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標題: | 虱目魚鱗胜肽衍生物之生物功能性分析 Biological function analysis of the peptides derivatives from milkfish(Chanos chanos) scales |
作者: | Chun-Hui Huang 黃均蕙 |
指導教授: | 李英周(Yin-Chou Lee) |
關鍵字: | 抗氧化活性,金屬螯合活性,最小抑菌濃度,木瓜蛋白酵素, Antioxidant activity,Iron chelating activity,Minimum inhibitory concentration,Papain, |
出版年 : | 2017 |
學位: | 碩士 |
摘要: | 魚鱗等副產物僅少數能再加工創造二次經濟價值,早期作為廢棄物處置,造成環境污染和社會成本增加,若能回收廢棄魚鱗中的膠原蛋白作為生物活性胜肽原料,不僅能轉變為具附加價值的產品同時降低資源的浪費。魚鱗膠原蛋白的水解胜肽被發現具有生物功能如抗氧化功能、抗菌功能、降血壓、抑制腫瘤等功能,且因界面活性表面應可用於食品乳化劑、抑制冰晶形成作為蛋白質保護劑、抑菌功能在農業上則可降低草莓灰黴病發病率。本研究使用虱目魚魚鱗以木瓜酵素水解成胜肽後進行膠原蛋白含量測試、胺基酸組成分析,同時依分子量分為大分子量 (>10kDa)、中分子量 (3-10kDa)及小分子量胜肽(<3kDa)。小分子量胜肽進行分子量測試及定序,再利用Mobyle @ RPBS Net進行其結構預測。將大中小分子量胜肽進行功能測試分別為抗氧化能力、金屬離子螯合能力、還原力、抑菌率及最小抑菌濃度測試。結果指出,虱目魚鱗的膠原蛋白含量為12%,其中 Gly、Pro、Hyp 胺基酸含量最高,分別為27%、11%、10.4%。小分子胜肽分子量介於567-2775 Da間,其中胜肽2693含27個胺基酸,序列為KPTKDGVTVAVISGEALGIKICFTSTG,有40%的疏水性胺基酸且帶電量為+1,結構為β-sheet,構型特殊因此委託明欣生技公司合成。虱目魚魚鱗胜肽、>10kDa、3-10kDa、<3kDa及胜肽2693抗氧化能力(DPPH自由基清除能力,濃度1μg/ml)分別為51.27%、41.28%、63.98%、64.20%及52.01%,亞鐵離子螯合能力(freeozine model,EDTA濃度1μg/ml) 分別為40.93%、75.99%、68.09%、5.76%但胜肽2693無亞鐵離子螯合能力;還原力結果(FRAP,EDTA1 μg/ml)分別為10.26%、17.18%、14.11%、8.66%及3.06%, <3kDa及2693 peptide對金黃色葡萄球菌(Staphylococcus aureus)最小抑菌濃度(MIC)結果為7μg/ml (83.3%)及15-30μg/ml (84.3%),其餘無抑菌能力。虱目魚鱗膠原蛋白胜肽含多種生物活性胜證實能成功的將水產品廢棄物有效利用,未來可發展醫療、食品等產業,則可針對胜肽的功能性來選擇蛋白酵素分離不同分子量的胜肽。 The hydrolyzed peptides of fish scale collagen are founded to have biological function such as antioxidant activity, antibacterial activity, lowering blood pressure, inhibiting tumor and so on… In this study, milkfish scales are used. The collagen content of milkfish scale are analyzed by soften the fish scale and hydrolyze the fish scale into collagen peptide. The compositions of amino acid are divided into three groups based on the molecular weight: high molecular peptides (>10kDa), medium molecular weight peptides (3-10kDa) and small molecular weight peptides (<3kDa). The molecular weight distribution of MALDI-TOF / MS is measured by small molecular weight peptides and its structure is predicted by Mobyle @ RPBS Net. The result shows that the content of collagen was 12%, and the contents of Gly, Pro and Hyp were 27%, 11% and 10.4% respectively. The molecular weight of the small molecule peptide is between 567-2775, in which the peptide 2693 contains 27 amino acids and the sequence is KPTKDGVTVAVISGEALGIKICFTSTG. There is 40% hydrophobic amino acid and the charge is +1 and the structure is β-sheet. The antioxidant capacity of milkfish peptide, >10kDa, 3-10kDa, <3kDa and peptide 2693 were 51.27%, 41.28%, 63.98%, 64.20% and 52.01%, respectively. The ferrous ion chelating ability was 40.93%, 75.99%, 68.09%, 5.76%, respectively, while peptide 2693 doesn’t have such ability. And the result of recovering is 10.26%, 17.18%, 14.11%, 8.66% and 3.06%, respectively. As for <3kDa and 2693 peptide, the minimum inhibitory concentration of Staphylococcus aureus was 7μg / ml (83.3%) and 15-30μg / ml (84.3%). The milkfish peptide produces biologically active peptides by different proteases. The differences of these biologically active peptides are depending on the different molecular weight, amino acid composition and stereospecific structure. The more hydrophobic amino acids and intramolecular hydrogen bonds are more possible to enhance antioxidant activity. The larger space of three-dimensional structure of peptide ensures more metal ions. There is more than 40% hydrophobic amino acid and positive charage in <3kDa small molecules. According to the simulation interaction between peptides 2693 and bacterial cell, we presume that the bacteria are the barrel-attack type of cell membrane. Milkfish scale peptide containing several of biological activities are verifying the effective using of aquatic waste. It is possible that fish scale peptide are using in medical, food and other industries in the future development. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/20617 |
DOI: | 10.6342/NTU201702497 |
全文授權: | 未授權 |
顯示於系所單位: | 漁業科學研究所 |
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