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Site-specific, covalent immobilization of BirA by microbial transglutaminase: A reusable biocatalyst for in vitro biotinylation

Author:
Yu, Chang-Mei, Zhou, Hui, Zhang, Wei-Fen, Yang, Hong-Ming, Tang, Jin-Bao
Source:
Analytical biochemistry 2016 v.511 pp. 10-12
ISSN:
0003-2697
Subject:
Escherichia coli, biocatalysts, biotin, biotinylation, crosslinking, enzyme activity, immobilized enzymes, magnetic separation, magnetism, microparticles, protein-glutamine gamma-glutamyltransferase, recycling
Abstract:
A facile approach for the production of a reusable immobilized recombinant Escherichia coli biotin ligase (BirA) onto amine-modified magnetic microspheres (MMS) via covalent cross-linking catalyzed using microbial transglutaminase (MTG) was proposed in this study. The site-specifically immobilized BirA exhibited approximately 95% of enzymatic activity of the free BirA, and without a significant loss in intrinsic activity after 10 rounds of recycling (P > 0.05). In addition, the immobilized BirA can be easily recovered from the solution via a simple magnetic separation. Thus, the immobilized BirA may be of general use for in vitro biotinylation in an efficient and economical manner.
Agid:
6056304