Abstract
An amino-modified glass surface for enzymatic protein immobilization by microbial transglutaminase (MTG) was developed. Diamine substrates with secondary amino groups in the linker moiety, like triethylenetetramine (TETA), exhibited at most a 2-fold higher reactivity in the MTG-catalyzed reaction compared to those with the alkyl linker. A 96-well glass plate was subsequently modified with selected diamine substrates. Validation of the modified surface by enzymatic immobilization of enhanced green fluorescent protein tagged with a glutamine donor-substrate peptide (LLQG) of MTG revealed that the protein loading onto the TETA-modified glass surface was approximately 15-fold higher than that on the unmodified one.
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Acknowledgments
We are grateful to Ajinomoto Co., Inc. for providing the sample of MTG. The present work was supported by the Industrial Technology Research Grant Program in 2006 from the New Energy and Industrial Technology Development Organization (NEDO) of Japan (to N.K.).
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Tanaka, Y., Doi, S., Kamiya, N. et al. A chemically modified glass surface that facilitates transglutaminase-mediated protein immobilization. Biotechnol Lett 30, 1025–1029 (2008). https://doi.org/10.1007/s10529-008-9656-y
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DOI: https://doi.org/10.1007/s10529-008-9656-y