共价固定化灰黄青霉α-半乳糖苷酶及其在低聚糖水解中的应用。

Covalent immobilization of alpha-galactosidase from Penicillium griseoroseum and its application in oligosaccharides hydrolysis.

机构信息

Departamento de Bioquimica e Biologia Molecular-BIOAGRO, Universidade Federal de Viçosa, Viçosa 36570.000 Minas Gerais, Brazil.

出版信息

Appl Biochem Biotechnol. 2009 Sep;158(3):540-51. doi: 10.1007/s12010-008-8387-9. Epub 2008 Oct 21.

DOI:10.1007/s12010-008-8387-9

PMID:18937072

Abstract

Partially purified alpha-Galactosidase from Penicillium griseoroseum was immobilized onto modified silica using glutaraldehyde linkages. The effective activity of immobilized enzyme was 33%. Free and immobilized alpha-galactosidase showed optimal activity at 45 degrees C and pH values of 5 and 4, respectively. Immobilized alpha-galactosidase was more stable at higher temperatures and pH values. Immobilized alpha-galactosidase from P. griseoroseum maintained 100% activity after 24 h of incubation at 40 degrees C, while free enzyme showed only 32% activity under the same incubation conditions. Defatted soybean flour was treated with free and immobilized alpha-galactosidase in batch reactors. After 8 h of incubation, stachyose was completely hydrolyzed in both treatments. After 8 h of incubation, 39% and 70% of raffinose was hydrolyzed with free and immobilized alpha-galactosidase respectively. Immobilized alpha-galactosidase was reutilized eight times without any decrease in its activity.

摘要

从玫瑰青霉中部分纯化的α-半乳糖苷酶通过戊二醛键连接固定在改性硅胶上。固定化酶的有效活性为 33%。游离和固定化的α-半乳糖苷酶的最适活性分别在 45°C 和 pH 值为 5 和 4 时达到。固定化的α-半乳糖苷酶在较高的温度和 pH 值下更稳定。固定化的α-半乳糖苷酶从玫瑰青霉在 40°C 孵育 24 小时后保持 100%的活性，而游离酶在相同的孵育条件下仅显示 32%的活性。脱脂大豆粉在分批反应器中用游离和固定化的α-半乳糖苷酶处理。孵育 8 小时后，两种处理方式均完全水解棉子糖。孵育 8 小时后，游离和固定化的α-半乳糖苷酶分别水解棉子糖 39%和 70%。固定化的α-半乳糖苷酶重复使用了八次，其活性没有任何下降。