β-D-半乳糖苷酶壳聚糖固定方法、特性和应用研究.docVIP

β-D-半乳糖苷酶壳聚糖固定方法研究β-D-半乳糖苷酶水解奶中的乳糖是解决“乳糖不耐症”的有效方法。本研究以壳聚糖为载体和戊二醛为交联剂，采用共价交联和吸附交联的方法，优选出不同形态壳聚糖固定化乳糖酶的方法，并进一步研究了固定化乳糖酶的特性和应用的效果。另外，还比较研究了测定乳糖水解率的方法，优选出了简便、准确和快捷的测定方法。主要得出以下结论： 1. 对碘量法℃下固定12h。 4. 壳聚糖膜与戊二醛共价交联法固定乳糖酶的特性发生了变化与游离酶相比℃升至45℃, 60℃保温1h后仍保留初始活力的50%，而游离酶则几乎完全失活；米氏常数Km固定化酶重复使用8次后其活力仅损失10%；4～12℃下25d后固定化酶活力仍保留50%以上。  STUDY ON METHODS, CHARACTERS AND APPLICATIONS OF CHITOSAN IMMOBILIZEDβ-D-GALACTOSIDASE ABSTRACT “Lactose malabsorption” is the bottleneck of the dairy industry development in our country. To make use of immobilizedβ-D-galactosidase to hydrolyze lactose in milk can settle “lactose malabsorption” effectively.This study adopted chitosan as carrier, and glutaraldehyde as crosslinked reagent to immobilizeβ-D-galactosidase. We used ways of covalent-crosslinked and absorption- crosslinked to find the conditions to immobilize lactase in different forms of chitosan in order to pick out an optimal condition for immobilization. Based on above researches, we had studied the characteristics of the immobilized lactase that had been prepared under the optimal conditions. Additional, we had compared five methods which mensurated ratio of hydrolyzed lactose in order to find a method that is simple、nicety and economy. The results were as follows: 1. Compared among five methods including Fehling method, the colorimetric method of Anthrone, the Sacchairmetry, iodine number method and enzyme catalyzed method. The result showed that enzyme catalyzed method was a nicety, economy, simple and convenient way to mensurate content of lactose. 2. Studied of the methods of three forms of chitosan including membrane、gelatin and bead to immobilizeβ-D-galactosidase as well as activity recycle rate. The results showed that the activity recycle rate of chitosan bead adsorption-crosslinked with glutaraldehyde immobilized lactase reached to 52.9%, chitosan gel covalent-crosslinked with glutaraldehyde reached immobilized lactase to 71.1%, chitosan membrane covalent-crosslinked with gluta