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

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• 作者：Chang-Mei Yu ; Hui Zhou ; Wei-Fen Zhang ; Hong-Ming Yang ; ^{yanghongming2006@sohu.com" class="auth_mail" title="E-mail the corresponding author} ; Jin-Bao Tang ; ^{tangjb@wfmc.edu.cn" class="auth_mail" title="E-mail the corresponding author}
• 关键词：BirA ; Site-specific and covalent immobilization ; Magnetic microspheres ; Microbial transglutaminase
• 刊名：Analytical Biochemistry
• 出版年：2016
• 出版时间：15 October 2016
• 年：2016
• 卷：511
• 期：Complete
• 页码：10-12
• 全文大小：562 K

文摘

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.