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来自不定毛壳菌的葡聚糖酶的固定化。

Immobilization of dextranase from Chaetomium erraticum.

作者信息

Erhardt Frank Alwin, Jördening Hans-Joachim

机构信息

Institute for Technical Chemistry, Department for Carbohydrate Technology, Technical University Braunschweig, Hans-Sommer-Strasse 10, D-38106, Braunschweig, Germany.

出版信息

J Biotechnol. 2007 Sep 30;131(4):440-7. doi: 10.1016/j.jbiotec.2007.07.946. Epub 2007 Jul 31.

Abstract

In order to facilitate the Co-Immobilization of dextransucrase and dextranase, various techniques for the immobilization of industrial endo-dextranase from Chaetomium erraticum (Novozymes A/S) were researched. Adsorption isotherms at various pH-values have been determined for bentonite (Montmorillonite), hydroxyapatite and Streamline DEAE. Using bentonite and hydroxyapatite, highest activity loads (12,000 Ug(-1); 2900 Ug(-1), respectively) can be achieved without a significant change of the apparent Michaelis-Menten constant K(M). For successful adsorption, enzyme to bentonite ratios greater than 0.4 (w/w) have to be used as lower ratios lead to 90% enzyme inactivation due to bentonite contact. In addition, covalent linkage using the activated oxiran carriers Eupergit C and Eupergit C250L as well as linkage with aminopropyl silica via metaperiodate activation of glycosyl moiety of dextranase are discussed. This is also the first report probing the structure of a matrix containing dextranase by use of substrate species with different molecular weights. From this we can observe a relationship between the porosity of Eupergit and dextran dependent activity. For the reactor concept using Co-Immobilisates, hydroxyapatite will be preferred to Eupergit because of its higher specific activity and dispersity.

摘要

为了促进葡聚糖蔗糖酶和葡聚糖酶的共固定化,研究了多种固定化来自多变毛壳菌(诺维信公司)的工业内切葡聚糖酶的技术。已测定了膨润土(蒙脱石)、羟基磷灰石和Streamline DEAE在不同pH值下的吸附等温线。使用膨润土和羟基磷灰石时,在表观米氏常数K(M)无显著变化的情况下,可分别实现最高活性负载(分别为12,000 Ug(-1);2900 Ug(-1))。为了成功吸附,酶与膨润土的比例必须大于0.4(w/w),因为较低的比例会因与膨润土接触导致90%的酶失活。此外,还讨论了使用环氧乙烷活化载体Eupergit C和Eupergit C250L的共价连接以及通过高碘酸盐活化葡聚糖酶糖基部分与氨丙基硅胶的连接。这也是首次通过使用不同分子量的底物种类来探究含有葡聚糖酶的基质结构的报告。由此我们可以观察到Eupergit的孔隙率与葡聚糖依赖性活性之间的关系。对于使用共固定化产物的反应器概念,由于其较高的比活性和分散性,羟基磷灰石将比Eupergit更受青睐。

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