One step purification-immobilization of fuculose-1-phosphate aldolase, a class II DHAP dependent aldolase, by using metal-chelate supports

被引：22

作者：

Ardao, Ines ^{[1
]}

Benaiges, M. Dolors ^{[1
]}

Caminal, Gloria ^{[1
]}

Alvaro, Gregorio ^{[1
]}

机构：

[1] Univ Autonoma Barcelona, Dept Engn Quim, Escola Tecn Super Engn,CSIC, Unitat Biocatalisi Aplicada,IIQA, Bellaterra 08193, Cerdanyola Del, Spain

来源：

ENZYME AND MICROBIAL TECHNOLOGY | 2006年 / 39卷 / 01期

关键词：

fuculose-1-phosphate aldolase (FucA); DHAP dependent aldolase; enzyme immobilization; immobilized metal-chelate affinity chromatography (IMAC); metal-enzyme; one step purification-immobilization of enzymes;

D O I：

10.1016/j.enzmictec.2005.09.001

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

His-tagged recombinant fuculose-1-phosphate aldolase (FucA) from E. coli has been purified by immobilized metal-chelate affinity chromatography (IMAC) at gram scale. During this operation, there was a metal exchange between FucA and the affinity matrix, being the purification yields dependent on the metal nature, which was bound to affinity matrix. One step purification-immobilization of FucA has been carried out on metal-chelate support. The preparation of a FucA immobilized derivative, available to be used as catalyst in aldol addition reactions, has been accomplished in a single step starting from E. coli cell extracts. The best results were obtained with high density support containing Co2+. The immobilization yield was 100% and the immobilized derivative showed 63% of FucA activity initially offered to the support. The best derivative of immobilized FucA is 21-fold more stable than the soluble FucA in DMF/buffer (1:4) at 25 degrees C and it catalyzes aldol addition between S-Cbz-Alaninal and DHAP. (c) 2005 Elsevier Inc. All rights reserved.

引用

页码：22 / 27

页数：6

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