Chia Nan University of Pharmacy & Science Institutional Repository:Item 310902800/25048
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    標題: 鯖魚內臟蛋白酶之性質
    Purification and Properties of Mackerel Visceral Proteinases
    作者: 郭建民
    孫寶年
    貢獻者: 嘉南藥專食品衛生科
    國立海洋學院水產食品科學研究所
    關鍵字: Mackerel
    Scomber australasicus
    Proteinase
    Purification
    日期: 1986
    上傳時間: 2012-02-24 16:02:20 (UTC+8)
    摘要: 新鮮鯖魚(Scomber australasicus)內臟經酒精處理及水洗後,製成丙酮粉,再以去離子水由丙酮粉抽取酵素,然後經30-70%硫銨分劃,Ultrogel AcA 44膠濾及DEAE-Sephadex A-50管柱層析後得兩種蛋白酶,分別稱為A及B,在不連續膠體電泳上A僅呈一絛電泳帶(band),為純化之蛋白酶, B為兩條電泳帶。依照基質特異性推測,A及B皆為胰蛋白酶。胰蛋白酶A及B水解TAME的最適pH分別在9.0及7.6左右,水解酪蛋白最適溫度分別在45℃及55℃。A及B在50℃以下皆安定,起過80℃均失活。在中性及微鹼性都安定。以SDS-polyacrylamide膠體電泳測定, A分子量為29,200。蛋白酶A稍受金屬離于Na+、Co2+之活化及受 Hg2+之抑制,蛋白酶B稍受Na+ 、EDTA活化及受Ca2+、Hg2+之抑制。以TAME為基質,A之Km為9.1 x l0-4 M, Vmax為5.6 x l0 5units/mole enzyme/min。
    Proteinases in acetone powder of mackerel viscera were extracted with deion-ized water and purified by 30-70% ammonium sulfate fractionation, Ultrogel AcA. 44 gel fltration, and DEAE-Sephadex A-50 ion exchange chromatography.Two proteinase peaks having caseinolytic activity, named A and B respectively,were obtained from the DEAE-Sephadex A-50 elution. Proteinase A was homogenous showing a single band on disc-polyacrylamide gel electrophoresis, while proteinase B yieded two bands. Both proteinase A and B were trypsin-like enzymes on the basis of authentic substrate specificity.The optimum pH of mackerel visceral proteinase A and B for hydrolysis of TAME was found to be 9.0 and 7.6 respectively. The optimum temperature of A and B for hydrolysis of casien was 45℃ and 55℃ respectively. Both proteinases were stable below 50℃ in neutral and slightly alkaline pH. Activity disappeared above 80℃, or at acidic pH. Molecular weight of the proteinase A was determined to be 29,200 by SDS-polyacrylamide gel electroohoresis. The activity of proteinase A was slightly activated by Na+ and Co2+, and was inhibited by Hg2+. Proteinase B was slightly activited by Na+ and EDTA, and was inhibited by Ca2+ and Hg2+. The Michaelis constant (Km), and maximal velocity (Vmax) of proteinase A were found to be 9.1 x 10-4M and 5.6 x 10 5 unit/mole enzyme/min respectively
    關聯: 嘉南學報 12:p.A1-A9 轉載 食品科學 11(3-4) : p.189-197 (1984)
    显示于类别:[嘉南學報] 12期 (1986)
    [ 食品科技系 ] 期刊論文

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