• 作者： 簡靜香; 張勝祺; 魏耀揮
• 作者服務機構： 國立陽明醫學院生化學科
• 中文摘要： 神經胺酸??（neuraminidase, E.C. 3.2.1. 18）可由clostridium perfringens ATCC 10543菌株之培養液中純化而得
  。純化的步驟包括硫銨沈澱，膠體過濾層析，陰離子交換層析和等電點焦集快速蛋白液相層析等方法，所得的均質狀態
  酵素比活性為289單位／毫克蛋白，收率為25%。
  此酵素由單一之多胜?鏈組成，經膠體管柱層析及聚丙烯醯胺膠體電泳測定之分子量為69,000；以涎酸?乳糖
  （sialyllactose）為酵素基質，所得之Km為1.5 mM, Vmax 為0.41 μmoles/min/ml（酵素濃度為0.14 μg/ml）。此酵
  素之反應最適pH為6.0，在pH 5.2至8.0範圍，亦相當穩定。純化後的酵素濃溶液在4℃保存一年，其活性並無明顯
  變化。
  純化之酵素可完全水解人類血液中之酸糖蛋白（α1-acid glycoprotein），故可被用以檢驗血清中神經胺酸（neura-
  minic acid）含量之醫檢酵素。m
• 英文摘要： Neuraminidase (EC 3.2.1.18) has been purified from the culture medium of Clostridium perfringensATCC 10543, through steps of gel filtration on Sephadex G-75 column, DEAE-cellulose DE 23 anionexchange chromatography, and isochromatofocusing. A homogeneous enzyme was obtained with a 7552-fold increase in specific activity to 295 units/mg protein. The yield was about 25%. The enzyme consists of a single polypeptide with a molecular weight of 69,000 as determined bySDS-polyacrylamide gel electrophoresis. Kinetic studies showed that Km is 1.5 mM for sialyllactose andVmax is 0.41 μmole/min/ml at the enzyme concentration of 0.14 μg/ml. The enzyme is stable at pH 5.2-8.0 with an optimal pH of 6.0. A concentrated solution of the purified enzyme was stable over one year at4℃. The purified enzyme hydrolyzed human α1-acid glycoprotein completely; thus, it can be used in theclinical assay of N-acetylneuraminic acid in the serum.
• 中文關鍵字： neuraminidas; Clostridium perfringens
• 英文關鍵字： --