Inhibition on Angiotensin I Converting Enzyme from Shellfish Muscle Protein Hydrolysates (I)

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Project title

Code/計畫編號

NSC92-2321-B019-003

Translated Name/計畫中文名

台灣養殖貝類生理活性物質的分離鑑定與功能評估(一)蜆(二)文蛤(三)牡蠣---貝類肌肉蛋白質水解物對血管升壓素轉換脢之抑制(I)

Project Coordinator/計畫主持人

Jenn-Shou Tsai

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Food Science

Website

https://www.grb.gov.tw/search/planDetail?id=923379

Year

2003

Start date/計畫起

01-08-2003

Expected Completion/計畫迄

01-07-2004

Bugetid/研究經費

750千元

ResearchField/研究領域

食品科技(農)
漁業

以本省花蓮產之黃金蜆為原料，將生鮮蜆肉以熱水抽出法或將剩餘蜆肉經由凍乾成粉末後，利用蛋白酶 Protease N(PN)、Protamex(PX)於 50℃進行一次或二次 (Flavourzyme)水解，探討熱水抽出物與蜆肉粉末水解物對血管升壓素轉換酶(ACE) 之抑制及降高血壓的生理效果。由熱水抽出法得到的抽出物中可溶性蛋白質與胜含量分別為 84 mg/g 與 85 mg/g，而對 ACE 之抑制 IC50 值為 1.95 mg/ml。蜆肉粉末水解物中可溶性蛋白含量，以 Protease N 一次水解 2 小時者(PN2)最高，為 635 mg/g。胜含量以 Protease N 一次水解五小時再經 Flavourzyme 二次水解 0.5 小時者(PN5F0.5)最高，為 348 mg/g。對 ACE 抑制能力而言，以 Protamex 一次水解 5 小時(PX5)者最佳，IC50 值為 0.043 mg/ml。蜆肉粉末水解物以 Protamex 一次水解 2 小時再經 Flavourzyme 二次水解 0.5 小時(PX2F0.5)之產物不具苦味。另一方面，PX5 水解物對 ACE 的抑制屬於混合型，其 Ki 值為 0.032 mg/ml。將此水解物以膠體層析法進行劃分，可得到四個主要波峯，其中以分子量介於 384 至 325 Da 間之劃分物具有最強的 ACE 抑制活性。The muscle of golden freshwater clam (Corbicula fluminea, Muller)(cultivated in Hualien County) was extracted using hot water and the freezing dried residual muscle was hydrolyzed by Protease N (PN) and Protamex (PX) as primary hydrolysis followed by a secondary hydrolysis (Flavourzyme, FL) at 50℃. The effects of hot water extracts and hydrolysates on inhibitory activity against angiotensin I converting enzyme (ACE) was investigated. The soluble protein and peptide content of hot water extracts were 85 mg/g and 84 mg/g, repectively, and the IC50 of ACE was 1.95 mg/ml. The highest value in soluble protein (635 mg/g) and peptide content (348 mg/g) of hydrolysates were obtained from PN hydrolysis for 2 hr and PN hydrolysis for 5 hr followed by FL hydrolysis for 0.5 hr, respectively. However, the lowest IC 50 on ACE ( 0.043 mg/ml) of hydrolysate was obtained by PX hydrolysis for 5 hr (PX5). The kinetics of the hydrolysate on ACE-inhibition indicated that PX5 and Captopril produced mixed inhibition and competitive inhibition, respectively. Their Ki values were 0.032 mg/ml and 0.0067 μg/ml, respectively. The hydrolysate of PX5 was fractionated by gel permeation chromatography with Sephadex G-25. The fraction D with molecular weight 384-325 Da showed the highest inhibitory efficiency ratio being 1314.7 %/mg/ml.

Keyword(s)

蜆
熱水抽出物
蛋白質水解物
血管升壓素轉換酶(ACE)
降高血壓
Freshwater clam
Protein hydrolysate
Angiotensin I converting enzyme

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Inhibition on Angiotensin I Converting Enzyme from Shellfish Muscle Protein Hydrolysates (I)

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