通过离子吸附将糖化酶可逆固定在涂有聚乙烯亚胺的Sepabeads上。

Reversible immobilization of glucoamylase by ionic adsorption on sepabeads coated with polyethyleneimine.

作者信息

Torres Rodrigo, Pessela Benevides C C, Mateo Cesar, Ortiz Claudia, Fuentes Manuel, Guisan Jose M, Fernandez-Lafuente Roberto

机构信息

Departamento de Biocatálisis, Instituto de Catálisis, CSIC, Campus Universidad Autónoma, Cantoblanco, 28049 Madrid, Spain.

出版信息

Biotechnol Prog. 2004 Jul-Aug;20(4):1297-300. doi: 10.1021/bp049943g.

DOI:10.1021/bp049943g

PMID:15296467

Abstract

Glucoamylase (GA) from Aspergillus niger was immobilized via ionic adsorption onto DEAE-agarose, Q1A-Sepabeads, and Sepabeads EC-EP3 supports coated with polyethyleneimine (PEI). After optimization of the immobilization conditions (pH, polymer size), it was observed that the adsorption strength was much higher in PEI-Sepabeads than in Q1A-Sepabeads or DEAE-supports, requiring very high ionic strength to remove glucoamylase from the PEI-supports (e.g., 1 M NaCl at pH 5.5). Thermal stability and optimal temperature was marginally improved by this immobilization. Recovered activity depended on the substrate used, maltose or starch, except when very low loading was used. The optimization of the loading allowed the preparation of derivatives with 750 IU/g in the hydrolysis of starch, preserving a high percentage of immobilized activity (around 50%).

摘要

来自黑曲霉的葡糖淀粉酶（GA）通过离子吸附固定在涂有聚乙烯亚胺（PEI）的DEAE - 琼脂糖、Q1A - Sepabeads和Sepabeads EC - EP3载体上。在优化固定条件（pH、聚合物尺寸）后，观察到PEI - Sepabeads上的吸附强度远高于Q1A - Sepabeads或DEAE载体，从PEI载体上去除葡糖淀粉酶需要非常高的离子强度（例如，在pH 5.5时为1 M NaCl）。这种固定化略微提高了热稳定性和最适温度。回收的活性取决于所用的底物，麦芽糖或淀粉，除非使用非常低的负载量。负载量的优化使得能够制备在淀粉水解中具有750 IU/g的衍生物，同时保留高比例的固定化活性（约50%）。