新型交联剂三羟甲基磷固定化脂肪酶的研究

Lipase Immobilization with a New Crosslinking Reagent Tris(hydroxymethyl)phosphine

摘要:

本试验研究了一种新型交联剂三羟甲基磷（THP）用于以纯棉浴巾载体、聚乙烯亚胺（PEI）为吸附剂的固定化脂肪酶制备新方法。通过单因子实验获得了固定化条件为：0.2 g浴巾使用0.5 mL PEI（浓度3.37 mg/mL，pH 7.0）、2 mL THP（浓度为0.10%，V/V）、固定化温度为40 ℃、交联时间为20 min。制备的固定化脂肪酶催化正丁酸乙烯酯与戊醇酯交换反应，1 h底物转化率达92.1%，酶比活力达到15.80 U/g固定化酶。与戊二醛（GA）交联法制备的固定化酶相比，本固定化酶具有良好的重复使用稳定性、温度稳定性和高比活力，本试验采用的方法是一种低成本、实用性强的脂肪酶固定化新方法。

Abstract:

In the present work, tris(hydroxymethyl)phosphine (THP) was used as a new crosslinking reagent in lipase immobilization on cotton towel with adsorbent polyethylenimine (PEI). The optimal conditions of immobilization by single factors test were obtained as follows: 0.5 mL of PEI (3.37 mg/mL, pH 7.0) for each 0.2 g cotton towe, 2ml of THP [0.10% (V/V)], immobilization temperature 40 ℃ and crosslinking reaction time 20 min. THP-immobilized lipase can catalyze interesterification of vinyl butyric acid and pentanol. The conversion rate in 1h achieved 92.1%, and the activity of immobilized lipase was 15.80 U/g. Compared with the GA-immobilized lipase, THP-immobilized lipase had higher reusability, thermal stability and activity, indicating this is a low cost and practical newly method for lipase immobilization.