摘要
将聚碳酸乙烯撑酯 (PVCA)与α,ω-双端氨基聚乙二醇 (H2 N-PEG-NH2 )溶于 DMF,于液蜡中进行交联反应制得亲水性固定化酶载体 ,将其与胰蛋白酶进行偶联反应制备了固定化胰蛋白酶 .酶蛋白的比活力及其于载体上的结合量与反应条件有关 ,当 w(PVCA) / w(H2 N-PEG-NH2 )为 0 .5时 ,二者均处于最高值 .此固定化酶酶促反应的最适 p H值和 Km 值均较之溶液酶有显著提高 。
In this paper, poly(ethylene oxide) end capped with amine groups(PEO) was prepared and dissolved with poly(vinylene carbonate)(PVCA) in DMF solution. The solution was suspended in paraffin oil to carry out the crosslinking reaction and produce beadlike hydrophilic supports containing reactive cyclic carbonate groups for enzyme immobilization. In above reaction, PEO was acted as both crosslinking agent and hydrophilic modifier of poly(vinylene carbonate). Evaluation of the supports using trypsin as a model enzyme shows that the specific activity of immobilized trypsin and amount of enzymes coupled to supports were related to PVCA/PEO ratio, reaction time, concentration and pH value of enzyme solution, etc.. The amount of trypsin coupled and the specific activity reach their maximum at w (PVCA)/ w (PEG)= 1/2 . The optimal pH value for trypsin immobilized increases as compared with the optimal pH value for native trypsin. The K m value of immobilized and native trypsin was determined to be 32 5 mmol/L and 15 2 mmol/L, respectively.
出处
《高等学校化学学报》
SCIE
EI
CAS
CSCD
北大核心
2001年第4期678-682,共5页
Chemical Journal of Chinese Universities
基金
国家自然科学基金! (批准号 :5 9973 0 0 9)资助
