title: Site-specific, covalent immobilization of BirA by microbial transglutaminase: A reusable biocatalyst for in vitro biotinylation.
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.
authors.affiliation: School of Pharmacy, Weifang Medical University, 261053 Weifang, China.
authors.forename: Chang-Mei
authors.initials: CM
authors.lastname: Yu
authors.affiliation: School of Pharmacy, Weifang Medical University, 261053 Weifang, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhou
authors.affiliation: School of Pharmacy, Weifang Medical University, 261053 Weifang, China.
authors.forename: Wei-Fen
authors.initials: WF
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, Weifang Medical University, 261053 Weifang, China. Electronic address: yanghongming2006@sohu.com.
authors.forename: Hong-Ming
authors.initials: HM
authors.lastname: Yang
authors.affiliation: School of Pharmacy, Weifang Medical University, 261053 Weifang, China. Electronic address: tangjb@wfmc.edu.cn.
authors.forename: Jin-Bao
authors.initials: JB
authors.lastname: Tang
mesh_terms: D019921:Biotinylation; D019731:Carbon-Nitrogen Ligases / Q000737:chemistry*; D002384:Catalysis; D004800:Enzymes, Immobilized / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D060526:Magnetic Fields; D008863:Microspheres; D012097:Repressor Proteins / Q000737:chemistry*; D011503:Transglutaminases / Q000737:chemistry*
chemical_list: D004800:Enzymes, Immobilized; D029968:Escherichia coli Proteins; D012097:Repressor Proteins; D011503:Transglutaminases; D019731:Carbon-Nitrogen Ligases; C058125:birA protein, E coli
keywords: BirA; Magnetic microspheres; Microbial transglutaminase; Site-specific and covalent immobilization