 ： 蔣丙煌

國立臺灣大學/生物資源暨農學院/食品科技研究所/博士(2009年)

https://doi.org/10.6342/NTU.2009.10582

摘要

豬隻全血液中約有17 ~ 18 %之粗蛋白質，為一良好的動物性蛋白質來源。本研究分別以豬血球、血漿、去纖維蛋白原血漿為受質，於幾種酵素組合下進行批式水解，探討生產生物活性胜肽的可行性。本研究發現，以血球為受質，添加複合酵素(trypsin、chymotrypsin、thermolysin)進行水解，水解6小時，可得到較佳抑制ACE能力之水解物，IC50為0.58 mg/mL；水解10小時可得到較佳的清除DPPH自由基，清除能力為65%，而且將該水解液添加在以β-amyloid(25-35)處理之RBE4細胞中，RBE4細胞存活率可由67%升高至82%，因此具潛在預防Alzheimer's Disease效果。血球之酵素水解液亦具有其他機能性，包括還原力、促進皮膚纖維母細胞增生、免疫調節、抑制腫瘤細胞生長等，雖然所表現之效果未優於其他已知產品或研究，但在一水解產品中同時存在各類機能性亦屬難得。
　　由於豬血球水解液具清除DPPH自由基，且同時在抑制ACE能力及存在其他功能性，值得發展成產品。為有效率生產含生物活性胜肽且具生理機能性之產品，本研究進一步利用膜反應器系統，探討連續式量產之可行性。在膜反應器中，當酵素與受質比為1:5，滯留時間為100分鐘時，可在2小時達到穩定之操作狀態。在穏定狀態下，產品之ACE抑制率可達86%，清除DPPH自由基能力為54%。同時，膜反應器生產之產品為黃色透明，血色素已被去除。
　　本研究證實豬血水解物具有潛在抗氧化、降血壓及存在其他功能性，且在膜反應器中利用混合酵素水解豬血球，可連續式水解生產生物活性胜肽，所以，利用膜反應器酵素水解是一具潛力之生產含生物活性胜肽且具生理機能性產品加工方法。

關鍵字

豬血 ； 酵素水解 ； 生物活性胜肽膜反應器

並列摘要

Porcine blood with 17~18% protein content is a good source of protein for food. In this study, hydrolysates of red blood corpuscle, blood plasma and defibrinated blood plasma were prepared by enzymatic hydrolysis with several proteases, and evaluated for the possibility of using the enzymatic processes for manufacturing bioactive peptides. It was found that the hydrolysate produced by hydrolyzing red blood corpuscle with the mixture of trypsin, chymotrypsin and thermolysin had the highest ACE inhibitory activity (IC50 = 0.58 mg/mL) and the highest scavenging effect on DPPH (scavenging effect = 65%) after 6 h and 10 h hydrolysis, respectively. For the potential protection against Alzheimer's disease, addition of the hydrolysate into the β-amyloid(25-35) treated RBE4 cell culture could raise the survival rate of RBE4 cell from 67% to 82%. In respect of other functionalities, including reducing power, proliferation effect on the skin fibroblast WS1 cell, immunomodulatory effect, anti-proliferative activity on the tumor cell line, although the hydrolysates did not show superior capabilities than the other well-known products, this study proved that the hydrolysate did possess such functionalities mentioned above.
　　Because of the health-promoting functions possessed by the hydrolysate of porcine red blood corpuscles, attempt was made to evaluate the possibility of manufacturing the product continuously using enzymatic membrane reactor. It was found that when the hydrolysis was carried out with an enzyme/substrate ratio of 1:5 and a residence time of 100 min, the process reached steady state in 2 h. The ACE-inhibitory activity of the product during the steady state process was 86% and its scavenging effect on DPPH was 54%. The membrane process also decolourised the enzyme-hydrolysed product, thus improving the appearance of the product.
　　This study demonstrated that hydrolysates of porcine blood possess antihypertensive and antioxidant activities. Using red blood corpuscles as the substrate, the hydrolysis could be carried out in a membrane reactor with a mixture of proteases to produce bioactive peptides continuously. Therefore processing of porcine blood in an enzymatic membrane reactor is a potential method for producing a health-promoting product.

並列關鍵字

porcine blood ； enzymatic hydrolysis ； bioactive peptides ； membrane reactor

參考文獻

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